Anti cd107a pacific blue

Following in vitro re-stimulation, cells were surface-stained with fluorochrome-labeled antibodies anti-CD3-Alexa-Fluor700, anti-CD8-V500 and anti-CD4-APC Cy7 (BD Biosciences, BioLegend). In addition, HLA class I CMV tetramers-PE labeled (A*01 VTE, A*02 NLV, B*07 TPR, B*08 ELR and B*035 IPS) (Beckman Coulter or IBA Solution for Life Sciences), and Live/Dead Fixable Blue Dead-Cell Stain (Invitrogen) was used for gating on viable cells. In some experiments we used surface-stained with fluorochrome-labeled antibodies anti-PD1-FITC and anti CD160-PE Cy7 (BD Biosciences, BioLegend). Cytofix/Cytoperm (BD Biosciences) reagents were used to fix and permeabilize cells for intracellular cytokine staining (ICS) using anti-IFN-γ-PE Cy7 (or BV605), anti-IL-2-FITC, anti-Granzyme B-PE CF 594, anti-CD107a-Pacific Blue, anti-T-bet-PE or BV655 (BD Biosciences) and anti-Eomes-Alexa-Fluor 660 (eBiosciences).

Following in vitro re-stimulation, cells were surface-stained with fluorochrome-labeled antibodies anti-CD3-Alexa-Fluor700, anti-CD8-V500 and anti-CD4-APC Cy7 (BD Biosciences) and Live/Dead Fixable Blue Dead-Cell Stain (Invitrogen) was used for gating on viable cells. Cytofix/ Cytoperm (BD Biosciences) reagents were used to fix and permeabilize cells for ICS using anti-IFN-γ-BV605, anti-TNF-α-PE-Cy7, anti-IL-2-BV650, anti-Granzyme B-PE CF 594, anti-CD107a-Pacific Blue and anti-T-bet-PE (BD Biosciences).