Activation Induced marker assay was performed as reported earlier3 (link). Briefly, PBMCs (0.7 million cells/100 µl) were plated onto separate 96 well U bottom plates and stimulated with the cocktail of SARS-CoV-2 peptide pool (1 µg/mL). After overnight stimulation, cells were washed and stained 40 minutes with antibody cocktail containing the following fluorescently conjugated antibodies obtained from Biolegend, USA: CD3-APC-A750 (Cat. 300470), CD4-PB450 (Cat. 300521), CD8a-APC-A700 (Cat. 301028), OX40-APC (Cat. 350008), CD137-ECD (Cat. 309826), CD69-PC7 (Cat. 310912), CCR7-PE (Cat. 353204), CD45RA-FITC (Cat. 304148). The plates were centrifuged and the resuspended cells were labelled again with 7AAD solution (Cat. 6604104, Beckman) and analysed in a flow cytometer (CytoFlex S, Beckman Coulter).
Other organizations :
All India Institute of Medical Sciences, Pandit Bhagwat Dayal Sharma Post Graduate Institute of Medical Sciences, Nizam's Institute of Medical Sciences, Washington University in St. Louis, Rajarajeswari Medical College and Hospital, Indian Institute of Management Visakhapatnam, Narayana Health, Oyster & Pearl Hospital, Bhabha Hospital, Bharat Biotech (India)
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