Figure 4. T Cell-Intrinsic IL-27 Signaling Is Essential for the Development of T1D.

(A) T1D incidence of NOD and NOD.Il27ra^−/− mice. ***p < 0.005 by log rank test.

(B) Summary of insulitis in female NOD and NOD.Il27ra^−/− mice.

(C) Summary of insulitis in male NOD and NOD.Il27ra^−/− mice. Pancreatic islets were scored for insulitis as in Figure 1. Each symbol represents one mouse. The horizontal bar depicts the mean. **p < 0.01 by Mann-Whitney test. NS, not significant.

(D) T1D incidence study of sublethally irradiated NOD.Il27^−/− or NOD.Il27ra^−/− females infused with BM cells (5 × 10⁶) from sex-matched NOD.Rag1^−/− mice as indicated. **p < 0.01 by log rank test.

(E–G) Incidence of T1D in recipients of adoptively transferred T cells.

(E) Splenic T cells (5 × 10⁶) were isolated from 6-week-old NOD females and transferred into sex-matched NOD.Rag1^−/− or NOD.Rag1^−/−.Il27ra^−/− recipients. T1D incidence was not significantly different between recipient groups.

(F) Splenic T cells (5 × 10⁶) were isolated from 6- to 11-week-old NOD or 11- to 15-week-old NOD.Il27ra^−/− females and transferred into sex-matched NOD.Rag1^−/− recipients. ***p < 0.005 by log rank test.

(G) Splenic CD4 (4 × 10⁶) and CD8 (2 × 10⁶) T cells were isolated from indicated 6- to 8-week-old female strains and co-transferred into sex-matched NOD.Rag1^−/− recipients. **p < 0.01 and ***p < 0.005 by log rank test.

(H) In vitro suppression function of NOD and NOD.Il27ra^−/− Tregs. Splenic CD4⁺CD25⁻ T cells were isolated from NOD mice, labeled with CFSE, and cultured either alone or in the presence of unlabeled CD4⁺CD25⁺ Tregs at the indicated ratios. Cells were activated with soluble anti-CD3 in the presence of NOD.Rag1^−/− splenocytes for 3 days, and proliferation was analyzed using flow cytometry. Representative histograms of CFSE dilution from one experiment are shown in Figure S4. Summarized data from three independent experiments are shown. The percentage suppression was calculated as [percentage of divided CD4⁺CD25⁻ T cells (without Tregs) – percentage of divided CD4⁺CD25⁻ (with Tregs)]/[percentage of divided CD4⁺CD25⁻ T cells (without Tregs)] 3 100. Error bars represent SEM. The suppression function of NOD and NOD.Il27ra^−/− Tregs was not significantly different.

(I) Incidence of T1D in recipients of adoptively transferred T cells. Splenic CD25⁻ T cells (5 × 10⁶) isolated from 13- to 15-week-old NOD females and splenic CD4⁺GITR⁺CD25⁺ Tregs (5 × 10⁵) FACS sorted from 6- to 9-week-old NOD or NOD.Il27ra^−/− females were co-transferred into sex-matched NOD.Rag1^−/− recipients. ***p < 0.005 by log rank test. NS, Not significant.

See also Figures S1, S3, and S4.