Cd27 alexa fluor700

The following directly conjugated antibodies were used: CD3-APCH7, CD27-Alexa Fluor700, MIP1β-PE, CTLA4-APC, TNFα-Cy7PE, IL2-Cy55PCP (BD Biosciences); CD4-Cy55PE, Streptavidin (SA)-quantum dot (QD) 655, Aqua amine reactive dye (Invitrogen); CD45RO-TRPE, 2B4-Cy5PE (Beckman Coulter); IFNγ-eFluor450 (eBioscience); Biotinylated PD-1 (clone BAF1086; R&D Systems); CD8-BV711 (Biolegend). The following antibodies were conjugated in our laboratory: CD8-QD800 and CD57-QD565.

The following conjugated antibodies were used for cell-surface staining: CD3-Pacific Blue (BD Pharmingen; Clone UCHT1; Cat. No. 558117; San Diego, CA, USA), CD8-APC H7 (BD Pharmingen; Clone SK1; Cat. No. 641400), CD45RA-PE (BD Pharmingen; Clone HI100; Cat. No. 555489), CCR7-PE-Cy7 (BD Pharmingen; Clone 3D12; Cat. No. 557648), CD28-PerCP-Cy5.5 (BD Biosciences; Clone L293; Cat. No. 337181; San Jose, CA, USA), CD27-Alexa Fluor 700 (BD Pharmingen; Clone M-T271; Cat. No. 560611), CD95-APC (BD Pharmingen; Clone DX2; Cat. No. 558814) and CD127-FITC (BD Pharmingen; Clone HIL-7R-M21; Cat. No. 560549). Conjugated antibodies for intracellular staining included the following: IFN-γ-FITC (BD Pharmingen; Clone 4S.B3; Cat. No. 554551), IL-2-PerCP-Cy5.5 (BD Pharmingen; Clone MQ1-17H12; Cat. No. 560708) and TNF-α-AlexaFluor 700 (BD Pharmingen; Clone MAb11; Cat. No. 557996). To exclude dead cells, the Fixable Aqua Dead Cell Stain viability marker was used (Invitrogen; Cat. No. L34957; Eugene, OR, USA).