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. Author manuscript; available in PMC: 2015 Jan 12.
Published in final edited form as: J Immunol. 2014 Jun 13;193(2):746–756. doi: 10.4049/jimmunol.1302848

FIGURE 4.

FIGURE 4

B cell depletion impairs T cell expansion, memory conversion, and virus clearance during LCMV infection. Naive 4-mo-old mice were given control (closed bars) or CD20 mAb (open bars) 7 d prior to treatment with PBS or infection with LCMV. Splenocytes harvested 7 d later were assessed by immunofluorescence staining with flow cytometry analysis of viable, single lymphocytes. Representative panels show B220 versus CD4 staining (left panels) and B cell, CD4+, and CD8+ T cell numbers (right panels) (A) and CD25 versus intracellular Foxp3 staining for CD4+ T cells (left panels) and Treg numbers (right panels) (B). (C) B cell depletion reduces LCMV viral clearance. Spleens from control and CD20 mAb-treated mice infected with LCMV were harvested 7 d postinfection, with viral RNA levels quantified by RT-PCR. Bar graph values represent mean (± SEM) relative LCMV RNA expression levels (n = 4 mice/group in three independent experiments). (D–G) B cell depletion impairs LCMV-driven T cell activation and effector subset conversion. Representative panels show CD44 versus CD62L staining (left panels) for spleen CD4+ (D) and CD8+ T cells (F), with cell numbers for the spleen and lymph nodes shown (right panels). (E) Representative panels show PSGL1 versus Ly6C staining (left panels) for spleen and lymph node CD4+ cells, with activated CD4+ T cell conversion to effector (Eff; Ly6C+PSGL1+), prememory (Pre-M; Ly6C− PSGL1+), and T follicular helper (Tfh; Ly6C− PSGL1−) numbers shown (right panels). (G) Representative dot plots show CD127 versus KLRG1 staining (left panels) for spleen and lymph node CD8+ T cells, with KLRG1+ effector cell numbers shown (right panels). (A, B, D, and F) Graphs show mean (± SEM) numbers of the indicated cell types from mice treated with control or CD20 mAb (n = 21–26 mice/group in four to five independent experiments). (E and G) Graphs show results from individual mice (n = 5 mice/group), with means indicated by horizontal bars. (A–G) Mean (± SEM) cell frequencies within the indicated dot plot gates are shown. Significant differences between sample means are indicated: *p < 0.05, **p < 0.01, ***p < 0.001. Ctrl, control.