Extended Data Fig. 4. Immunological characteristics of genetically engineered mice with multiple myeloma.

a) Stage of CD8⁺ T cells (a) and CD4⁺ T cells (b) during MGUS (n = 8) and MM (n = 27) progression in mice. Controls corresponded to YFP_cγ1 mice (n = 3). Phenotype of exhaustion markers in CD8⁺ T cells (a) and in CD4⁺ T cells (b) during MGUS (n = 24) and MM (n = 43) compared with control age-matched mice (n = 17) (complementary to Fig. 4a). Mean ±s.d. are represented. c) Percentage of NK cells with TIGIT and LAG3 expression in BM in YFP_cγ1 control mice (n = 4) and in mice with MGUS (n = 6) and MM (n = 26) (complementary to Fig. 4a). d) Tumors with higher number of immune cells (n = 27-34) in the BM contained an increased number of tumor-reactive CD8⁺ T cells that expressed TIGIT, and LAG3, in contrast to those cases with lower immune infiltrates (n = 22-25) (complementary to Fig. 4d). e) Immunohistochemical studies using antibodies to detect GFP⁺ transgenic MM cells or CD3⁺ T lymphocytes in BM sections from YFP_cγ1 control mice, BI_cγ1 mice and MI_cγ1 mice. f) Representation of the percentage (%) of the area in a region of interest in the BM that is occupied by CD3⁺ T cells with respect to non-GFP⁺ MM cells. Samples from YFP_cγ1 control mice (n = 2), BI_cγ1 mice (n = 2) and MI_cγ1 mice (n = 2) were included. g) Pearson correlation analyses between the percentages of NK cells in the mouse BM and those of T_reg cells in the BM (complementary to Fig. 4d). h) In MM patients, tumors with more abundant infiltrating immune cells (n = 31) contained an increased number of tumor-reactive PD-1⁺ CD4⁺ T cells and NK cells in the BM compared with MM cases with lower number of immune cells (n = 69) (complementary to Fig. 4g). i) Pearson correlation analyses between the percentages of NK cells in the BM of MM patients and those of T_reg cells in the BM (complementary to Fig. 4h). Boxes represent median, upper and lower quartiles and whiskers represent minimum to maximum range (d and h). Two-tailed Mann-Whitney test P values (d and h) are indicated. **p < 0.01; ***p < 0.001.