Fig. 1
Screening and expansion of tumor-reactive T cells in non-metastatic lymph nodes. a Lymphocytes (1 × 105/well) after making cell suspension were stimulated by autologous tumor cells (1 × 105/well) and IFNγ secretions were detected by ELISPOT assay as shown Pre. Lymphocytes co-culture with autologous tumor-cells for 2 weeks were rested without tumor-cells for 2 days. Subsequently, lymphocytes (1 × 105/well) were stimulated by autologous tumor-cells (1 × 105/well) and IFNγ secretions were detected by ELISPOT assay as shown Post. Samples of which spot number (lymphocytes + tumor cells) is over 15 and more 1.5 folds than the negative control (only lymphocytes) were considered as positive. Experiments were conducted in duplicate. b Absolute live cell numbers were counted before and 2 weeks after co-culture with autologous tumor-cells. A part of lymphocytes was stained with anti-CD3, CD4, CD8α, CD56 and CD19 antibody and their expressions were evaluated by flow cytometry. The number indicates CD8+ T cells after co-culture with autologous tumor cells from 1 × 106 total lymphocytes