Sample collection and handling
LabOne
RIND, FRANKLIN, STERILE, SECONDARY, CAVITY
All samples were collected at the study site, logged, and tracked (Saint Louis University Center for Vaccine Development in St. Louis, MO). Safety samples were shipped the same day as a collection to Quest Diagnostics, formerly LabOne (Lenexa, KS), for processing. Immunology samples for secondary and exploratory endpoints were processed at Saint Louis University. Serum and PBMC samples from each participant were stored here until completion of the study, at which point samples were shipped to Advanced Bioscience Laboratories (ABL Inc.; Rockville, MD) for assays and analysis in two separate shipments to mitigate the risk of sample loss due to temperature excursions. PBMCs were shipped using liquid nitrogen-charged dry shippers, and serum samples were shipped on dry ice.For the Mycobacteria Growth Indicator Tube (MGIT) assay, blood was collected using sterile sodium heparin collection tubes. For safety hematology tests, blood was collected in tubes with EDTA. To obtain serum samples for safety chemistry and antibody analysis, blood was collected using serum separator tubes (SST). For antibody analysis, following centrifugation at 750–930 × To obtain PBMC samples, blood was collected into 8-mL CPT tubes (BD Biosciences, Franklin Lakes, NJ). Following centrifugation at 1800 × For nasal sample collection, the same sterile PurFlock swab (Puritan Medical Products, Guilford, ME) was inserted into each nasal cavity alongside the nasal septum and rotated 360° in each direction waiting for 5 s after each rotation before carefully removing the swab. After sampling both nostrils, the swab tip was placed in 3 mL of buffer containing Dulbecco’s PBS (DPBS) with 5 mM Tear samples were collected by first squeezing an orange rind directly over the participant’s eye. The participant then blinked several times until tears were generated. The tears (target volume 125 µL) were collected using a capillary tube which drained into a screw-cap collection microtube. The affected eye was then irrigated with an eye wash solution and a commercial eye drop solution. Tear samples were held on wet ice for up to 2 h before processing by centrifugation at 20,000 × 