Fig. 6: Transient priming with GSK improves the in vivo persistence of OT1 cells.

Percentages of Ki-67⁺ (a) and Annexin V⁺ (b) cells in transferred OT1 TILs or endogenous CD8⁺ TILs (c) isolated from B16-OVA tumors and analyzed by flow cytometry on day 5 post transfer (n = 5 mice per group). d Experimental design for co-transfer of Veh- and GSK-primed OT1 cells to B16-OVA tumor-bearing mice and TIL analysis by flow cytometry. e Frequencies of Veh-primed CD45.1/CD45.2 and GSK-primed CD45.2 OT1 cells in B16-OVA tumors analyzed by flow cytometry on day 5 (n = 5 mice per group) and day 8 (n = 3 mice per group) after co-transfer. Flow cytometry analysis of percentages of Annexin V⁺ cells (f) and PD-1 expression (g) in OT1 TILs in the co-transfer assay (n = 3 mice per group). h Tumor weights of B16-WT tumor-bearing mice transferred with GSK- or Veh-primed OT1 cells (n = 5 mice per group). Frequencies of GSK- or Veh-primed OT1 cells in B16-WT tumors (i) and tumor-draining lymph nodes (TdLNs, j) analyzed by flow cytometry on day 5 post transfer (n = 5 mice per group). Frequencies (k) and phenotypes (l) of Veh-primed CD45.1/CD45.2 and GSK-primed CD45.2 OT1 cells in the spleens of mice immunized with OVA_257–264/poly(I:C) on day 1 and analyzed by flow cytometry on day 5 after co-transfer (n = 3 mice per group). m Frequencies of GSK- or Veh-primed OT1 cells in peripheral blood of OVA_257–264/poly(I:C)-immunized mice, analyzed by flow cytometry on day 14 and day 28 after co-transfer (n = 3 mice per group). Data in this figure are presented as mean ± SEM and represent three (a–c) or two (e–g, k–m) independent experiments. Statistical significance was determined by two-sided unpaired t-test (a–c, h–j) or paired t-test (e–g, k–m). Source data are provided as a Source Data file.