Fig 5. CNS–28 is critical for type 1 responses during host defense and inflammation.

(A) Body weight of Rag2^−/− mice transferred i.p. with CD4⁺CD25⁻CD62L⁺ cells from WT or CNS–28^Δ mice.

(B) Hematoxylin and eosin staining of colonic tissue from the different groups as in (A) 10 weeks after colitis induction, scale bar, 50μm.

(C) Left: Quantification of pathological changes in the colon of mice as in (A); Right: Colon lengths of Rag2^−/− mice which had received the indicated cells for transfer as in (A), measured from the colocecal junction to the anal verge

(D-E) (D)Flow cytometry analysis and (E) Quantification of IFN-γ expression by CD4⁺ T cells isolated from LP from indicated groups as in (A) 10 weeks after colitis induction.

(F-I) Splenocytes from WT and CNS–28^Δ mice were isolated and cultured in the presence of IL-12 and IL-2 for 6 h. IFN-γ expression in (F, G) CD8⁺ T cells or (H, I) NK1.1⁺ NK cells were measured by (F, H) flow cytometry or (G, I) Quantification.

(J-M) Listeria were inoculated into WT and CNS–28^Δ mice by oral gavage. The mice were sacrificed on day 14 for further tests.

(J) IFN-γ level in serum was assessed by ELISA at indicated day.

(K) Bacteria CFU was counted at day 7 after infection in liver and spleen.

(L) Flow cytometry analysis and (M) Quantification of IFN-γ expression in CD4⁺, CD8⁺ T cells from spleen 7 days after infection and NK cells isolated from spleen 1 day after infection.

Data are representative of at least two independent experiments (A, B, C-M) or pooled from two independent experiments (C). *p < 0.05, ^**p < 0.01 (Student’s t test, error bars represent SD).