Extended Data Fig. 5: CD8⁺ T cell-specific SP immunization suppresses MOG_35–55-specific CD4⁺ T cells and induces CD8⁺ T cells with a regulatory phenotype.

a–c, C57BL/6J mice were immunized with an emulsion containing MOG_35–55, CFA and PTX (n = 5), or MOG_35–55, CFA, PTX and SP (n = 5). From unimmunized mice (a) and mice ten days after immunization (b, c) spleen and LN cells were isolated, stained and enriched for MOG_35–55 I-A^b pMHC-specific CD4⁺ T cells and an irrelevant tetramer. Representative FACS plots for different groups are shown. d–g, Spleen and LN cells were isolated from unimmunized mice (n = 5) (d) and mice 10 days after immunization with MOG (n = 5) (g), MOG plus SP (n = 5) (f) or SP alone (n = 4) (e), and then stained and enriched for SP-specific CD8⁺ T cells using the pMHC tetramer. Representative FACS dot plots for CD8⁺ T cell with a regulatory phenotype (CD44⁺CD122⁺Ly49⁺) from each group are shown. h, Tetramer-positive (that is, ASR, HDR, SMRP and YQP-tet⁺) CD8⁺ T cells were sub-gated for CD122, CD44 and Ly49, and the frequency of CD122⁺CD44⁺Ly49⁺ cells among SP-specific cells are shown among different immunization groups. ***P = 0.0002, one-way ANOVA followed by Tukey’s post hoc multiple comparison test. Data are mean ± s.e.m. and representative of two independent experiments.