Fig 2: Expression of differentially expressed genes in MBCs.(a) Heatmap of select genes from RNA-seq analysis of MBCs (B220+IgDloGL7-CD38+S1pr2-Tomato+) from Hhexflox/+ (Het) and Hhexflox/flox (KO) S1pr2-ERT2-cre-TdTomato mice and GC B cells (B220+IgDloGL7+CD95+Ephrinb1+S1pr2Venus/+) 16 at day 11 post LCMV infection. Data are from 3 independent experiments with at least 3 mice per experiment pooled for each sample. (b) Expression of Bcl6 (left), S1pr2 (middle), and Bcl2 (right) in Hhex Het and KO GC B cells (B220+IgDloGL7+CD95+S1pr2-Tomato+) at day 12 post LCMV infection. Data are from 1 independent experiments with 4 mice. (c) Expression of Bcl6, Hhex, Tle3, and Bcl2 projected onto tSNE plot of Mem cluster (n=1,255 cells). Color scaled for each gene with log-normalized expression level noted. Cells for scRNA-seq sort were pooled from 4 mice.

Fig 3: The role of extrinsic MyD88 for effector CD4+ T cell survival. (A-C) Mice received 106 purified CD4+ T cells from naïve 1807 mice and were vaccinated with heat-killed yeast. (A) 1807 T cell contraction. At serial time intervals post-vaccination, the numbers of activated 1807 T cells were enumerated from the sdLN and spleen by FACS. * P < 0.05 vs. wild type control recipient mice. (B) Caspase 3 expression. Numbers in the dot plot indicate the numbers (top line) and frequencies (2nd line from top) of intracellular caspase 3 positive 1807 T cells within the CD4+ T cell gate at day 12 post-vaccination. * P < 0.05 vs. wild type control recipient mice. (C) The mean fluorescence intensity of Bcl2 and Bcl-xL expression in 1807 T cells transferred into wild type and Myd88 -/- recipients. Isotype controls are shown on the left of the graphs (unlabeled). (D) Adoptive transfer of naïve Bim-/- 1807 T cells and Fas-/- 1807 T cells into Myd88 -/- and wild type controls prior to vaccination. At days 7 and 35 post-vaccination, the numbers of adoptively transferred 1807 T cells were enumerated in the sdLN and spleen. Data are the mean ± SEM (n = 4–6 mice/group). Data are representative of two independent experiments. * P < 0.05 vs. corresponding wild type recipient mice. ** P < 0.05 vs. corresponding wild type 1807 cells.

Fig 4: MAIT cell alterations during obesity.a Representative dot plots of MAIT cell and control MR1-Tet staining. The numbers represent MAIT cells frequency among aßT cells. b MAIT cell frequency among aßT lymphocytes in blood (ND n = 6; HFD n = 6), liver, colon, ileum (ND n = 5; HFD n = 6), Epi-AT (ND n = 6; HFD n = 7), and spleen (n = 10 per group). c MAIT cell absolute numbers in the ileum (ND n = 8; HFD n = 10) and MAIT cell number per gram of Epi-AT (ND n = 13; HFD n = 12). d MAIT cell frequency kinetic analysis of mice fed ND or HFD in the ileum (0 weeks n = 8; 6 weeks ND n = 11, HFD n = 14; 12 weeks ND n = 9, HFD n = 10; 16 weeks ND n = 10, HFD n = 13) and Epi-AT (0 weeks n = 8; 6 weeks ND n = 11, HFD n = 14; 12 weeks ND n = 15, HFD n = 15; 16 weeks ND n = 17, HFD n = 19). Pooled data from four independent experiments are represented. e Migration of MAIT cells into lean or obese recipient mice after 12 weeks of ND or HFD. Purified MAIT cells from CD45.1 Va19 transgenic Ca-/- mice were transferred into CD45.2 mice and analyzed 5 days later. Graphs represent CD45.1 MAIT cells frequency in the ileum (n = 10 per group) and Epi-AT (n = 10 per group). Pooled data from two independent experiments are represented. f Ki67 staining of MAIT cells from the ileum (ND n = 8; HFD n = 10) and Epi-AT (ND n = 10; HFD n = 10) of 12 weeks HFD or ND-fed mice. Pooled data from two independent experiments are represented. g qPCR analysis of apoptotic (cMyc, Caspase9, and Bax) and anti-apoptotic (Bcl-2) gene expression by FACS-sorted MAIT cells from mice fed ND (n = 8) or HFD (n = 9) for 12 weeks. Pooled data from two independent experiments are represented. h BCL-2 MFI in MAIT cells from the ileum and Epi-AT from mice fed ND or HFD for 12 weeks (n = 4 per group). For b, c, e, and f, each symbol represents an individual mouse (small horizontal lines indicate the mean ± S.E.M.). In d, g, and h, data are represented as mean ± S.E.M. Statistical analysis were performed by two-tailed Mann–Whitney test, *P < 0.05, **P < 0.01, ***P < 0.001 (see also Supplementary Fig. 1).

Fig 5: PTP1B deletion enhances T-cell activation, proliferation, survival, and STAT5 signaling. A, Naïve (CD44loCD62Lhi) CD8+ T cells from Ptpn1fl/fl and Lck-Cre;Ptpn1fl/fl were stimulated with plate-bound a-CD3 (2.5 µg/mL) and a-CD28 (1.25 µg/mL) for 48 hours and CD25, CD44, CD69, and CD62L levels determined by flow cytometry. B, CTV-labeled naïve CD8+ T cells from Ptpn1fl/fl and Lck-Cre;Ptpn1fl/fl mice were stimulated with the indicated concentrations of plate-bound a-CD3 plus a-CD28 (1.25 µg/mL) for 72 hours and T-cell proliferation (CTV dilution) determined by flow cytometry. C, Basal intracellular p-STAT5 and BCL2 levels were assessed by flow cytometry in naïve (CD44lo) and memory (CD44hi) CD4+ and CD8+ T cells from Ptpn1fl/fl and Lck-Cre;Ptpn1fl/fl mice. D, Intracellular Eomes and T-bet protein levels were assessed by flow cytometry in CD8+ and CD4+ T cells from Ptpn1fl/fl and Lck-Cre;Ptpn1fl/fl mice. E,In vitro–generated rested effector CD8+ or CD4+ T cells were stimulated with IL2 (5 ng/mL) for the indicated times and intracellular STAT5 Y694 phosphorylation (p-STAT5) assessed by flow cytometry. Representative results (means ± SEM) from at least two independent experiments are shown. Significance in A–D was determined using a two-tailed Mann–Whitney U test and in E using a two-way ANOVA test. *, P < 0.05; **, P < 0.01; ***, P < 0.001.