Figure 4.

Exhausted CD8 T cells and Tregs are the most susceptible to intercellular transfer. (A, B) ProjecTILs analysis of T cell scRNA-seq data. (A) Projection of total, ZsG- and ZsG+ T cell scRNA-seq data on the reference atlas with pie charts showing proportions of each TIL subset. (B) Fold-change in the frequency of ZsG+ T cells compared to that of ZsG- T cells for each TIL subtype. (C) Representative flow cytometry analysis showing ZsG transfer into CD4 conventional TILs compared to Tregs from B16^ZsG tumors with summary graphs of ZsG MFI and frequency of these populations. (D) Violin plots showing expression level of Ikzf2 in ZsG- and ZsG+ Tregs (scRNA-seq data) with representative flow cytometry analysis demonstrating a higher percentage of Helios+Foxp3+ cells among ZsG+ CD4 TILs compared to ZsG- CD4 TILs from B16^ZsG tumors. (E) Violin plots showing expression level of Tbx21 in ZsG- and ZsG+ Tregs (scRNA-seq data) with representative flow cytometry analysis demonstrating a higher percentage of Tbet+Foxp3+ cells among ZsG+ CD4 TILs compared to ZsG- CD4 TILs from B16^ZsG tumors. (F) Representative flow cytometry analysis showing Tim3 and PD-1 expression in ZsG- and ZsG+ CD8 TILs from B16^ZsG tumors with summary graphs of Tim3 and PD-1 MFI and frequency of the Tim3+PD-1+ population presented as fold changes relative to CD8 TILs from B16 control tumors. (C, F) Mean ± s.e.m, paired T-test (C), Mann-Whitney U test (F): **p<0.01, ***p<0.001, ****p<0.0001. Data represent three independent experiments (C–F) and are pooled from at least three independent experiments (C, F).