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. 2020 Aug 6;16(8):e1008696. doi: 10.1371/journal.ppat.1008696

Fig 5. Differential expression of co-inhibitory receptors by CD8+ T cells upon recognition of their cognate epitopes in vitro.

Fig 5

(A) Representative plots showing gating strategy for identification of antigen-specific T cells using tetramer staining. (B, C) Representative flow cytometry plot showing % co-expression of Tetramer+PD-1+ CD8+ T cells following stimulation of PBMCs with B27-KK10 (KRWIILGLNK) epitope (2 μg/ml) for 72 hrs in patient #LNP01 and patients #LNP03 respectively. (D, E) Representative flow cytometry plot showing % co-expression of Tetramer+PD-1+ CD8+ T cells following stimulation of PBMCs with A03-RK9 (RLRPGGKKK) and B35-FL9 (FPVKPQVPL) epitopes (2 μg/ml) for 72 hrs in patient #PR03 and patients #PR01 respectively. (F) Cumulative data showing % tetramer+PD-1+ in CD8+ T cells restricted by different HLA-alleles, as shown, following encounter with their cognate epitopes for 72 hrs. (G) Representative flow cytometry plot showing % co-expression of Tetramer+TIM-3+ CD8+ T cells following stimulation of PBMCs with A02-SL9 (SLYNTVATL) and B35-FL9 (FPVKPQVPL) epitopes (2 μg/ml) for 72 hrs in patients #PR04 and #PR01, respectively. (H) Cumulative data showing % TIM-3 expressing antigen-specific CD8+ T cells following stimulation with their cognate epitopes for 72 hrs. (I, J) Representative flow cytometry plot showing % co-expression of Tetramer+CTLA-4+ CD8+ T cells following stimulation of PBMCs with B35-FL9 (FPVKPQVPL), A03-RK9 (RLRPGGKKK), and B57-KF-11 (KAFSPEVIPMF) epitopes (2 μg/ml) for 72 hrs in patients #PR01 and #PR03 and #LNP02, respectively. (K) Cumulative data showing % TIM-3 expressing antigen-specific CD8+ T cells following stimulation with their cognate epitopes for 72 hrs. Each point represents data from an epitope. Bar, mean ± one standard error.