Immunologic defect may leave some vulnerable to rare bacterialinfection ljiresearch frontiersin
)]; epitopes. Peptides were synthesized by A&A LLC . Peptides were pooled into peptide pools, re-lyophilized and reconstituted at a concentration of 1mg/ml or 0.7mg/ml .Venous blood was collected in heparin or EDTA containing blood bags or tubes and PBMCs were isolated by density gradient centrifugation using Ficoll-Hypaque according to the manufacturer’s instructions. Cells were resuspended in FBS containing 10% DMSO and cryopreserved in liquid nitrogen.
Cryopreserved PBMC were quickly thawed by incubating each cryovial at 37°C for 2 min, and cells transferred to 9ml of cold medium , supplemented with 5% human AB serum , 1% penicillin streptomycin , 1% glutamax and 20 U/ml benzonase nuclease . Cells were centrifuged and resuspended in medium to determine cell concentration and viability using trypan blue and a hematocytometer.
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