IVS assays were performed as described previously (8 (link)). Briefly, PBMCs were stimulated for 6 d in the presence of 10 μg/ml anti-IL-10R and/or anti-PD-1 (clone EH12.2H7, Biolegend) blocking mAb or isotype control antibodies prior to intracellular cytokine staining. Alternatively, cells were harvested and then surface stained with APC-labeled A2/NY-ESO-1 tetramers, and subsequently with CD8-PECy7 and Annexin V-FITC (ApoScreen Annexin V-FITC Apoptosis Kit, Beckman Coulter).
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IVS Assays for Immune Checkpoint Inhibition
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Other organizations : University of Pittsburgh
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Variable analysis
independent variables
- Anti-IL-10R blocking mAb
- Anti-PD-1 (clone EH12.2H7, Biolegend) blocking mAb
- Intracellular cytokine staining
- Surface staining with APC-labeled A2/NY-ESO-1 tetramers
- CD8-PECy7 staining
- Annexin V-FITC staining
- Isotype control antibodies
- Positive control: None specified
- Negative control: Isotype control antibodies
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