recently , a single center study conducted by oiva and coworkers and published in critical care   demonstrated that phospho - specific whole blood flow cytometry could be used to assess activated signaling path - ways in leukocytes isolated from pancreatitis patients .
pancreatitis is an inflammatory process in which pancreatic enzymes are activated leading to autodigestion of the pancreas and adjacent tissues . during pernicious cases ,
the initial inflammatory response is excessive and can result in tissue injury , organ failure , and death . to counter this inflammation ,
these steps can include leukocyte apoptosis , anti - inflammatory cytokine production , and anergy .
when patients become leukopenic and immunosuppressed , they often become susceptible to infections with organisms to which a healthy patient would normally be resistant .
although therapy aimed at blunting the initial over - exuberant inflammation may be beneficial if applied early , it may also be detrimental by impairing the host 's ability to fight infection that is a common cause of death in these patients . to complicate the potential use of immune - modulating therapy ,
inflammatory and anti - inflammatory processes can occur along different time - lines for the varied leukocyte subsets .
for example , during immune response , t lymphocytes might be undergoing apoptosis while neutrophils are primed to produce reactive oxygen species .
thus , during prolonged pancreatitis , leukocyte subsets may be concurrently exhibiting both pro- and anti - inflammatory processes .
as noted above , a potential promising therapy of pancreatitis is modulation of the immune response .
this is a difficult undertaking as the host response will be affected by a number of variables , including genetic background , co - morbidities , age , gender , and so on .
in addition , the patient 's immune response will vary during the course of the disease .
thus , what is needed for immunotherapy to be practical is a rapid , robust measure of the host immune system . in the manuscript published by oiva and co - workers
, the authors determined the signaling profiles of circulating leukocytes isolated from pancreatitis patients using phospho - specific whole blood flow cytometry .
this is a powerful new technology that allows for simultaneous single - cell determination of leukocyte subsets using cell surface markers as well as intracellular protein phosphorylation .
this work represents a continuation of work published previously . in this earlier work , oiva and co - workers
demonstrated that stimulated monocytes isolated from patients with acute pancreatitis had decreased phosphorylated erk 1/2 , nf-b , and stat1 and 3 .
the authors concluded that these changes could lead to impaired monocyte recruitment as well as increased susceptibility to infections .
here , changes in activated t lymphocyte p38 , nf-b , stat1 and stat6 activity was observed that could be interpreted as being pro- or anti - inflammatory .
interestingly , they determined that patients ' lymphocytes exhibited decreased phosphorylated nf-b and stat1 and increased phosphorylation of p38 and stat6 , suggesting a transition from a th1 to th2 phenotype .
additionally , this minimally invasive methodology could be used to generate immune status within 6 hours under ideal conditions .
thus , this methodology represents an essential step prior to targeting the underlying immune response to pancreatitis .
although beyond the scope of the report , a potentially necessary step after immune status determination , and prior to treatment with inflammatory altering treatments , would be to first treat the whole blood cells isolated from the patients with the potential therapeutic agent and evaluate the immune effector cell response .
phospho - specific flow cytometry could also be used to determine if the leukocytes responded in a way that would be beneficial .
thus , the minimally invasive , relatively quick methodology developed in this paper could be utilized to determine immune status as well as provide a method to test potential therapies .
one unavoidable limitation to the report is that the signaling processes were determined using only peripheral leukocytes .
a possibility exists that peripheral leukocytes may not respond similarly to ex vivo stimuli as leukocytes isolated from the inflammatory site(s ) . when feasible , future studies need to be undertaken to compare the response of peripheral leukocytes to these tissue leukocytes .
rapid results from whole blood phospho - specific flow cytometry during pancreatitis will allow for immune status determination , likely improve early diagnosis and provide a rational basis for immune targeting therapies .
this work was supported in part by national institutes of health grants gm44118 and gm55194 ( to rsh ) , gm72760 ( to ccc ) , and the alan a and edith l wolff foundation .
 recently , a single center study conducted by oiva and coworkers and published in critical care demonstrated that phospho - specific whole blood flow cytometry could be used to assess activated signaling pathways in leukocytes isolated from pancreatitis patients . 
 the authors demonstrated that this methodology had the potential to determine the current status of a patient 's immune state . 
 although the experimental cohort was clinically homogeneous , the observed data were heterogeneous . 
 altogether , these results suggest that prior to administering immune - modulatory therapies in inflammatory diseases , it will be beneficial to first determine immune status 
 . rapid results from whole blood phospho - specific flow cytometry may allow for determination of immune status , improve early diagnosis , and provide a rational basis for immunomodulatory therapies . 
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