Figure 1.

Checkpoint blockade does not consistently increase cytokine production by TILs in short-term assays. TILs from patients LAU50 (a) or LAU1015 (b) were pre-incubated for one hour with the indicated antibodies (Abs) and incubated with T2 cells loaded with Melan-A/MART-1 peptides ELAGIGILTV (“ELA”; analog) or EAAGIGILTV (“EAA”; wildtype) for 5 hours in the presence of Brefeldin and of the indicated blocking antibodies. T cells were labeled with ELA-multimers and anti–IFNγ and anti-TNFα Abs. Melan-A specific cells within each patient’s cells (left panel) as well as representative percentages of IFNγ+ and TNFα+ within multimer+ cells are shown. (c-d) TILs from 5 patients (LAU50, LAU1015, LAU1660, 0MM7, 0DJP) were pre-incubated for one hour with the indicated mAbs and were co-cultured with their respective autologous melanoma cell lines for 5 hours in the presence of Brefeldin before intracellular labeling with anti–IFNγ and anti-TNFα Abs. (c) IFNγ production from one representative patient sample in multimer+ and multimer- CD8 T cells (LAU1015) is shown. (d) Average of at least two independent experiments per patient sample is shown.