Extended Data Figure 9. Immunogenic chemotherapy supports adoptive T cell transfer only in the absence of B cells.

a, The experimental scheme. Immunogenic TRAMP-C2 cells were s.c. inoculated into WT or Tcrβ^-/- mice. After 30 days, the mice were divided into 4 groups (n=4-5/group): 1) control, 2) oxaliplatin (weekly), 3) ATCT, 4) ATCT plus oxaliplatin (weekly). The first oxaliplatin cycle was given at day 31. Two days after the second cycle, CD8⁺ T cells from CD45.1×CD45.2 WT mice (3 × 10⁶ cells) were transferred into tumor-bearing mice and this was followed by two more oxaliplatin cycles after which mice were sacrificed for analysis on day 59. b, Tumor volumes (mm³) c,d, Flow cytometric analysis of spleen (c) and tumor (d) cells after staining with CD45.1, CD45.2, CD8 and TCRαβ antibodies, confirming expansion of adoptively transferred T cells. e, tumor growth curves. f, The experimental scheme. Immunogenic TRAMP-C2 cells were s.c. inoculated into WT or Rag1^-/- × OT-1 mice (no B cells), that harbor CD8⁺ T cells specific for chicken ovalbumin which is not expressed by TRAMP-C2 cells. After 30 days, tumor-bearing Rag1^-/- × OT-1 mice were divided into 4 groups (n=3-4 mice per group): 1) control, 2) oxaliplatin treatment, 3) ATCT, 4) oxaliplatin treatment plus ATCT. The first oxaliplatin cycle was given at day 31. Two days after the second oxaliplatin cycle, CD8⁺ T cells (3 × 10⁶) from CD45.1×CD45.2 mice were adoptively transferred into tumor-bearing mice, which were sacrificed on day 59 and analyzed. g, Flow cytometric analysisof tumor-infiltrating cells stained with CD45.1, CD45.2, CD8 and TCRαβ antibodies, confirming infiltration of adoptively transferred T cells. h, Flow cytometric analysis of GrzB expression in adoptively transferred, tumor-infiltrating, CD8⁺ effector cells (CD45.1⁺CD8⁺CD44⁺) from tumor-bearing mice treated as above. i, Tumor volumes (mm³). j, tumor growth curves. k, The experimental scheme for Fig. 5a-f. Sixteen weeks old TRAMP;Rag1^-/- mice (no B and T cells) were treated with oxaliplatin (weekly). One day after the 1^st treatment cycle, CFSE-labeled splenocytes from either WT (B and T cells, SP-WT) or Jh^-/- (T but no B cells, SP-Jh^-/-) mice were transferred into the tumor-bearing mice (5 × 10⁶ T cells per mouse; 4-5 mice per group). l, m, After 6 days, one mouse from each group was sacrificed, and the proliferation of CD8⁺ (l) and CD4⁺ (m) T cells in bone marrow (BM), spleen and prostates was analyzed by CFSE staining and flow cytometry. n-r, After 3 more oxaliplatin cycles (4 weeks in total), the mice were sacrificed and analyzed. n, Frequency of adoptively transferred CD19⁺ cells amongst CD45⁺ cells in spleens and prostates 30 days after ACT. o, Flow cytometric analyses of CD19⁺ B lymphocytes for TIM-1 expression in spleens (left) and prostates (right) of abovemice. p-r, Flow cytometric analyses of T lymphocytes. Percentages of CD8⁺ and CD4⁺ T cells in LN (p); spleens (q); prostates (r) of above TRAMP;Rag1^-/- mice. Red:splenocytes from WT mice (T and B cell transfer), blue: splenocytes from Jh^-/- mice (T cell transfer). Results are means ± s.e.m. Mann-Whitney and t tests were used to calculate statistical significance.