Background
==========

Polysaccharide-rich fungi and plants have been employed for centuries by cultures around the world for their dietary and medicinal benefits \[[@B1]-[@B5]\]. Often thought to merely support normal bowel function and blood glucose and lipid levels \[[@B6]-[@B8]\], certain polysaccharides have attracted growing scientific interest for their ability to exert marked effects on immune system function, inflammation and cancers \[[@B9]-[@B11]\]. Many of these chemically and structurally diverse, non- to poorly-digestible polysaccharides have been shown to beneficially affect one or more targeted cellular functions *in vitro*\[[@B11]-[@B16]\], but much of the *in vivo*literature consists of studies in which polysaccharides were injected \[[@B1],[@B2]\]. For clinicians and scientists interested in immunologic effects following dietary intake, the value of such studies is uncertain. Polysaccharides that elicit effects *in vitro*or by injection may be ineffective or have different effects when taken orally \[[@B17]\]. We thus decided to conduct a systematic review to evaluate the specific immunologic effects of dietary polysaccharide products on rodents and human subjects.

Methods
=======

Literature review
-----------------

Studies were identified by conducting electronic searches of PubMed and Google Scholar from their inception to the end of October 2009. The reference lists of the selected articles were checked for additional studies that were not originally found in the search.

Study selection and data extraction
-----------------------------------

The following search terms were combined with the term polysaccharide: dietary AND immune, or oral AND immune, or dietary AND inflammation, or oral AND inflammation. When specific polysaccharides or polysaccharide-rich plants and fungi were identified, further searches were conducted using their names with the same search terms. Studies were selected based on the following inclusion criteria:

1\. Rodent or human studies

2\. The presence of test group and control group (using either placebo, crossover, sham, or normal care)

3\. Studies reporting statistically significant immunomodulatory effects

4\. English language

5\. Studies published up to October 2009.

Two researchers (JER, EDN) reviewed the list of unique articles for studies that fit the inclusion criteria. Uncertainties over study inclusion were discussed between the researchers and resolved through consensus. Searches were then conducted to obtain specific polysaccharide product information: safety (using the search terms: toxicity, NOAEL, LD~50~), composition and structure, and disposition.

Quality assessment
------------------

Each study was assessed as to whether or not it reported a significant outcome measure for the polysaccharide intervention group.

Results
=======

A total of 62 rodent publications (Tables [1](#T1){ref-type="table"}, [2](#T2){ref-type="table"} and [3](#T3){ref-type="table"}) and 15 human publications (Table [4](#T4){ref-type="table"}) were deemed appropriate for inclusion in this review. Available structural and compositional information for these immunomodulatory polysaccharides are provided in Table [5](#T5){ref-type="table"} and safety information is provided in Table [6](#T6){ref-type="table"}. The majority of animal studies explored models in which animals were injected or implanted with cancer cells or tumors, were healthy, or were exposed to carcinogens. Other studies investigated immunodeficient, exercise-stressed, aged animals, or animals exposed to inflammatory agents, viruses, bacterial pathogens, pathogenic protozoa, radiation or mutagens. Human studies assessed immunomodulatory effects in healthy subjects, or patients with cancers, seasonal allergic rhinitis or aphthous stomatitis. Because of the limited number of human studies, we included some promising open-label controlled trials. Human study durations ranged from four days to seven years; daily doses ranging from 100-5,400 mg were reported to be well-tolerated.

###### 

Immunomodulatory Glucan Extracts: Oral Animal Studies

  ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Source                                Extract                            Animal                                                                      Dose/day                                                               Duration of study   Treatment                                                                                                                                                                                                                    Effects                                                                                                                                                                                                            Reference
  ------------------------------------- ---------------------------------- --------------------------------------------------------------------------- ---------------------------------------------------------------------- ------------------- ---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ ------------
  *Agaricus*\                           α-1,6 and\                         8-week ♀ C3H/He mice (5/group)                                              100 mg/kg IG every 3 days                                              1 month             Healthy animals                                                                                                                                                                                                              ↑ \#s splenic T lymphocytes (Thy1.2, CD4+ and CD8+)                                                                                                                                                                \[[@B24]\]
  (*A. blazei*) *subrufescens*          α-1,4 glucans                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        Aqueous                            7-9-week ♂ Balb/cByJ mice (40/group)                                        1 ml 0.45N, 0.6N, or 3N aqueous extract                                2 months                                                                                                                                                                                                                                         All doses ↑ serum IgG levels, CD3+ T cell populations and PML phagocytic activity                                                                                                                                  \[[@B22]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           7-9-week male Balb/cByJ mice (40/group)                                     1 ml 0.45N, 0.6N, or 3N aqueous extract                                10 weeks            IP injection of OVA at 4 weeks                                                                                                                                                                                               0.6N and 3N ↑ levels of OVA-specific serum IgG 28 days post-immunization; all doses ↑ delayed-type hypersensitivity and TNF-α secreted from splenocytes at 10 weeks; 0.6N ↑ splenocyte proliferation at 10 weeks   

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           5-6 -week ♀ BALB/cHsdOla mice (8/group × 2)                                 One 200 μl extract day 1, orogastric intubation                        1 week              Injected IP fecal solution day 2                                                                                                                                                                                             ↓ CFU in blood of mice with severe peritonitis & improved overall survival rate in all peritonitis groups                                                                                                          \[[@B46]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           6-week BALB/c nu/nu mice (7/group)                                          2.5 mg extract days 20-41, drinking water                              41 days             Injected SC Sp-2 myeloma cells day 1                                                                                                                                                                                         ↓ tumor size & weight after 21 days treatment                                                                                                                                                                      \[[@B65]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        Aqueous, acid treated              6-week ♀ C57BL/6 mice (10/group)                                            20, 100 or 500 μg/ml, drinking water                                   9 days              Injected IP human ovarian cancer cells day 1                                                                                                                                                                                 500 μg/ml ↓ tumor weight                                                                                                                                                                                           \[[@B66]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                       20, 100 or 500 μg/ml, drinking water                                   3 weeks             Injected IV murine lung cancer (3LL) cells                                                                                                                                                                                   100 & 500 μg/ml ↓ \#s metastatic tumors                                                                                                                                                                            

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        Aqueous, with 200 ng/day\          6-week ♀ BALB/c mice (10/group)                                             200 ng days 5-21                                                       3 weeks             Injected Meth A tumor cells day 1                                                                                                                                                                                            ↓ tumor size & weight                                                                                                                                                                                              \[[@B23]\]
                                        β-glucan                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                              2 weeks             Injected Meth A tumor cells                                                                                                                                                                                                  ↑ cytotoxic T lymphocyte activity & spleen cell IFN-α protein                                                                                                                                                      

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                       300 mg                                                                 5 days              Healthy animals                                                                                                                                                                                                              ↑ splenic NK cell activity                                                                                                                                                                                         

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Avena*spp.                           β-glucans (particulate)            6-7 -week ♀ C57BL/6 mice (7/group)                                          3 mg every 48 h, days 1-3                                              1 month             Oral *E. vermiformis*oocytes day 10                                                                                                                                                                                          ↓ *E. vermiformis*fecal oocyte \#s; increased intestinal anti-merozoite IgA; ↓ \# of IL-4-secreting MLN cells                                                                                                      \[[@B42]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                       3 mg on alternating days, days 1-10                                    22 days             Injected IP *Eimeria vermiformis*day 10                                                                                                                                                                                      ↓ *E. vermiformis*fecal oocyte \#s; ↑ anti-merozoite intestinal IgA                                                                                                                                                \[[@B43]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        β-glucans (soluble)                4-week ♂ CD-1 mice (24/group)                                               0.6 mg/ml 68% β-glucan, drinking water                                 1 month             Resting or exercise-stressed (days 8-10) animals administered HSV-1 IN\                                                                                                                                                      ↓ morbidity in resting and exercise-stressed animals; ↓ mortality in exercise-stressed animals; pre-infection, ↑ Mø anti-viral resistance in resting and exercise-stressed animals                                 \[[@B38]\]
                                                                                                                                                                                                                                                  day 10                                                                                                                                                                                                                                                                                                                                                                                                                                          

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                       \~3.5 mg days\                                                                             Resting or exercise-stressed (days 5-10) animals administered HSV-1 IN\                                                                                                                                                      Pre-infection, ↑ Mø antiviral resistance in resting animals                                                                                                                                                        \[[@B41]\]
                                                                                                                                                       1-10, drinking water                                                                       day 10                                                                                                                                                                                                                                                                                                                                                                                                                                          

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           4-week ♂ CD-1 mice (10/group)                                               0.6 mg/ml 68% β-glucan, drinking water                                 10 days             Resting animals or animals exposed to a bout of fatiguing exercise days 8-10 or moderate exercise days 5-10, injected IP with thioglycollate on day 10                                                                       ↑ neutrophil mobilization in resting & moderately exercised animals; ↑ neutrophil respiratory burst activity in resting and fatiguing exercised animals                                                            \[[@B37]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           4-week ♂ CD-1 mice (19-30/group)                                            0.8 mg/ml 50% β-glucan, days\                                          1 month             Resting or exercise-stressed (days 8-10) animals administered IN clodronate-filled liposomes to deplete Mø days 8 & 14 & infected IN with HSV-1 day 10                                                                       ↓ morbidity, mortality, symptom severity in exercise-stressed animals, without Mø depletion                                                                                                                        \[[@B40]\]
                                                                                                                                                       1-10, drinking water                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           4-week ♂ CD-1 mice (20/group)                                                                                                                                          Resting or exercise-stressed (days 8-10) animals administered HSV-1 IN day 10                                                                                                                                                ↓ morbidity in exercise-stressed & resting animals; ↓ mortality in exercise-stressed animals                                                                                                                       \[[@B39]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Ganoderma lucidum*                   Aqueous                            7-week ♂ CD-1 mice (26/group)                                               5% of diet                                                             5 months            Injected IM DMH once a week, weeks 1-10                                                                                                                                                                                      ↓ aberrant crypt foci per colon, tumor size, cell proliferation, nuclear staining of β-catenin                                                                                                                     \[[@B69]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           4-8-week BALB/c mice (10/group)                                             50, 100 or 200 mg/kg, oral                                             10 days             Injected SD Sarcoma 180 cells                                                                                                                                                                                                ↓ of tumor weight was dose dependent: 27.7, 55.8, 66.7%, respectively                                                                                                                                              \[[@B67]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Ganoderma lucidum*(mycelia)          Aqueous                            7-week ♂ F344/Du Crj rats (16/group)                                        1.25% or 2.5% of diet                                                  6 months            Injected SC AOM once a week, weeks2-5                                                                                                                                                                                        Both doses ↓ colonic adenocarcinoma incidence; 2.5% ↓ total tumor incidence; both doses ↓ nuclear staining of β-catenin and cell proliferation                                                                     \[[@B68]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Ganoderma tsugae*                    Aqueous                            8-week ♀ BALB/cByJNarl mice (14/group)                                      0.2-0.4% of diet (young fungi); 0.33 or 0.66% of diet (mature fungi)   5 weeks             Injected IP OVA days 7, 14, 21; aerosolized OVA twice during week 4                                                                                                                                                          In splenocytes, both doses of both extracts ↑ IL-2 and IL-2/IL-4 ratios, 0.2% young extract and 0.66% mature extract ↓ IL-4; in Mø, 0.66% mature extract ↑ IL-1β, both doses of both extracts ↑ IL-6               \[[@B53]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Grifola frondosa*                    D fraction                         Mice: 1) ICR, 2) C3H/HeN, 3) CDF~1~(10/group)                               1.5 mg every other day, beginning day 2                                13 days             Implanted SC: 1) Sarcoma-180, 2) MM-46 carcinoma, or 3) IMC carcinoma cells                                                                                                                                                  ↓ tumor weight & tumor growth rate: 1) 58%, 2) 64%, and 3) 75%, respectively                                                                                                                                       \[[@B71]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           5-week ♂ BALB/c mice (10/group)                                             2 mg,\                                                                 45 days             Injected in the back with 3-MCA, day 1                                                                                                                                                                                       ↓ (62.5%) \# of animals with tumors; ↑ H~2~0~2~production by plasma Mø; ↑ cytotoxic T cell activity                                                                                                                \[[@B72]\]
                                                                                                                                                       days 15-30                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Hordeum vulgare*                     β-1,3;1,4 or β-1,3;1,6-D-glucans   Athymic nu/nu mice\                                                         40 or 400 μg IG for 4 weeks                                            31 weeks            Mice with human xenografts (SKMel28 melanoma, A431 epidermoid carcinoma, BT474 breast carcinoma, Daudi lymphoma, or LAN-1 neuroblastoma) ± mAb (R24, 528, Herceptin, Rituximab, or 3F8, respectively) therapy twice weekly   400 μg + mAb ↓ tumor growth & ↑ survival; higher MW ↓ tumor growth rate for both doses                                                                                                                             \[[@B75]\]
                                                                           (4-12/group)                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        β-1,3;1,4-D-glucans                Athymic BALB/c mice                                                         4, 40, or 400 μg for 3-4 weeks                                         1 month             Mice with neuroblastoma (NMB7, LAN-1, or SK-N-ER) xenografts, ± 3F8 mAb therapy twice weekly                                                                                                                                 40 and 400 μg doses + mAB ↓ tumor growth; 400 μg dose ↑ survival. Serum NK cells required for effects on tumor size                                                                                                \[[@B76]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           C57BL/6 WT and CR3-deficient mice (10/group)                                0.4 mg for 3 weeks                                                     100 days            Injected SC RMA-S-MUC1 lymphoma cells day 1 ± IV 14.G2a or anti-MUC1 mAb every 3rd day                                                                                                                                       ±mAB ↓ tumor diameter; ↑ survival                                                                                                                                                                                  \[[@B73]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        β-glucans                          ♀ Fox Chase ICR immune-deficient (SCID) mice (9/group)                      400 μg days 1-29                                                       50 days             Mice with human (Daudi, EBV-BLCL, Hs445, or RPMI6666) lymphoma xenografts, ± Rituximab mAb therapy twice weekly                                                                                                              +mAB ↓ tumor growth and ↑ survival                                                                                                                                                                                 \[[@B74]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Laminaria digitata*                  Laminarin                          ♂ ICR/HSD mice (3/group)                                                    1 mg                                                                   1 day               Healthy animals                                                                                                                                                                                                              ↑ Mø expression of Dectin-1 in GALT cells; ↑ TLR2 expression in Peyer\'s patch dendritic cells                                                                                                                     \[[@B29]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           ♂ Wistar rats (7/group)                                                     5% of diet days 1-4, 10% of diet days 5-25                             26 days             Injected IP *E. coli*LPS day 25                                                                                                                                                                                              ↓ liver ALT, AST, and LDH enzyme levels; ↑ ED2-positive cells, .↓ peroxidase-positive cells in liver; ↓ serum monocytes, TNF-α, PGE2, NO~2~                                                                        \[[@B44]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Lentinula edodes*                    SME                                6-week nude mice                                                            0.1 ml water with10% SME/10 g body weight days 1-19, 33-50             50 days             Injected SC prostate cancer (PC-3) cells day 1                                                                                                                                                                               ↓ tumor size                                                                                                                                                                                                       \[[@B80]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        β-glucans                          ♀ 3- and 8-week BALB/c mice (15/group)                                      50, 100 or 250 μg                                                      1-2 weeks           Healthy animals                                                                                                                                                                                                              250 μg dose ↑ spleen cell IL-2 secretion                                                                                                                                                                           \[[@B27]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           ♀ 3- and 8-week BALB/c mice (15/group)                                      50, 100 or 250 μg                                                      1-2 weeks           Injected murine mammary carcinoma (Ptas64) cells into mammary fat pads 2 weeks before treatment                                                                                                                              ↓ tumor weight                                                                                                                                                                                                     

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        Lentinan                           6-week ♂ Wistar-Imamichi specific-pathogen free rats (10/group)             1 mg twice weekly                                                      1-2 months          Healthy animals                                                                                                                                                                                                              ↑ T cell \#s, helper-cell \#s & helper/suppressor ratio, ↓ suppressor cell level at 4, but not 8 weeks                                                                                                             \[[@B26]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           5-6-week ♂\                                                                 3 mg, days 1-7                                                         3 weeks             Injected SC K36 murine lymphoma cells day 7                                                                                                                                                                                  ↓ tumor weight; ↑ tumor inhibition rate (94%)                                                                                                                                                                      \[[@B82]\]
                                                                           pre-leukemic AKR mice (10/group)                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           5-6-week athymic mice (10/group)                                                                                                                   5 weeks             Injected SC colon cancer (LoVo and SW48, SW480 and SW620, or SW403 and SW1116) cells day 7                                                                                                                                   ↓ tumor weight, ↑ tumor inhibition rate (\>90%)                                                                                                                                                                    

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           ♂ AKR mice                                                                  3 mg                                                                   1 day               Pre-leukemic mice                                                                                                                                                                                                            ↑ serum IFN-α and TNF-α, peak at 4 h and then back to normal at 24 h; ↑ IL-2 and IL-1α, peak at 2 h and back to normal at 24 h; ↑ CD3+ T, CD4+ T, CD8+ T, B lymphocytes                                            \[[@B81]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Phellinus linteus*                   Aqueous, alcohol-precipitated      6-7-week C57BL/6 mice (10-50/group)                                         200 mg/kg in drinking water                                            1 month             Healthy animals                                                                                                                                                                                                              ↑ production and secretion of IFN-γ by con A stimulated T cells                                                                                                                                                    \[[@B32]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Saccharomyces cerevisiae*            Scleroglucan                       ♂ ICR/HSD mice (3/group)                                                    1 mg one day before challenge (day 1)                                  6 days              IV *Staphylococcus aureus*or *Candida albicans*day 2                                                                                                                                                                         ↑ long-term survival                                                                                                                                                                                               \[[@B29]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        β-1,3;1,6 glucans (particulate)    3 and 8-week ♀ BALB/c mice (15/group)                                       50, 100 or 250 μg                                                      1-2 weeks           Injected murine mammary carcinoma (Ptas64) cells into mammary fat pads 2 weeks before treatment                                                                                                                              ↓ tumor weight                                                                                                                                                                                                     \[[@B27]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                        β-1,3-glucan                                                                                                                                                                                              Healthy animals                                                                                                                                                                                                              All 3 doses ↑ phagocytic activity of blood monocytes & neutrophils & ↑ spleen cell IL-2 secretion                                                                                                                  

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           WT or CCD11b^-/-^C57BL/6 mice (2/group)                                     0.4 mg for 3 weeks                                                     100 days            Injected SC RMA-S-MUC1 lymphoma cells ± 14.G2a or anti-MUC1 mAb IV injection every 3^rd^day                                                                                                                                  ↓ tumor diameter when included with mAb; ↑ survival with and without mAb                                                                                                                                           \[[@B73]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           C57BL/6mice (4/group)                                                       25 mg                                                                  1 week              Healthy animals                                                                                                                                                                                                              ↑ \# intestinal IELs; ↑ \# TCRαβ+, TCR γδ+, CD8+, CD4+, CD8αα+, CD8αβ+ T cells in IELs; ↑ IFN-γ mRNA expression in IELs and spleen                                                                                 \[[@B28]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Sclerotinia sclerotiorum*            SSG                                6-8-week specific pathogen-free ♂ CDF~1~mice (3/group)                      40 or 80 mg/kg days 1-10                                               2 weeks             Healthy animals                                                                                                                                                                                                              10 mg dose ↑ acid phosphatase activity of peritoneal Mø (day 14)                                                                                                                                                   \[[@B30]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                       40, 80 or 160 mg/kg days 2-6                                           35 days             Implanted SC Metha A fibrosarcoma cells day 1                                                                                                                                                                                80 mg dose ↓ tumor weight                                                                                                                                                                                          

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           6-8-week specific pathogen-free ♂ CDF~1~mice (10/group)                     40, 80 or 160 mg/kg days 2-11                                                              Injected ID IMC carcinoma cells day 1                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                           6-8-week specific-pathogen free ♂ mice of BDF1 and C57BL/6 mice (7/group)   0.5, 1, 2, or 4 mg days 1-10                                           2-3 weeks           Injected IV Lewis lung carcinoma (3LL) cells                                                                                                                                                                                 2 mg ↓ \# of 3LL surface lung nodules at 2 weeks                                                                                                                                                                   \[[@B83]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Sclerotium rofsii*                   Glucan phosphate                   ♂ ICR/HSD mice (3/group)                                                    1 mg                                                                   1 day               Healthy animals                                                                                                                                                                                                              ↑ systemic IL-6; ↑ Mø expression of Dectin-1 in GALT cells; ↑ TLR2 expression in dendritic cells from Peyer\'s patches                                                                                             \[[@B29]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Trametes*(*Coriolus*) *versicolor*   PSP                                6-8-week ♂ BALB/c mice (10/group)                                           35 μg days 5-29 in drinking water                                      29 days             Implanted SC Sarcoma-180 cells day 1                                                                                                                                                                                         ↓ tumor growth & vascular density                                                                                                                                                                                  \[[@B94]\]
  ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

###### 

Immunomodulatory Non-Glucan Extracts: Oral Animal Studies

  -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Extract                                                                Source                                                             Animal                                                                                                  Oral dose/day                                                                           Duration   Treatment                                                                                                              Significant effects                                                                                                                                                                                                                       Reference
  ---------------------------------------------------------------------- ------------------------------------------------------------------ ------------------------------------------------------------------------------------------------------- --------------------------------------------------------------------------------------- ---------- ---------------------------------------------------------------------------------------------------------------------- ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- -------------
  Fucoidans                                                              *Cladosiphon okamuranus Tokida*                                    8-week ♀ BALB/c mice, 10/group                                                                          0.05% w/w of diet                                                                       56 days    DSS-induced UC                                                                                                         ↓ disease activity index and myeloperoxidase activity; ↓ \# of B220-positive colonic B cells; ↓ colonic MLN IFN-γ and IL-6 and ↑ IL-10 and TGF-β; ↓ colonic IgG; ↓ colonic epithelial cell IL-6, TNF-α, and TLR4 mRNA expression          \[[@B49]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                         *Undaria pinnatifida*                                              5-week ♀ BALB/c mice (10-12/group)                                                                      5 mg, days 1-14 or 7-14                                                                 2 weeks    Injected HSV into cornea day 7                                                                                         ↓ facial herpetic lesions; ↑ survival, particularly in pre-treated animals                                                                                                                                                                \[[@B45]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                                                                                                                                    10 mg                                                                                   1 week     Administered\                                                                                                          ↑ plasma NK cell activity                                                                                                                                                                                                                 
                                                                                                                                                                                                                                                                                                                                                       5-fluorouracil                                                                                                                                                                                                                                                                                                                                                   

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                                                                                                                                                                                                                                       Injected SC HSV                                                                                                        ↑ cytotoxic splenic T lymphocyte activity                                                                                                                                                                                                 

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                                                                                                                                    0.1 or 0.5 mg                                                                           3 weeks    Injected IP HSV                                                                                                        Both doses ↑ serum neutralizing Ab titers, weeks 2 and 3                                                                                                                                                                                  

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            6-week ♂ ddY mice (5/group)                                                                             50, 100, 200 400 or\                                                                    3 weeks    Injected with Ehrlich carcinoma in back day 14                                                                         200-500 mg/kg ↓ tumor growth                                                                                                                                                                                                              \[[@B116]\]
                                                                                                                                                                                                                                                    500 mg/kg\                                                                                                                                                                                                                                                                                                                                                                                                                                                          
                                                                                                                                                                                                                                                    days 1-28                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            6-week ♂ BALB/c mice (8/group)                                                                          40 mg/kg alternating days\                                                              19 days    Injected IP Meth A fibrosarcoma day 1                                                                                  ↓ tumor growth                                                                                                                                                                                                                            
                                                                                                                                                                                                                                                    7-19                                                                                                                                                                                                                                                                                                                                                                                                                                                                

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Furanose (COLD-FX^®^)                                                  *Panax quinquefolium*                                              Weanling ♂ SD rats (10/group)                                                                           450 or\                                                                                 1 week     Healthy animals                                                                                                        Both doses ↑ spleen Il-2 and IFN-γ production following ConA or LPS stimulation; ↓ proportion of total MLN and Peyer\'s patch CD3+ cells & activated T cells; high dose ↑ spleen cell IL-1β production following 48 h ConA stimulation.   \[[@B33]\]
                                                                                                                                                                                                                                                    900 mg/kg in food                                                                                                                                                                                                                                                                                                                                                                                                                                                   

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Galacto-mannan (partially hydrolyzed guar gum)                         *Cyamopsis tetragonolobus*                                         10-week ♀ BALB/c mice,\                                                                                 5% of diet                                                                              3 weeks    DSS-induced UC at beginning of\                                                                                        ↓ disease activity index scores, ↓ colonic mucosal myeloperoxidase activity & lipid peroxidation; ↓ colonic TNF-α protein levels & mRNA expression up regulated by DSS exposure                                                           \[[@B50]\]
                                                                                                                                            11-15/group                                                                                                                                                                                                week 3                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Galacto-mannans\                                                                                                                          8-month- SD rats, 5/group                                                                               5% of diet                                                                              3 weeks    Older animals                                                                                                          ↓ serum IgG; ↑ MLN lymphocyte IgA, IgM and IgG production                                                                                                                                                                                 \[[@B36]\]
  (guar gum)                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Glucomannan (KS-2)                                                     *Lentinula edodes*                                                 DD1 mice (10-20/group)                                                                                  140 mg/kg days\                                                                         50 days    Injected IP Ehrlich ascites tumor cells day 1                                                                          ↑ survival                                                                                                                                                                                                                                \[[@B84]\]
                                                                                                                                                                                                                                                    2-13                                                                                                                                                                                                                                                                                                                                                                                                                                                                

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                                                                                                                                    0.1, 1, 10, or 100 mg/kg dose days 2-13                                                 100 days   Injected Sarcoma-180 tumor cells\                                                                                      1, 10, and 100 mg/kg doses ↑ survival                                                                                                                                                                                                     
                                                                                                                                                                                                                                                                                                                                                       day 1                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Heteroglycan (ATOM)                                                    *A. subrufescens*                                                  Mice (10/group): 1) 5-week ♂ Swiss/NIH; 6 week- ♀ DS mice; 3) 8-week ♀ BALB/c nude; 4) 5-week C3H/HcN   100 or\                                                                                 8 weeks    Implanted SC 1) Sarcoma-180, 2) Shionogi carcinoma 42, 3) Meth A fibrosarcoma, or 4) Ehrlich ascites carcinoma cells   Both doses ↓ Sarcoma-180 tumor size at 4 weeks & ↑ survival; 300 mg/kg ↑ peritoneal macrophage and C3-positive cells; 300 mg/kg ↓ Shionogi and Meth A tumor sizes at 4 weeks. Both doses ↑ survival of Ehrlich ascites mice               \[[@B93]\]
                                                                                                                                                                                                                                                    300 mg/kg\                                                                                                                                                                                                                                                                                                                                                                                                                                                          
                                                                                                                                                                                                                                                    days 2-11                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Heteroglycan (LBP~3p~)                                                 *Lycium barbarum*                                                  ♂ Kunming mice (10/group)                                                                               5, 10 or\                                                                               10 days    Injected SC Sarcoma-180 cells                                                                                          5 & 10 mg/kg ↑ thymus index; all doses ↓ weight, ↓ lipid peroxidation in serum, liver and spleen & ↑ spleen lymphocyte proliferation, cytotoxic T cell activity, IL-2 mRNA                                                                \[[@B91]\]
                                                                                                                                                                                                                                                    20 mg/kg                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Heteroglycan (PNPS-1)                                                  *Pholiota nameko*                                                  SD rats (5/group)                                                                                       100, 200 or 400 mg/kg days 1-8                                                          8 days     Implanted SC cotton pellets in scapular region\                                                                        ↓ granuloma growth positively correlated with dose: 11%, 18% and 44%, respectively                                                                                                                                                        \[[@B55]\]
                                                                                                                                                                                                                                                                                                                                                       day 1                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Heteroglycan (PG101)                                                   *Lentinus lepideus*                                                8-10-week ♀ BALB/c mice (3/group)                                                                       10 mg                                                                                   24 days    6 Gy gamma irradiation                                                                                                 ↑ colony forming cells, granulocyte CFUs/Mø, erythroid burst-forming units, and myeloid progenitor cells in bone marrow; induced proliferation of granulocyte progenitor cells in bone marrow; ↑ serum levels of GM-CSF, IL-6, IL-1β      \[[@B92]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Mixed poly-saccharides (Ambrotose^®^or Advanced Ambrotose^®^powders)   *Aloe barbadensis*, *Larix*spp, and other plant poly-saccharides   ♂ SD rats (10/group)                                                                                    37.7 or 377 mg/kg Ambrotose^®^powder or 57.4 or 574 mg/kg Advanced Ambrotose^®^powder   2 weeks    5% DSS in drinking water beginning day 6                                                                               574 mg/kg Advanced Ambrotose powder ↓ DAI scores; 377 mg/kg Ambrotose complex & both doses Advanced Ambrotose powder ↑ colon length and ↓ blood monocyte count                                                                            \[[@B52]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Pectin                                                                 *Pyrus pyrifolia*                                                  6-8-week ♂ BALB/c mice (11/group)                                                                       100 μg\                                                                                 22 days    Injected IP OVA day 7, provoked with OVA aerosol day 21                                                                bronchial fluid:↓ IFN-γ & ↑ IL-5; splenic cells: ↑ IFN-γ, ↓ IL-5; normalized pulmonary histopathological changes; ↓ serum IgE                                                                                                             \[[@B54]\]
                                                                                                                                                                                                                                                    days 1-7                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Pectins (bupleurum 2IIc)                                               *Bupleurum falcatum*                                               6-8-week ♀ specific-pathogen-free C3H/HeJ mice                                                          250 mg/kg                                                                               1 week     Healthy animals                                                                                                        ↑ spleen cell proliferation                                                                                                                                                                                                               \[[@B35]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Pectins (highly methoxylated)                                          *Malus*spp.                                                        8-month- SD rats (5/group)                                                                              5% of diet vs. cellulose control                                                        3 weeks    Older animals                                                                                                          ↑ MLN lymphocyte IgA & IgG                                                                                                                                                                                                                \[[@B36]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Pectins                                                                Citrus spp.                                                        5-week ♀ F344 rats (30/group)                                                                           15% of diet                                                                             34 weeks   Injected SC AOM once a week, weeks 4-14                                                                                ↓ colon tumor incidence                                                                                                                                                                                                                   \[[@B86]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                         *Malus*spp.                                                        5-week ♀ BALB/c mice (6/group)                                                                          5% of diet                                                                              2 weeks    Healthy animals                                                                                                        ↑ fecal IgA and MLN CD4+/CD8+ T lymphocyte ratio & IL-2 & IFN-γ secretion by ConA-stimulated MLN lymphocytes                                                                                                                              \[[@B51]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            5-week ♀ BALB/c mice (6/group)                                                                          5% of diet days 5-19 vs. cellulose control                                              19 days    DSS-induced UC days 1-5                                                                                                Significantly increased MLN lymphocytes IgA, and significantly decreased IgE; significantly decreased ConA-stimulated IL-4 and IL-10                                                                                                      

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            4-week ♂ Donryu rats (20-21/group)                                                                      20% of diet                                                                             32 weeks   Injected SC AOM once a week,\                                                                                          ↓ colon tumor incidence                                                                                                                                                                                                                   \[[@B85]\]
                                                                                                                                                                                                                                                                                                                                                       weeks 2-12                                                                                                                                                                                                                                                                                                                                                       

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            4-week ♂ Donryu rats (19-20/group)                                                                      10 or 20% of diet                                                                       32 weeks   Injected SC AOM once a week,\                                                                                          Both doses ↓ colon tumor incidence; 20% ↓ tumor occupied area & ↓ portal blood and distal colon PGE~2~                                                                                                                                    \[[@B90]\]
                                                                                                                                                                                                                                                                                                                                                       weeks 2-12                                                                                                                                                                                                                                                                                                                                                       

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Pectins (modified)                                                     Citrus spp.                                                        2-4-month BALB/c mice (9-10/group)                                                                      0.8 or 1.6 mg/ml drinking water,\                                                       20 days    Injected SC with 2 × 2 mm section of human colon-25 tumor on day 1                                                     Both doses ↓ tumor size                                                                                                                                                                                                                   \[[@B87]\]
                                                                                                                                                                                                                                                    days 8-20                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            NCR nu/nu mice (10/group)                                                                               1% (w/v) drinking water                                                                 16 weeks   Orthotopically injected human breast carcinoma cells (MDA-MB-435) into mammary fat pad on day 7                        ↓ tumor growth rate & volume at 7 weeks, lung metastases at 15 weeks, \# of blood vessels/tumor at 33 days post-injection                                                                                                                 \[[@B89]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            NCR nu/nu mice (10/group)                                                                               1% (w/v) drinking water                                                                 7 weeks    Injected human colon carcinoma cells (LSLiL6) into cecum on day 7                                                      ↓ tumor weights and metastases to the lymph nodes and liver                                                                                                                                                                               

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                            SD rats (7-8/group)                                                                                     0.01%, 0.1% or 1.0% wt/vol of drinking water, days 4-30                                 1 month    Injected SC MAT-LyLu rat prostate cancer cells                                                                         0.1% and 1.0% ↓ lung metastases; 1.0% ↓ lymph node disease incidence                                                                                                                                                                      \[[@B88]\]
  -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

###### 

Immunomodulatory Polysaccharide-Rich Plant Powders: Oral Animal Studies

  -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Source                                                 Animal                                                            Oral dose/day                                             Duration    Treatment                                                                                                                      Significant effects                                                                                                                                        Reference
  ------------------------------------------------------ ----------------------------------------------------------------- --------------------------------------------------------- ----------- ------------------------------------------------------------------------------------------------------------------------------ ---------------------------------------------------------------------------------------------------------------------------------------------------------- ------------
  *Agaricus*(*A. blazei*) *subrufescens*(fruit bodies)   6-week ♂ C57BL/6, C3H/HeJ and BALB/c mice (3/group)               16, 32 or 64 mg                                           2 weeks     Healthy animals                                                                                                                32 and 64 mg ↑ liver mononuclear cell cytotoxicity                                                                                                         \[[@B25]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

  *Grifola frondosa*                                     6-week ♀ ICR mice (10-15/group)                                   5% of diet                                                36 weeks    Oral N-butyl-N\'-butanolnitrosamine daily for first 8 weeks                                                                    ↓ \#s of animals with bladder tumors; ↓ tumor weight; ↑ peritoneal Mø chemotactic activity, splenic lymphocyte blastogenic response & cytotoxic activity   \[[@B70]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

  *Laminaria angustata*                                  Weanling SD rats (58/group)                                       5% of diet                                                26 weeks    IG DMBA, beginning of week 5                                                                                                   ↑ time to tumor development and ↓ \# of adenocarcinomas in adenocarcinoma-bearing animals                                                                  \[[@B77]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

  *Lentinula*(*Lentinus*) *edodes*                       6-week ♀ ICR mice (10-17/group)                                   5% of diet                                                36 weeks    Oral BBN daily for first 8 weeks                                                                                               ↓ \# of animals with bladder tumors; ↓ tumor weight; ↑ Mø chemotactic activity, splenic lymphocyte blastogenic response, cytotoxic activity                \[[@B70]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                         7-8 -week ♂ Swiss mice (10/group)                                 1%, 5% or 10% of diet of 4 different lineages days 1-15   16 days     Injected IP N-ethyl-N-nitrosourea day 15                                                                                       All 3 doses of one lineage and the 5% dose of two other lineages ↓ \#s of micronucleated bone marrow polychromatic erythrocytes                            \[[@B79]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

  *Lentinula edodes*(fruit bodies)                       5-week ♀ ICR mice\                                                10%, 20% or 30% of diet                                   25 days     Injected IP Sarcoma-180 ascites                                                                                                All 3 doses ↓ Sarcoma-180 tumor weight                                                                                                                     \[[@B78]\]
                                                         (14/group × 2)                                                                                                                                                                                                                                                                                                                                                                                                                    

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                         Mice: 1) CDF~1~; 2) C3H; 3) BALB/c; 4,5) C57BL/6N (9/group × 3)   20% of diet                                               25 days     Injected SC 1) IMC carcinoma, 2) MM-46 carcinoma, 3) Meth-A fibrosarcoma, 4) B-16 melanoma, or 5) Lewis lung carcinoma cells   ↓ growth of MM-46, B-16, Lewis lung, and IMC tumors; ↑ lifespan in Lewis lung and MM-46 animals                                                            

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                         ICR mice (14/group × 2)                                           20% of diet days 1-7, days 7-31 or days 14-31             31 days     Injected IP Sarcoma-180 ascites                                                                                                ↓ tumor weight & growth when fed days 7-31 or 14-31                                                                                                        

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                         Mice: 1) CDF~1~; 2) C3 H (5/group × 4)                            20% of diet                                               7-12 days   Injected SC: 1) IMC carcinoma or 2) MM-46 carcinoma cells                                                                      ↑ spreading rate of activated Mø ↑ phagocytic activity                                                                                                     

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

  *Phellinus linteus*                                    4-week ♂ ICR mice (10/group)                                      2 mg                                                      1 month     Healthy animals                                                                                                                ↓ serum & splenocyte IgE production; ↑ proportion of splenic CD4^+^T cells & splenocyte IFN-γ production                                                   \[[@B31]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           

  *Pleurotus ostreatus*                                  6-week ♀ ICR mice\                                                5% of diet                                                36 weeks    Oral BBN daily for first 8 weeks                                                                                               ↓ \#s of animals with bladder tumors; ↓ tumor weight; ↑ plasma Mø chemotactic activity, splenic lymphocyte blastogenic response, cytotoxic activity        \[[@B70]\]
                                                         (10-20/group)                                                                                                                                                                                                                                                                                                                                                                                                                     
  -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

###### 

Immunomodulatory Polysaccharide Products: Oral Human Studies

  ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Extract                          Source                             Study design                                   Population                                                                         N (experimental/control)               Dose/day                                                               Dura-tion                 Significant effects                                                                                                                                                                                                                                         Reference
  -------------------------------- ---------------------------------- ---------------------------------------------- ---------------------------------------------------------------------------------- -------------------------------------- ---------------------------------------------------------------------- ------------------------- ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- ------------
  Arabino-galactans                *Larix occidentalis*               Randomized, double-blind, placebo-controlled   Healthy adults                                                                     8/15                                   4 g                                                                    6 weeks                   ↑ % CD8+ lymphocytes & blood lymphocyte proliferation                                                                                                                                                                                                       \[[@B18]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Arabino-galactans (ResistAid™)                                                                                     Healthy adults given pneumococcal vaccinations day 30                              21/24                                  4.5 g                                                                  72 days                   ↑ plasma IgG subtypes                                                                                                                                                                                                                                       \[[@B19]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Fucoidans                        *Undaria pinnatifida*sporophylls   Randomized, single-blind, placebo-controlled   Healthy adults                                                                     25 (75% fucoidan, 6 (10% fucoidan)/6   3 g                                                                    12 days                   75% fucoidan: ↓ \#s blood leukocytes, lymphocytes\' ↑ plasma stromal derived factor-1, IFN-γ, CD34+ cells; ↑ % CXCR4-expressing CD34+ cells                                                                                                                 \[[@B21]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Furanose extract (Cold-FX^®^)    *Panax quinque-folium*             Randomized, double-blind, placebo-controlled   Healthy older adults given influenza immunization at the end of week 4             22/21                                  400 mg                                                                 4 months                  During weeks 9-16, ↓ incidence of acute respiratory illness, symptom duration                                                                                                                                                                               \[[@B20]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Glucans                          *Agaricus subru-fescens*           Randomized, double-blind, placebo-controlled   Cervical, ovarian or endometrial cancer patients receiving 3 chemotherapy cycles   39/61                                  5.4 g (estimated)                                                      6 weeks                   ↑ NK cell activity, ↓ chemotherapy side effects                                                                                                                                                                                                             \[[@B64]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Glucans\                         Not identified                     Placebo-controlled                             Recurrent aphthous stomatitis patients                                             31/42                                  20 mg                                                                  20 days                   ↑ PBL lymphocyte proliferation,↓ Ulcer Severity Scores                                                                                                                                                                                                      \[[@B48]\]
  (β-1,3;1,6)                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Glucans\                         *S. cerevisiae*                    Randomized, double-blind, placebo-controlled   Adults with seasonal allergic rhinitis                                             12/12                                  20 mg                                                                  12 weeks                  30 minutes after nasal allergen provocation test, nasal lavage fluid: ↓ IL-4, IL-5, % eosinophils, ↑ IL-12                                                                                                                                                  \[[@B47]\]
  (β-1,3;1-6)                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Glucans (PSK)                    *Trametes versicolor*              Randomized, controlled                         Patients with curatively resected colorectal cancer receiving chemotherapy         221/227                                200 mg                                                                 3-5 years                 ↑ disease-free survival and overall survival                                                                                                                                                                                                                \[[@B56]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                      Controlled                                     Post-surgical colon cancer patients receiving chemotherapy                         123/121                                3 g for 4 weeks, alternating with 10 4-week courses of chemo-therapy   7 years                   ↑ survival from cancer deaths; no difference in disease-free or overall survival                                                                                                                                                                            \[[@B57]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                     Post-surgical colorectal cancer patients receiving chemotherapy                    137/68                                 3 g daily                                                              2 years                   ↑survival in stage III patients; ↓ recurrence in stage II & III patients                                                                                                                                                                                    \[[@B58]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                     Post-surgical gastric cancer patients receiving chemotherapy                       124/129                                3 g for 4 weeks, alternating with 10 4-week courses of chemo-therapy   5-7 years                 ↑ 5-year disease-free survival rate, overall 5-year survival                                                                                                                                                                                                \[[@B59]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                                                                     Pre-surgical gastric or colorectal cancer patients                                 16 daily; 17 every other day/13        3 g daily or on alternate days before surgery                          \<14 days or 14-36 days   ≥14 day treatment: ↑ peripheral blood NK cell activity, PBL cytotoxicity, proportion of PBL helper cells; ↓ proportion of PBL inducer cells; \<14 day treatment: ↑ PBL response to PSK and Con A, proportion of regional node lymphocyte suppressor cells   \[[@B62]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                      Randomized, double-blind, placebo-controlled   Post-surgical stage III-IV colorectal cancer patients                              56/55                                  3 g for 2 months, 2 g for 22 months, 1 g thereafter                    8-10 years                ↑ remission & survival rates                                                                                                                                                                                                                                \[[@B61]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

                                                                      Controlled                                     Post-surgical stage III gastric cancer patients receiving chemotherapy             32/21                                  3 g                                                                    1 year                    ↑ survival time                                                                                                                                                                                                                                             \[[@B60]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            

  Glucans (PSP)                    *Trametes versicolor*              Randomized, double-blind, placebo-controlled   Conventionally-treated stage III-IV non-small cell lung cancer patients            34/34                                  3.06 g                                                                 1 month                   ↑ blood IgG & IgM, total leukocyte and neutrophil counts, % body fat; ↓ patient withdrawal due to disease progression                                                                                                                                       \[[@B63]\]
  ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

###### 

Immunomodulatory Polysaccharide Products: Composition and Structure

  --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Source                                                                                                                                                                          Category                                  Features                                                                                                                                                                                                     MW                                                                       Monosaccharide composition                                                                                                      Reference
  ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- ----------------------------------------- ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ ------------------------------------------------------------------------ ------------------------------------------------------------------------------------------------------------------------------- --------------------------------------------
  *Agaricus subrufescens*(*A. blazei*)                                                                                                                                            Extract                                   β-1,6-D-glucan                                                                                                                                                                                               10,000                                                                   NA                                                                                                                              \[[@B66]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Agaricus subrufescens*(fruit body)                                                                                                                                             Extract                                   α-1,6- and α-1,4 glucans with β-1,6-glucopyranosyl backbone (629.2 mcg/mg polysaccharides, 43.5 mcg/mg protein)                                                                                              170,000                                                                  glucose                                                                                                                         \[[@B24]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                            α-1,4 glucans & β-1,6 glucans with β-1,3 side branches; α-1,6 glucans; β-1,6; 1-3 glucans, β-1,4 glucans; β-1,3 glucans; β-1,6; α-1,3 glucans; riboglucans, galactoglucomannans, β-1,2; β-1,3 glucomannans   NA                                                                       glucose, mannose, galactose, ribose                                                                                             \[[@B25],[@B117],[@B118]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Agaricus subrufescens*(mycelia)                                                                                                                                                Extract (ATOM)                            β-1,6-D-glucan, protein complex, 5% protein                                                                                                                                                                  100,000-1,000,000                                                        glucose, mannose, galactose, ribose                                                                                             \[[@B93]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Aloe barbadensis*(leaf gel)                                                                                                                                                    Whole tissue                              Dry weight: 10% polysaccharides; acemannan, aloemannan, aloeride, pectic acid, galactans, arabinans, glucomannans                                                                                            average 2,000,000                                                        mannose, glucose, galactose, arabinose, xylose, rhamnose                                                                        \[[@B119],[@B120]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract (aloemannan)                      neutral partially acetylated glucomannan, mainly β-1,4-mannans                                                                                                                                               \>200,000                                                                mannose, glucose                                                                                                                \[[@B121]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract (aloeride)                        NA                                                                                                                                                                                                           4,000,000-7,000,000                                                      37% glucose, 23.9% galactose, 19.5% mannose, 10.3% arabinose                                                                    \[[@B122]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract (acemannan)                       β-1,4 acetylated mannan                                                                                                                                                                                      80,000                                                                   mannose                                                                                                                         \[[@B123]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Aloe barbadensis*, (leaf gel), *Larix*sp. (bark), *Anogeissus latifolia*(bark), *Astragalus gummifer*(stem), *Oryza sativa*(seed), *glucosamine*                               Extracts (Ambrotose^®^powder)             β-1,4 acetylated mannan, arabinogalactans, polysaccharide gums, rice starch, 5.4% protein                                                                                                                    57.3% ≥ 950,000; 26.4% \< 950,000 and ≥80,000; 16.3% ≤ 10,000            mannose, galactose, arabinose, glucose, galacturonic acid, rhamnose, xylose, fructose, fucose, glucosamine, galacturonic acid   (unpublished data, Mannatech Incorporated)

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Aloe barbadensis*(leaf gel), *Larix*sp. (bark), *Undaria pinnatifida*(frond), *Anogeissus latifolia*(bark), *Astragalus gummifer*(stem), *Oryza sativa*(seed), *glucosamine*   Extracts (Advanced Ambrotose^®^powder)    β-1,4 acetylated mannan, arabinogalactans, polysaccharide gums, fucoidans, rice starch, 6% protein, 1% fatty acids                                                                                           13% = 1,686,667; 46% = 960,000 30% \<950,000 and ≥70,000; 11% ≤ 10,000                                                                                                                                   

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Avena*spp. (seed endosperm)                                                                                                                                                    Extract                                   β-1,3;1,4 particulate (1-3 μ) glucans                                                                                                                                                                        1,100,000                                                                glucose                                                                                                                         \[[@B43]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Avena*spp. (seed)                                                                                                                                                              Extract                                   β-1,4,1,3 particulate glucans (linear chains of β-D-glycopyranosyl units; 70% β 1-4 linked)                                                                                                                  2,000,000                                                                NA                                                                                                                              \[[@B41],[@B124]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Buplerum falcatum*(root)                                                                                                                                                       Extract (bupleuran 2IIc)                  6 linked galactosyl chains with terminal glucuronic acid substituted to β-galactosyl chains                                                                                                                  NA                                                                       galactose, glucuronic acid, rhamnose                                                                                            \[[@B35]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  Citrus spp. (fruit)                                                                                                                                                             Extract                                   α-1,4-linked partially esterified D-anhydrogalacturonic acid units interrupted periodically with 1,2-rhamnose                                                                                                70,000-100,000                                                           galactose, galacturonic acid, arabinose, glucose, xylose, rhamnose                                                              \[[@B125]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Cladosiphon okamuranus*(frond)                                                                                                                                                 Extract                                   α-1,3-fucopyranose sulfate                                                                                                                                                                                   56,000                                                                   fucose:glucuronic acid (6.1:1.0)                                                                                                \[[@B126]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Cordyceps sinensis*(mycelia)                                                                                                                                                   Extract                                   β-1,3-D-glucan with 1,6-branched chains                                                                                                                                                                      NA                                                                       NA                                                                                                                              \[[@B127]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Cyamopsis tetragonolobus*(seed)                                                                                                                                                Extract (guar gum)                        Main chain of β-1,4-mannopyranosyl units with α-galactopyranosyl units                                                                                                                                       220,000                                                                  mannose, galactose                                                                                                              \[[@B36],[@B128]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract (partially-hydrolyzed guar gum)   NA                                                                                                                                                                                                           20,000                                                                   mannose, galactose                                                                                                              \[[@B50]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Flammulina velutipes*                                                                                                                                                          Extract                                   NA                                                                                                                                                                                                           NA                                                                       glucose, mannose, galactose                                                                                                     \[[@B117]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Flammulina velutipes*(fruit body)                                                                                                                                              Extract                                   β-1,3 glucan                                                                                                                                                                                                 NA                                                                       glucose                                                                                                                         \[[@B129]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Ganoderma lucidum*                                                                                                                                                             Whole tissue                              Linear β-1,3-glucans with varying degrees of\                                                                                                                                                                400,000-1,000,000                                                        glucose, galactose, mannose, xylose, uronic acid                                                                                \[[@B130]\]
                                                                                                                                                                                                                            D-glucopyranosyl branching, β-glucan/protein complexes, heteropolysaccharides                                                                                                                                                                                                                                                                                                                                         

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract                                   NA                                                                                                                                                                                                           7,000-9,000                                                              NA                                                                                                                              \[[@B67]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Ganoderma lucidum*(fruit body)                                                                                                                                                 Extract                                   NA                                                                                                                                                                                                           7,000-9,000                                                              NA                                                                                                                              

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                            β-linked heteroglycan peptide                                                                                                                                                                                513,000                                                                  fructose, galactose, glucose, rhamnose, xylose (3.167:\                                                                         \[[@B15]\]
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  0.556:6.89:0.549:3.61)                                                                                                          

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Ganoderma tsugae*                                                                                                                                                              Extract                                   55.6% carbohydrates (12.5% polysaccharides); 12% triterpenes, 1.7% sodium, 0.28% protein, 0% lipid                                                                                                           NA                                                                       NA                                                                                                                              \[[@B53]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Ginkgo biloba*(seed)                                                                                                                                                           Extract                                   89.7% polysaccharides                                                                                                                                                                                        NA                                                                       glucose, fructose, galactose, rhamnose                                                                                          \[[@B131]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Grifola frondosa*                                                                                                                                                              Whole tissue                              β-1,3; 1, 6-glucans, α-glucans, mannoxyloglucans, xyloglucans, mannogalactofucans                                                                                                                            NA                                                                       glucose, fucose, xylose, mannose, galactose                                                                                     \[[@B117]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Grifola frondosa*(fruit body)                                                                                                                                                  Extract\                                  β-1,6-glucan with β-1,3 branches, 30% protein                                                                                                                                                                NA                                                                       glucose                                                                                                                         \[[@B132]\]
                                                                                                                                                                                  (D fraction)                                                                                                                                                                                                                                                                                                                                                                                                                                                    

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract\                                  β-1,6-D-glucan with α-1,4 branches, 35% protein                                                                                                                                                              550,000-558,000                                                          glucose                                                                                                                         
                                                                                                                                                                                  (X fraction)                                                                                                                                                                                                                                                                                                                                                                                                                                                    

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Hordeum*spp. (seed)                                                                                                                                                            Extract                                   β-1,3;1,4-and β-1,3;1,6-D-glucans                                                                                                                                                                            45,000-404,000                                                           glucose                                                                                                                         \[[@B75]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                            Primarily linear β-1,3;1,4- glucans                                                                                                                                                                          NA                                                                       glucose                                                                                                                         \[[@B124]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Laminaria*spp.\                                                                                                                                                                Extract (laminarin)                       β-1,3;1-6 glucan                                                                                                                                                                                             7,700                                                                    glucose                                                                                                                         \[[@B29]\]
  (frond)                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                            β-1,3 glucan with some β-1,6 branches and a small amount of protein                                                                                                                                          4,500-5,500                                                              glucose                                                                                                                         \[[@B44]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract                                   Fucoidan                                                                                                                                                                                                     NA                                                                       NA                                                                                                                              \[[@B133]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Larix occidentalis*(bark)                                                                                                                                                      Extract                                   β-1,3;1,6-D-galactans with arabinofuranosyl and arabinopyranosyl side chains                                                                                                                                 19,000-40,000                                                            galactose:arabinose (6:1), uronic acid                                                                                          \[[@B128],[@B134]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Lentinula edodes*                                                                                                                                                              Extract (SME)                             β-1,3-glucans (4-5%), α-1,4-glucan (8-10%), protein (11-14%)                                                                                                                                                 NA                                                                       glucose                                                                                                                         \[[@B80]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract                                   β-glucan                                                                                                                                                                                                     1,000                                                                    glucose                                                                                                                         \[[@B27]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Whole tissue                              Linear β-1,3-glucans, β-1,4;1,6-glucans, heterogalactan                                                                                                                                                      NA                                                                       glucose, galactose, mannose, fucose, xylose                                                                                     \[[@B135]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract (lentinan)                        β-1,3-glucan with 2 β-1,6 glucopyranoside branchings for every 5 β-1,3-glucopyranoside linear linkages                                                                                                       500,000                                                                  glucose                                                                                                                         \[[@B136]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Lentinula edodes*(fruit body)\                                                                                                                                                 Extract (lentinan)                        Neutral β-1,3-D glucan with two β-1,6 glucoside branches for every five β-1,3 units                                                                                                                          400,000-800,000                                                          glucose                                                                                                                         \[[@B137]\]
  *Lentinula edodes*                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract\                                  Peptide units and mannan connected by α-glycosidic bonds                                                                                                                                                     60,000-90,000                                                            mannose, glucose                                                                                                                
                                                                                                                                                                                  (KS-2)                                                                                                                                                                                                                                                                                                                                                                                                                                                          

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Lentinula edodes*(mycelia or fruit body)                                                                                                                                       Extract                                   Triple helical β-1,3-D glucan with β-1,6 glucoside branches                                                                                                                                                  1,000,000                                                                glucose                                                                                                                         \[[@B3]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Lentinula edodes*(mycelia)                                                                                                                                                     Extract\                                  44% sugars, 24.6% protein                                                                                                                                                                                    \~1,000,000                                                              xylose, arabinose, glucose, galactose, mannose, fructose                                                                        \[[@B3]\]
                                                                                                                                                                                  (LEM)                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract (PG101)                           72.4% polysaccharides, 26.2% protein, 1.4% hexosamine                                                                                                                                                        NA                                                                       55.6% glucose, 25.9% galactose, 18.5% mannose                                                                                   \[[@B138]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Lycium barbarum*                                                                                                                                                               Whole tissue                              α-1,4;1,6-D-glucans, lentinan, β-1,3;1,6 heteroglucans, heterogalactans, heteromannans, xyloglucans                                                                                                          NA                                                                       glucose, galactose, mannose, xylose                                                                                             \[[@B139]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Lycium barbarum*(fruit body)                                                                                                                                                   Extract\                                  88.36% sugars, 7.63% protein                                                                                                                                                                                 157,000                                                                  galactose, glucose, rhamnose, arabinose, mannose, xylose (molar ratio of 1:2.12:1.25:1.10:1.95:1.76)                            \[[@B91]\]
                                                                                                                                                                                  (LBP~3p~)                                                                                                                                                                                                                                                                                                                                                                                                                                                       

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Panax quinquefolium*(root)                                                                                                                                                     Extract                                   Poly-furanosyl-pyranosyl saccharides                                                                                                                                                                         NA                                                                       arabinose, galactose, rhamnose, galacturonic acid, glucuronic acid                                                              \[[@B33]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                            NA                                                                                                                                                                                                           NA                                                                       glucose, mannose, xylose                                                                                                        \[[@B140]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract\                                  90% poly-furanosyl-pyranosyl-saccharides                                                                                                                                                                     NA                                                                       furanose                                                                                                                        \[[@B20]\]
                                                                                                                                                                                  (Cold-fX^®^)                                                                                                                                                                                                                                                                                                                                                                                                                                                    

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Phellinus linteus*(fruit body)                                                                                                                                                 Extract                                   α- and β-linked 1,3 acidic proteoglycan with 1,6 branches                                                                                                                                                    150,000                                                                  glucose, mannose, arabinose, xylose                                                                                             \[[@B141]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Phellinus linteus*(mycelia)                                                                                                                                                    Extract                                   83.2% polysaccharide (4.4% β-glucan), 6.4% protein, 0.1% fat                                                                                                                                                 NA                                                                       glucose                                                                                                                         \[[@B142]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Pholiota nameko*(fruit body)                                                                                                                                                   Extract (PNPS-1)                          NA                                                                                                                                                                                                           114,000                                                                  mannose, glucose, galactose, arabinose, xylose (molar ratio of 1:8.4:13.6:29.6:6.2)                                             \[[@B55]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Pleurotus ostreatus*(mycelia)                                                                                                                                                  Extract                                   β-1,3;1,6-D-glucans                                                                                                                                                                                          316,260                                                                  glucose                                                                                                                         \[[@B143]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Saccharomyces cerevisiae*                                                                                                                                                      Extract (WGP)                             Particulate β-1,3;1,6-D-glucan                                                                                                                                                                               NA                                                                       glucose                                                                                                                         \[[@B144]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract                                   β-glucans with β-1,6 branches with a β-1,3 regions                                                                                                                                                           NA                                                                       glucose                                                                                                                         \[[@B124]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract\                                  soluble β-1,3-D-glucan with β-1,3 side chains attached with β-1,6 linkages                                                                                                                                   20,000                                                                   glucose                                                                                                                         \[[@B145]\]
                                                                                                                                                                                  (SBG)                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Sclerotinia sclerotiorum*(mycelia)                                                                                                                                             Extract\                                  β-1,3-D-glucan, \<1% protein (\>98% polysaccharide)                                                                                                                                                          NA                                                                       glucose                                                                                                                         \[[@B83]\]
                                                                                                                                                                                  (SSG)                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Sclerotium rofsii*                                                                                                                                                             Extract (scleroglucan)                    β-1,3;1,6 glucan                                                                                                                                                                                             1,000,000                                                                glucose                                                                                                                         \[[@B29]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Trametes versicolor*(fruit body)                                                                                                                                               Extract\                                  α-1,4, β-1,3 glucans, 10% peptides                                                                                                                                                                           100,000                                                                  glucose, arabinose, mannose, rhamnose                                                                                           \[[@B146]\]
                                                                                                                                                                                  (PSP)                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Trametes versicolor*(mycelia)                                                                                                                                                  Extract\                                  β-1,4;1,3;1,6-D-glucans, protein                                                                                                                                                                             94,000                                                                   glucose (74.6%), mannose (15.5%), xylose (4.8%), galactose (2.7%), fucose (2.4%)                                                \[[@B137],[@B147]\]
                                                                                                                                                                                  (PSK)                                                                                                                                                                                                                                                                                                                                                                                                                                                           

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  *Undaria pinnatifida*(sporophyll)                                                                                                                                               Extract                                   Galactofucan sulfate                                                                                                                                                                                         9,000                                                                    fucose:galactose 1.0:1.1                                                                                                        \[[@B148]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                            Galactofucan sulfate                                                                                                                                                                                         63,000                                                                   fucose:galactose:gluc-uronic acid (1.0:1.0:0.04)                                                                                \[[@B149]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                                                            β-1,3-galactofucan sulphate                                                                                                                                                                                  38,000                                                                   fucose, galactose                                                                                                               \[[@B150]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

  Unidentified source                                                                                                                                                             Extract (modified citrus pectin)          NA                                                                                                                                                                                                           10,000                                                                   galactose, rhamnose, uronic acid                                                                                                \[[@B125]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  

                                                                                                                                                                                  Extract (highly methoxylated pectin)      NA                                                                                                                                                                                                           200,000                                                                  NA                                                                                                                              \[[@B36]\]
  --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

###### 

Safety of Immunomodulatory Polysaccharide Products Following Oral Intake

  ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Category            Source                                                      Test group                                           Test                              Design                                                                                Results                                                                                                                                                        Equivalent human dose\*   Reference
  ------------------- ----------------------------------------------------------- ---------------------------------------------------- --------------------------------- ------------------------------------------------------------------------------------- -------------------------------------------------------------------------------------------------------------------------------------------------------------- ------------------------- ------------------------------------
  Arabino-galactans   *Argemone mexicana*(arabinogalactan protein)                Pregnant rats                                        Develop-mental toxicity           250, 500, or 1,00 mg/kg, gestational days 5-19                                        No developmental toxicity: NOAEL = 1 g/kg                                                                                                                      68 g                      \[[@B151]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  ♀ and ♂ rats                                         Fertility                         250, 500, or 1,00 mg/kg, 1 month                                                      No effects on reproduction: NOAEL = 1 g/kg                                                                                                                                               

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Fucoidans           *Undaria pinnatifi*da                                       Rats                                                 Subchronic toxicity               1.35 g/kg, 1 month                                                                    No evidence of toxicity                                                                                                                                        91.8 g                    \[[@B152]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Galacto-mannans     *Cyamopsis tetragonolobus*                                  Adolescent and adult ♂ rats                          Subchronic and chronic toxicity   8% of diet, 6-67 weeks                                                                No evidence of toxicity                                                                                                                                        8% of diet                \[[@B153]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  Rats                                                 Acute toxicity                    One 7.06 g/kg dose: observed 2 weeks                                                  LD~50~= 7.06 g/kg                                                                                                                                              480 g                     \[[@B96]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                       Subchronic and chronic toxicity   1, 2, 4, 7.5 or 15% of diet, 3 months                                                 All doses ↓ ♀ BW; 7.5-15% ↓ ♂ BW; 15% ↓ bone marrow cellularity; ↓ kidney and liver weights                                                                    1-15% of diet             

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  19 adults with hypercholesterol-emia                                                   18 g/day, 1 year                                                                      Short-term gastric bloating/loose stools, in 8 subjects, resolved in 7-10 days; 2 withdrew because of diarrhea. No toxicity for 13 subjects completing study   18 g                      \[[@B154]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  16 Type II diabetics                                                                   26.4-39.6 g/day, 6 months                                                             No effects on hematologic, hepatic, or renal function                                                                                                          39.9 g                    \[[@B155]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  18 Type II diabetics                                                                   30 g/day, 4 months                                                                                                                                                                                                                                   30 g                      

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Cyamopsis tetragonolobus*(partially hydrolyzed guar gum)   Mice & rats                                          Acute toxicity                    One 6 g/kg dose; observed\                                                            LD~50~\> 6 g/kg                                                                                                                                                \>408 g                   \[[@B156]\]
                                                                                                                                                                         2 weeks                                                                                                                                                                                                                                                                        

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  Rats                                                 Subchronic toxicity               0.2, 1.0 or 5% of diet, 13 weeks                                                      No evidence of toxicity                                                                                                                                        5% of diet                

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                                                         0.5 or 2.5 g/kg, 1 month                                                              NOAEL \> 2.5 g/kg                                                                                                                                              \>170 g                   \[[@B157]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  *S. typhimurium*                                     Mutagenicity                      Ames test                                                                             Not mutagenic                                                                                                                                                  NA                        

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Glucans             *Agaricus subrufescens*(aqueous extract)                    Rats                                                 Subchronic toxicity               0.63, 1.25, 2.5 or 5% of diet, 3 months                                               NOAEL = 5% of diet                                                                                                                                             5% of diet                \[[@B158]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  3 women with advanced cancers                        Case reports                      Specific identity of products, doses, and durations of intake unknown                 Severe hepatotoxicity; two patients died                                                                                                                       NA                        \[[@B97]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Agaricus subrufescens*(freeze dried powder)                24 normal adults and 24 adults with liver problems   Subchronic toxicity               3 g, 4 months                                                                         No evidence of toxicity                                                                                                                                        3 g                       \[[@B159]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Ganoderma lucidum*\                                        Elderly woman                                        Case report                       1 year *G. lucidum*(and another unidentified product, initiated one month previous)   Elevated liver enzymes and liver tissue damage                                                                                                                 NA                        \[[@B98]\]
                      (supplement)                                                                                                                                                                                                                                                                                                                                                                                                                      

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Grifola frondosa*(powder)                                  Rats                                                 Acute toxicity                    One 2 g/kg dose                                                                       No evidence of toxicity                                                                                                                                        136 g                     \[[@B160]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Lentinula edodes*(powder)                                  10 adults                                            Safety                            4 g/day for 10 weeks; repeated\                                                       50% of subjects experienced blood eosinophilia, ↑ eosinophil granule proteins in serum and stool, ↑GI symptoms                                                 4 g                       \[[@B99]\]
                                                                                                                                                                         3-6 months later                                                                                                                                                                                                                                                               

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Lentinula edodes*\                                         Nude mice                                            Safety                            10% of diet days 1-18, 33-50                                                          No adverse events                                                                                                                                              10% of diet               \[[@B80]\]
                      (SME)                                                                                                                                                                                                                                                                                                                                                                                                                             

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  61 men with prostate cancer                                                            0.1 g/kg, 6 months                                                                    No adverse events                                                                                                                                              6.8 g                     

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Lentinus lepideus*(PG101)                                  Female mice                                          Subchronic toxicity               0.5 g/kg, 24 days                                                                     No evidence of toxicity                                                                                                                                        34 g                      \[[@B92]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Phellinus linteus*\                                        Rats                                                 Acute toxicity                    One 5 g/kg dose; observed\                                                            LD~50~\> 5 g/kg                                                                                                                                                349 g                     \[[@B161]\]
                      (crude extract)                                                                                                                                    2 weeks                                                                                                                                                                                                                                                                        

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Pleurotus ostreatus*(aqueous extract)                      Mice                                                 Acute toxicity                    One 3 g/kg dose; observed\                                                            LD~50~\> 3 g/kg                                                                                                                                                \>204.g                   \[[@B100]\]
                                                                                                                                                                         1 day                                                                                                                                                                                                                                                                          

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                       Subacute toxicity                 319 mg/kg, 1 month                                                                    Hemorrhages in intestine, liver, lung, kidney; inflammation and microabscesses in liver                                                                        21.7 g                    

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Saccharomyces cerevisiae*(particulate glucan \[WGP\])      Rats                                                 Acute toxicity                    One 2 g/kg, observed 2 weeks                                                          LD~50~\> 2 g/kg                                                                                                                                                \>136 g                   \[[@B144]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                                                                       Subchronic toxicity               2, 33.3 or 100 mg/kg, 3 months                                                        NOAEL = 100 mg/kg                                                                                                                                              6.80 g                    

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Heteroglycans       *Trametes versicolor*\                                      Rats                                                 Subchronic toxicity               1.5, 3.0 or 6.0 mg/kg, 2 months                                                       No evidence of toxicity                                                                                                                                        408 mg                    \[[@B162]\]
                      (PSP)                                                                                                                                                                                                                                                                                                                                                                                                                             

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  Rats & monkeys                                       Subchronic and chronic toxicity   100-200X equivalent human dose, 6 months                                              No evidence of toxicity                                                                                                                                        NA                        

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                      *Trametes versicolor*\                                      Humans with colon cancer                             Safety                            3 g/day, up to 7 years                                                                No significant adverse events                                                                                                                                  3 g                       \[[@B57]\]
                      (PSK)                                                                                                                                                                                                                                                                                                                                                                                                                             

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  Humans with colorectal cancer                                                          3 g/day, 2 years                                                                                                                                                                                                                                     3 g                       \[[@B58]\]

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

  Mannans             *Aloe vera*gel                                              Dogs                                                 Acute toxicity                    Fed one 32 g/kg; observed 2 weeks                                                     LD50 \> 32 g/kg                                                                                                                                                \>2,176 g                 Bill Pine, personal communi-cation

                                                                                                                                                                                                                                                                                                                                                                                                                                                        

                                                                                  Rats                                                                                   One 21.5 g/kg; observed 2 weeks                                                       LD50 \> 10 g/kg                                                                                                                                                \>680 g                   
  ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

\*150 lb adult

A number of studies in healthy human adults demonstrated immune stimulating effects of oral polysaccharides. Arabinogalactans from *Larix occidentalis*(Western larch) were shown in RCTs to increase lymphocyte proliferation and the number of CD8+ lymphocytes \[[@B18]\] and to increase the IgG subtype response to pneumococcal vaccination \[[@B19]\]. A furanose extract from *Panax quiquefolium*(North American ginseng) was shown in an RCT of healthy older adults to decrease the incidence of acute respiratory illness and symptom duration \[[@B20]\]. Finally, an RCT of healthy adults consuming *Undaria pinnatifida*(wakame) fucoidans found both immune stimulating and suppressing effects, including increased stromal-derived factor-1, IFN-g, CD34+ cells and CXCR4-expressing CD34+ cells and decreased blood leukocytes and lymphocytes \[[@B21]\].

Studies in healthy animals showed a number of immune stimulating effects of various glucan products from *Agaricus subrufescens (A. blazei)*(aqueous extracts \[[@B22]\], aqueous extracts with standardized β-glucans \[[@B23]\], α-1,6 and α-1,4 glucans \[[@B24]\], and whole plant powders \[[@B25]\]); *Lentinula edodes*(shiitake) (lentinan \[[@B26]\] and β-glucans \[[@B27]\]); *Saccharomyces cerevisiae*(β-1,3-glucans \[[@B27],[@B28]\]); *Laminaria digitata*(laminarin \[[@B29]\]); *Sclerotium rofsii*(glucan phosphate \[[@B29]\]); *Sclerotinia sclerotiorum*(SSG \[[@B30]\]); and *Phellinus linteus*(powder \[[@B31]\] and aqueous, alcohol-precipitated extract \[[@B32]\]). A furanose extract from *P. quiquefolium*and pectins from *Buplerum falcatum*and *Malus*(apple) spp. have also been shown to enhance immune function in healthy young animals \[[@B33]-[@B35]\]. *Cyamopsis tetragonolobus*galactomannan (guar gum) or highly methoxylated pectin feeding exerted numerous stimulating effects on antibody production in older animals \[[@B36]\].

Evidence for the effectiveness of oral polysaccharides against infection and immune challenges has been mainly demonstrated in animals. Immune stimulating effects have been shown in resting and exercise-stressed animals with thioglycollate, clodronate, or HSV-1 injections fed *Avena*(oat) spp. soluble glucans \[[@B37]-[@B41]\]; animals injected with or fed *E. vermiformis*and fed *Avena*spp. particulate glucans \[[@B42],[@B43]\]; animals with *E. coli*injections fed *L. digitata*glucans (laminarin) \[[@B44]\]; animals with HSV injections fed *U. pinnatifida*fucoidans \[[@B45]\]; animals with *Staphylococcus aureus*or *Candida albicans*injections fed *S. cerevisiae*glucans (scleroglucan) \[[@B29]\]; and animals with fecal solution injections fed an aqueous extract of *A. subrufescens*(*A. blazei*Murrill) \[[@B46]\].

Additional controlled human and animal studies have shown anti-inflammatory and anti-allergy effects of some polysaccharide products. In an RCT of adults with seasonal allergic rhinitis, *S. cerevisiae*β-1,3;1-6 glucans decreased IL-4, IL-5 and percent eosinophils, and increased IL-12 in nasal fluid \[[@B47]\], while a placebo-controlled study of patients with recurrent aphthous stomatitis (canker sores) consuming β-1,3;1-6 glucans found increased lymphocyte proliferation and decreased Ulcer Severity Scores \[[@B48]\].

Animal models of inflammatory bowel disease have shown anti-inflammatory effects of *Cladosiphon okamuranus*Tokida fucoidans \[[@B49]\], *Cyamopsis tetragonolobus*galactomannans \[[@B50]\], *Malus*spp. pectins \[[@B51]\], and mixed polysaccharide supplements \[[@B52]\]. Animals challenged with ovalbumin have demonstrated anti-inflammatory/allergy effects of *A. subrufescens*aqueous extracts \[[@B22]\], an aqueous extract *of Ganoderma tsugae*\[[@B53]\], and *Pyrus pyrifolia*pectins \[[@B54]\]. Anti-inflammatory effects have also been seen in animals with cotton pellet implantations fed a *Pholiota nameko*heteroglycan (PNPS-1) \[[@B55]\].

*Trametes versicolor*glucans have demonstrated anti-cancer effects in humans. In two RCTs and five controlled trials, PSK from *T. versicolor*mycelia increased survival of advanced stage gastric, colon and colorectal cancer patients \[[@B56]-[@B62]\] with one study showing increased immune parameters (including blood NK cell activity, leukocyte cytotoxicity, proportion of helper cells and lymphocyte suppressor cells) \[[@B62]\]. An RCT of advanced stage lung cancer patients consuming PSP from *T. versicolor*fruit bodies found increased IgG and IgM antibodies and total leukocyte and neutrophil counts, along with a decrease in the number of patients withdrawing from the study due to disease progression \[[@B63]\]. An RCT of ovarian or endometrial cancer patients consuming *A. subrufescens*glucans showed increased NK cell activity and fewer chemotherapy side effects \[[@B64]\].

In numerous animal models of cancer, a wide range of polysaccharides have shown anti-tumorogenic effects. Glucan products sourced from *A. subrufescens*demonstrating anti-cancer activities in animal models include an aqueous extract \[[@B65]\], an aqueous, acid-treated extract \[[@B66]\], and an aqueous extract with standardized levels of β-glucans \[[@B23]\]. Anti-cancer effects have been reported following intake of aqueous extracts of *G. lucidum*\[[@B67]-[@B69]\]; the powder and D fraction of *G. frondosa*\[[@B70]-[@B72]\]; *Hordeum vulgare*β-glucans \[[@B73]-[@B76]\]; *Laminaria angustata*powder \[[@B77]\]; *Lentinula edodes products*(powders \[[@B70],[@B78],[@B79]\], SME \[[@B80]\], β-glucans \[[@B27]\], and lentinan \[[@B81],[@B82]\]); *Pleurotus ostreatus*powder \[[@B70]\], *Saccharomyces cerevisiae*particulate β-1,3;1,6 and β-1,3glucans\[[@B27],[@B73]\]; and a glucan from *Sclerotinia sclerotiorum*(SSG) \[[@B30],[@B83]\]. A glucomannan from *L. edodes*(KS-2) improved survival of animals with cancer cell injections \[[@B84]\]; apple and citrus pectins have exerted anti-cancer effects, including decreased tumor incidence \[[@B85]-[@B90]\]. Finally, heteroglycans from *Lycium barbarum*(LBP~3p~), *Lentinus lepidus*(PG101) and A. *subrufescens*(ATOM) demonstrated a number of immune stimulating effects in animal cancer models \[[@B91]-[@B93]\]. Interestingly, only one animal study has been performed using glucans from *T. versicolor*(PSP): animals with cancer cell implantations showed decreased tumor growth and vascular density \[[@B94]\].

Most polysaccharide products appear to be safe, based on NOAEL, acute and/or chronic toxicity testing in rodents (Table [6](#T6){ref-type="table"}). As would be expected, powders, extracts and products that have not been fully characterized pose the most concerns. Other than for aloe vera gel, which was shown in a small human trial to increase the plasma bioavailability of vitamins C and E \[[@B95]\], the impact of polysaccharide intake on the absorption of nutrients and medications is not known. While one rat toxicity study raised concerns when guar gum comprised 15% of the daily diet \[[@B96]\], the product was safe in humans studies when 18-39.6 g/day was consumed for up to a year (Table [4](#T4){ref-type="table"}). Product contamination may explain three case reports of hepatotoxicity and/or death following intake of an *A. subrufescens*aqueous extract \[[@B97]\]. Seven animal studies reporting positive immunologic effects of *A. subrufescens*extracts in healthy animals or animals with cancers found no evidence of toxicity (Tables [1](#T1){ref-type="table"} and [2](#T2){ref-type="table"}). In humans, six weeks of *A. subrufescens*glucans intake was safe for cancer patients, and four months of 3 g/day intake by 24 healthy adults and 24 adults with liver disease reported no evidence of toxicity (Table [4](#T4){ref-type="table"}). Another case report associated liver toxicity with *G. lucidum*intake, but the elderly subject also took an unidentified product a month previous to her admission for testing \[[@B98]\]. Three animal studies reported immunologic benefits and no adverse effects following intake of *G. lucidum*aqueous extracts; in one study intake was 5% of the diet for 5 months (Table [1](#T1){ref-type="table"}). While adverse effects were also reported in a study in which 10 adults consumed 4 g/day *L. edodes*powder for 10 weeks \[[@B99]\], immunologic animal studies reported no ill effects of either *L. edodes*powder (5 studies, up to 5% of the diet up to nine months) or extract (7 studies, up to 40 days intake) (Tables [1](#T1){ref-type="table"} and [3](#T3){ref-type="table"}). Finally, while intake of 319 mg/kg of an aqueous extract of *P. ostreatus*by mice for 1 month caused hemorrhages in multiple tissues \[[@B100]\], there was no reported toxicity when mice consumed the mushroom powder as 5% of their diet for nine months (Table [3](#T3){ref-type="table"}). While ≥1 gram/day of *T. versicolor*glucan products were safely consumed by cancer patients for up to 10 years, the long-term effects of ingestion of the other polysaccharide products discussed in this review is also not known.

Discussion
==========

The majority of studies that qualified for inclusion in this review employed models investigating immune stimulation; fewer explored anti-inflammatory effects. Animal studies reported immune system effects in the gut, spleen, bone marrow, liver, blood, thymus, lungs, and saliva; controlled human studies reported evidence of immune stimulation in the blood, anti-inflammatory effects in nasal lavage fluid and improved survival in cancer patients. The literature is highly heterogenous and is not sufficient to support broad structure/function generalizations. For the limited number of studies that investigated well-characterized, isolated products (primarily glucan products), effects can be unequivocally attributed to polysaccharides. Such associations are certainly more tenuous when considering product powders or products obtained by extraction methods designed to isolate polysaccharides, but without complete compositional analyses.

Dietary polysaccharides are known to impact gut microbial ecology \[[@B101],[@B102]\], and advances in microbial ecology, immunology and metabolomics indicate that gut microbiota can impact host nutrition, immune modulation, resistance to pathogens, intestinal epithelial development and activity, and energy metabolism \[[@B103]-[@B107]\]. Other than fucoidans, the polysaccharides discussed in this review appear to be at least partially degraded by bacterial enzymes in the human digestive tract (Table [7](#T7){ref-type="table"}). Arabinogalactans, galactomannans, a glucan (laminarin), glucomannans, and mixed polysaccharide products (Ambrotose^®^products) have been shown to be metabolized by human colonic bacteria. Orally ingested fucoidans, glucans and mannans (or their fragments) have been detected in numerous tissues and organs throughout the body \[[@B73],[@B108],[@B109]\], (Carrington Laboratories, personal communication). We know of no study that has determined the specific identity of orally-ingested polysaccharide end products in animal or human tissues.

###### 

Fate of Immunomodulatory Polysaccharide Products Following Oral Intake

  --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
  Category                        Product                                                                           Metabol-ized by human gut bacteria?   Study type   Fate\                                                                                                               References
                                                                                                                                                                       (method: tissues detected)                                                                                          
  ------------------------------- --------------------------------------------------------------------------------- ------------------------------------- ------------ ------------------------------------------------------------------------------------------------------------------- ---------------------------------------------------
  Arabinogalactans                *Larix*spp.                                                                       yes                                   *in vitro*   NA                                                                                                                  \[[@B163]-[@B169]\]

                                                                                                                                                                                                                                                                                           

  Fucoidans                       *Undaria pinnatifida*                                                             no                                    *in vitro*   Ab: human plasma                                                                                                    \[[@B108],[@B170]\]

                                                                                                                                                                                                                                                                                           

  Galactomannans                  *Cyamopsis tetragonolobus*(partially hydrolyzed guar gum)                         yes                                   *in vivo*    NA                                                                                                                  \[[@B171]\]

                                                                                                                                                                                                                                                                                           

                                  *Cyamopsis tetragonolobus*(guar gum)                                              yes                                   *in vitro*   NA                                                                                                                  \[[@B167]\]

                                                                                                                                                                                                                                                                                           

  Glucans                         *Hordeum vulgare*                                                                 NA                                    *in vivo*    Fluorescein-labeled: mouse Mø in the spleen, bone marrow, lymph nodes                                               \[[@B73]\]

                                                                                                                                                                                                                                                                                           

                                  *Laminaria digitata*(laminarin)                                                   yes                                   *in vitro*   NA                                                                                                                  \[[@B29],[@B170],[@B172]\]

                                                                                                                                                                                                                                                                                           

                                  *Sclerotium rofsii*(scleroglucan) glucan phosphate, *Laminaria*spp. (laminarin)   NA                                    *in vivo*    Alexa Fluor 488-labeled: mouse intestinal epithelial cells, plasma, GALT                                            \[[@B29]\]

                                                                                                                                                                                                                                                                                           

                                  *Saccharomyces cervisiae*(particulate)                                            NA                                    *in vivo*    Fluorescein-labeled: mouse macrophage in the spleen, bone marrow, lymph nodes                                       \[[@B73]\]

                                                                                                                                                                                                                                                                                           

                                  *Trametes versicolor*\                                                            NA                                    *in vivo*    ^14^C-labeled: rat and rabbit serum; mouse GI tract, bone marrow, salivary glands, liver, brain, spleen, pancreas   \[[@B173]\]
                                  (PSK)                                                                                                                                                                                                                                                    

                                                                                                                                                                                                                                                                                           

  Mannans                         *Aloe barbadensis*(aloemannan)                                                    yes                                   *in vitro*   FITC-labeled: mouse, GI tract                                                                                       \[[@B121],[@B174]\]

                                                                                                                                                                                                                                                                                           

                                  *Aloe barbadensis*\                                                               yes                                   *in vitro*   NA                                                                                                                  \[[@B163]\]
                                  (gel powder)                                                                                                                                                                                                                                             

                                                                                                                                                                                                                                                                                           

                                  *Aloe barbadensis*(acemannan)                                                     NA                                    *in vivo*    ^14^C-labeled: dog systemic, particularly liver, bone marrow, gut, kidney, thymus, spleen                           (Carrington Laboratories, personal communication)

                                                                                                                                                                                                                                                                                           

  Mixed polysaccharide products   Ambrotose complex^®^, Advanced Ambrotose^®^powder                                 yes                                   *in vitro*   NA                                                                                                                  \[[@B163],[@B175]\]

                                                                                                                                                                                                                                                                                           

  Pectins                         NA                                                                                yes                                   *in vitro*   NA                                                                                                                  \[[@B165]-[@B167],[@B176]\]

                                                                                                                                                                                                                                                                                           

                                  *Buplerum falcatum*(bupleuran 2IIc)                                               NA                                    *in vivo*    Ab bound: mouse Peyer\'s patch, liver                                                                               \[[@B109]\]
  --------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

One can only speculate upon the mechanisms by which the polysaccharides discussed in this review influence immunologic function, particularly when one considers the exceedingly complex environment of the GI tract. It is possible that fragments of polysaccharides partially hydrolyzed by gut bacteria may either bind to gut epithelia and exert localized and/or systemic immune system effects, or be absorbed into the bloodstream, with the potential to exert systemic effects. Current studies investigating the link between the bioconversion of dietary polysaccharides, their bioavailability and their downstream effects on the host metabolism and physiology are utilizing metabolomic and metagenomic approaches that can detect and track diverse microbial metabolites from immunomodulatory polysaccharides \[[@B103]\]. These and other innovative approaches in the field of colonic fermentation are providing novel insights into gut microbial-human mutualism \[[@B110],[@B111]\], its impact on regulating human health and disease, and the importance of dietary modulation \[[@B112]-[@B115]\].

Additional RCTs of well-characterized products are needed to more completely understand the immunomodulatory effects and specific applications of oral polysaccharides. Such studies will need to better investigate the optimal timing and duration for polysaccharide ingestion. That is, should they be consumed continuously, before, at the time of, or after exposure to a pathogen or environmental insult? Only a few studies have actually investigated the impact of timing of polysaccharide intake to achieve optimal benefits. Daily feeding with some polysaccharides appears to result in tolerance (and diminished benefits); this has been demonstrated for some mushroom β-glucans \[[@B3],[@B26]\]. For those polysaccharides whose immunologic effects are dependent on their prebiotic activities, regular feeding would be presumed necessary.

Conclusions
===========

The dietary polysaccharides included in this review have been shown to elicit diverse immunomodulatory effects in animal tissues, including the blood, GI tract, and spleen. In controlled human trials, polysaccharide intake stimulated the immune system in the blood of healthy adults, dampened the allergic response to a respiratory inflammatory agent, and improved survival in cancer patients. Additional RCTs of well-characterized products are needed to more completely understand the immunomodulatory effects and specific applications of oral polysaccharides

List of abbreviations
=====================

♀: female; ♂: male; Ab: antibody; AIDS: autoimmune deficiency syndrome; AOM: azoxymethane; BBN: N-butyl-N\'-butanolnitrosamine; BLCL: Burkitt\'s Lymphoma Cell Line; BW: body weight; CBC: complete blood count; CD: cluster of differentiation; CFU: colony forming unit; ConA: concanavalin A; CXCR: CXC chemokine receptor; DMBA: 7,12-dimethylbenz*(a)*anthracene; DMH: N-N\'-dimethylhydrazine; DMN: dimethylhydrazine; DSS: dextran sulfate sodium; EBV: Epstein-Barr virus; GALT: gut-associated lymphoid tissue; GI: gastrointestinal; H~2~0~2:~hydrogen peroxide; HSV: herpes simplex virus; ICR: imprinting control region; ID: intradermal; IEL: intraepithelial lymphocytes; IFN-λ: interferon gamma; IG: intragastric; IgA: immunoglobulin A; IgE: immunoglobulin E; IgG: immunoglobulin G; IgM: immunoglobulin M; IL: interleukin; IMC: invasive micropapillary carcinoma; IN: intranasally; IP: intraperitoneal; IV: intravenous; LPS: lipopolysaccharide; Mø: macrophage; mAb: monoclonal antibody; 3-MCA: methylcholanthrene; MLN: mesenteric lymph nodes; MM-46 carcinoma: mouse mammary carcinoma; MW: molecular weight; NK: natural killer; NOAEL: no observable adverse effect level; OVA: ovalbumin; PBL: peripheral blood leukocytes; PBMC: peripheral blood mononuclear cells; PHA: phytohaemagglutinin; PMA: phorbol 12-myristate 13-acetate; PML: polymorphonuclear lymphocyte; RCT: randomized, controlled trial; RNA: ribonucleic acid; SC: subcutaneous; SD rats: Sprague Dawley; TCR: T cell receptor; TLR: toll like receptor; TNF-α: tumor necrosis factor alpha; UC: ulcerative colitis; WT: wild type.

Competing interests
===================

The authors are employees of the Research & Development Department at Mannatech, Incorporated, which sells two of the polysaccharide products (Ambrotose^®^powder and Advanced Ambrotose^®^powder) discussed in this review.

Authors\' contributions
=======================

JER and EDN conducted literature searches and wrote the manuscript. RAS provided technical guidance. All authors read and approved the final manuscript.

Acknowledgements
================

The authors would like to thank Barbara K. Kinsey, Ward Moore and Mrs. Jennifer Aponte for their assistance with the preparation of this manuscript, and Dr. Azita Alavi and Mrs. Christy Duncan for their editorial assistance.
