Rapid generation of multivirus-specific T lymphocytes for the prevention and treatment of respiratory viral infections

摘要

Multi-R-VSTs were surface-stained with monoclonal antibodies to: CD3, CD25, CD28, CD45RO, CD279 (PD-1) [Becton Dickinson (BD), Franklin Lakes, NJ], CD4, CD8, CD16, CD62L, CD69 (Beckman Coulter, Brea, CA) and CD366 (TIM-3) (BioLegend, San Diego, CA). Cells were pelleted in phosphate-buffered saline (PBS) (Sigma-Aldrich), then antibodies added in saturating amounts (5μl) followed by incubation for 15mins at 4°C. Subsequently, cells were washed, resuspended in 300μl of PBS and at least 20,000 live cells acquired on a Gallios? Flow Cytometer and analyzed with Kaluza? Flow Analysis Software (Beckman Coulter).