Fig. 3: Cbir1, MHC-II restricted, microbe-reactive TCR transgenic develop an innate-like phenotype in the gut.

LPLs from Cbir1 TCR transgenic line crossed to Rag−/− mice were analyzed directly ex vivo or stimulated in vitro. A Flow cytometric analysis of TCRβ + lin− cells from the colon lamina propria of Cbir1 Rag−/− mice stained for CD4 and CD8a. (left) and IL-7R and IL-18R stain of the gated CD4 (green) and CD4/CD8 double negative (DN, blue) cell populations (right). FACS plots are representative of experiments n = 5, representative of two independent experiments. B Quantification of flow cytometry data showing the percent of the TCRβ + lin− population that are CD4, CD8a, and DN (left), and the percent of each population that have the IL-18R/IL-7R expressing phenotype (right). Each point represents an individual animal. n = 5, representative of two independent experiments. Mean with standard deviation shown, one-way ANOVA with Tukey’s multiple comparison test. C Quantification of FACS data of the percent of T cells with a DN phenotype from Cbir1 colon LPL, lung, mesenteric lymph node (MLN), spleen, and thymus. Each point represents an individual mouse. n = 4, representative of two independent experiments. Mean with standard deviation shown, one-way ANOVA with Dunnett’s multiple comparisons test comparing each mean to colon. D Representative flow cytometry data (left) and quantification (right) of IL-7R and PLZF expression within the CD4 and DN T cell subsets with fluorescence minus one (FMO) control (right). E Representative flow cytometry histograms of sorted CD4 or DN splenic Cbir1 Rag−/− T cells labeled with CFSE and incubated for 5 days with peptide-pulsed bone marrow-derived DCs (BMDCs). Plots are representative of three independent experiments. F Representative flow cytometry histograms of sorted DN T cells from Cbir Rag−/− mice stained with CFSE and incubated for five days with BMDCs fed feces of steady-state or vancomycin (vanc) treated mice from the same animal facility. Plots are representative of two independent experiments. Source data are provided as a Source Data file.