Cd69 pc7

Activation Induced marker assay was performed as reported earlier^{3 (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).

Activation Induced marker assay was performed as reported earlier³⁰ . 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 min with antibody cocktail containing the following fluorescently conjugated antibodies obtained from Biolegend, USA: CD3-APC-A750 (300470), CD4-PB450 (300521), CD8a-APC-A700 (301028), OX40-APC (350008), CD137-ECD (309826), CD69-PC7 (310912), CCR7-PE (353204), CD45RA-FITC (304148). The plates were centrifuged and the resuspended cells were labelled again with 7AAD solution (6604104, Beckman) and analysed in a flow cytometer (CytoFlex S, Beckman Coulter).