Figure 4.

Immune cell subsets of CCR7^−/− mice exhibit an activated and inflammatory phenotype within the GM and mLNs. A: Arithmetic means of absolute gastric CD19⁺ B cells and CD4⁺ and CD8⁺ T cells in the GM tissue of Wt (black bars; n = 12 to 13) and CCR7^−/− (open bars; n = 5) mice. Data were calculated from flow cytometric analysis of CD3⁺CD4⁺ and CD3⁺CD8⁺ expression for T-cell subtypes and CD19⁺ expression for B cells of size-gated gastric lymphocytes. B: Flow cytometric analysis of activation and maturation markers on CD19⁺-gated B cells isolated from the GM of Wt (black curve; n = 6 to 9) or CCR7^−/− mice (n = 7 to 10; red curve: isotype control). GM-derived (C) or mLN-derived (D) T-cell subsets (gated on CD4⁺ or CD8⁺) of 6- to 10-month-old CCR7^−/− (GM, n = 12; mLN, n = 12 to 14) and Wt mice (GM, n = 9; mLN, n = 6 to 8) were stained for CD69 expression and analyzed by flow cytometry (shaded curve: isotype control). Total numbers of activated CD69⁺ CD44^high T-cell subsets in GM (C; right panel) and percentages of activated CD69⁺ CD44^high T-cell subsets in mLN (D; right panel) of Wt and CCR7^−/− mice are shown. E: Flow cytometric analysis of IFN-γ or IL-17A production in GM-derived CD4⁺ T cells from Wt (n = 9) and CCR7^−/− mice (n = 3) after in vitro culture with (+) or without (-) brefeldin, ionomycin, and PMA. Total numbers of gastric CD4⁺ IFN-γ⁺ or CD4⁺ IL-17A⁺ cells are shown. F: Serum samples from 4- to 6-week-old Wt (black bar; n = 9) and CCR7^−/− mice (open bar; n = 9) were analyzed for the T_H17 cytokine IL-17A by enzyme-linked immunosorbent. Bars in A–E represent mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001; Mann-Whitney U-test.