Fig. 5: Expression of antigen by innate-like T cells is sufficient to mediate induction of tolerance in vivo.
a, GFP expression by thymic nonstromal populations in IL-17A–GFP mice (as in Fig. 3a; n = 5 mice). See also Extended Data Fig. 5a–c; MAIT, mucosal-associated invariant T. b, Reciprocal BM chimeras were established and immunized with GFP200–208 peptide as in Fig. 2c but using IL-17A–GFP donor and recipients (all mice were on a CB6F1 background). WT mice were used as an unimmunized control. Tetramer binding by CD8+CD44+ T cells, frequencies of CD8+CD44+tetramer+ cells from total CD8+ T cells and their absolute numbers are shown. Results were pooled from two independent experiments (WT → WT and IL-17A–GFP → WT groups) or from one experiment (the rest; n = 4 mice for nonimmunized, n = 8 for WT → WT, n = 11 for IL-17A–GFP → WT, n = 3 for WT → IL-17A–GFP and n = 3 for IL-17A–GFP → IL-17A–GFP). c, RTOCs with sorted Jedi DP thymocytes and thymic iNKT cells were established and analyzed as in Fig. 4b but using IL-17A–GFP iNKT cells (sorted as CD1d tetramer-binding cells (without gating on GFP); n = 2 RTOCs for WT iNKT cells and n = 3 for IL-17A–GFP iNKT cells). Representative results of three independent experiments are shown. d, Jedi-TCRβ mice were injected intrathymically (i.t.) with iNKT cells sorted from the thymi of IL-4–GFP mice (both donor and recipient mice were on a BALB/c background). Thymi of the injected mice were analyzed 7 days later. Uninjected Jedi-TCRβ mice were used as controls. Representative plots show frequencies of GFP-expressing cells, frequencies of H2-Kd GFP200–208 tetramer-binding Jedi thymocytes and expression of the indicated cell-surface markers by the latter cells. Quantification of the frequencies of all tetramer-binding cells of total thymocytes, CD8α+CD8β+ tetramer-binding cells of total thymocytes and CD8α+CD8β+ cells of tetramer-binding cells are shown. Results are pooled from three intrathymic injection experiments (n = 8 lobes for the uninjected group and n = 6 lobes for the IL-4–GFP group; 2 lobes from the uninjected group were not stained for CD8 expression). Data are presented as mean ± s.d. (*P < 0.05, ***P < 0.001 and ****P < 0.0001) and were analyzed by two-tailed Student’s t-test (d) or one-way ANOVA with a Holm–Sidak multiple comparisons test (b).
