Figure 7.

Lymphocyte subset analysis of spleens from CD6^−/− and CD6^+/+ mice undergoing chronic graft-versus-host disease (cGvHD)-induced lupus-like disease. (A) Flow cytometry contour plots of B220⁺ cells and percentage (mean ± SD) of B220⁺, CD4⁺, and CD8⁺ cells from total spleens harvested at week 5 post cGvHD induction in CD6^−/− (n = 6; solid bars) and CD6^+/+ (n = 6; empty bars) mice. (B) Flow cytometry contour plots and percentages (mean ± SD) of spleen B220⁺ cells with marginal zone B cells (MZ, CD21⁺CD23^low), follicular (B2, CD21⁺CD23⁺), and transitional type 1 (T1, CD21⁻CD23⁻) phenotypes from the same groups of mice as in (A). (C) Percentages (mean ± SD) of memory (CD19⁺CD138⁻IgD⁻IgM⁻), plasma (CD19⁻ CD138⁺), and germinal zone B-cells (CD19⁺GL-7⁺) from the same groups of mice as in (A). (D) Percentages (mean ± SD) of follicular helper (T_FH; CD4⁻CXCR5⁺PD1⁺) and follicular regulatory T cells (T_reg) (T_FR; CD4⁻CXCR5⁺PD1⁺ FoxP3⁺) from the same groups of mice as in (A). (E) Left, Percentage (mean ± SD) of CD4⁺ spleen T cells with regulatory (CD25⁺FoxP3⁺) and activated (CD25⁺FoxP3⁻) phenotypes from the same groups of mice as in (A). (E) Right, Increment in the percentage of spleen T_reg (CD4⁺CD25⁺FoxP3⁺) cells at week 5 postinduction of cGvHD in the same groups of mice as in (A). Results are expressed as the ratio (mean ± SD) resulting from dividing the percentage of T_reg at week 5 by that at week 0. (F) Flow cytometry contour plots and percentage (mean ± SD) of BrdU⁺ cells from total, B220⁺ and CD4⁺ splenocytes at week 5 from the same groups of mice as in (A). (G) Percentage (mean ± SD) of total spleen cells with late-stage apoptotic phenotype (Annexin V⁺ 7-AAD⁺) from the same groups of mice as in (A). In all cases, data shown are from one representative experiment of three performed. n.s., not significant; BrdU, bromodesoxyuridine; **p < 0.01; ***p < 0.001 (Student’s t-test).