Anti ccr7apc

Activation-induced markers were quantified via flow cytometry (FACSLyric, BD Biosciences). A surface staining on PBMC was performed with anti-CD3^PerCP (BD, clone SK7), anti-CD4^V450 (BD, clone L200), anti-CD8^FITC (DAKO, clone DK25), anti-CD45RA^PE-Cy7 (BD, clone L48), anti-CCR7^APC (R&D Systems, clone 150503), anti-CD69^APC-H7 (BD, clone FN50) and anti-CD137^PE (Miltenyi, clone 4B4-1). T-cell subsets were identified via the following gating strategy: LIVE CD3⁺ were selected and divided in CD3⁺CD4⁺ and CD3⁺CD8⁺. Within the CD4 and CD8 subsets, memory subsets were gated as CD45RA⁺CCR7⁺ (naive, T_N), CD45RA^-, CCR7⁺ (central memory, T_CM), CD45RA^-CCR7^- (effector memory, T_EM) or CD45RA⁺CCR7^- (terminally differentiated effectors, T_EMRA). T cells specifically activated by SARS-CoV-2 were identified by up-regulation of CD69 and CD137. An average of 500,000 cells was always acquired, the gating strategy is schematically represented in (Fig. S1A-J). In analysis, PBMC stimulated with MP_CD8_A and MP_CD8_B were concatenated and analyzed as a single file for SARS-CoV-2-specific responses to MP_CD8.

Flow cytometric analysis of surface marker expression was performed on a FACS Canto II flow cytometer (Becton Dickinson) using a five-color setup. Anti CD206 FITC, Anti CD163 PE and Anti CD14 PerCP were bought from Becton-Dickinson. Anti-CCR7 APC was supplied by R&D systems and Anti CD11c PC7 by IOT/Beckman-Coulter.

Anti-CD3-PE-cy7, anti-CD45RA-FITC, anti-CD3-FITC/CD8-PE/CD45-PerCP/CD4-APC, and anti-CD38-PE reagents were purchased from BD Biosciences (San Jose, CA). Anti-CD4-APC-cy7, anti-CCR7-PerCP-cy5.5, anti-human leukocyte antigen DR-APC, and anti-IFN-γ-APC reagents were purchased from BD PharMingen (San Diego, CA). Anti-CD31-APC and anti-CD57-PE were purchased from BioLegend (San Diego, CA). Anti-CCR7-APC was purchased from R&D Systems (Minneapolis, MN).