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. 2015 Apr 7;45(6):1706–1715. doi: 10.1002/eji.201445421

Figure 4.

Figure 4

ICOS is required for expansion of antigen‐specific Tfh cells post secondary challenge. (A and B) WT mice were infected with Lm‐2W1S then challenged again at 28 dpi and treated with anti‐ICOS or isotype control mAb 1 day pre‐ and post secondary challenge with Lm‐2W1S. (A) Three days post secondary challenge CD44hi 2W1S:I‐Ab+ CD4+ T cells were stained for CXCR5 and PD‐1 expression and analyzed by flow cytometry. (B) CD44hi 2W1S:I‐Ab+ CD4+ CXCR5− (Teff), CXCR5+ (Tcm precursors), and CXCR5+PD‐1+ (Tfh) cells were enumerated 3 days post secondary challenge. (C and D) Mice were injected with SM1 memory cells and challenged 28 days posttransfer. Enumeration of SM1 memory cells that are CXCR5− (Teff), CXCR5+ (Tcm precursors), and CXCR5+PD‐1+ (Tfh) cells from (C) ICOSL−/− and (D) CD80−/−CD86−/− mice day 4 post secondary challenge. (A) Plots are representative of ≥6 mice from two independent experiments. Values on plots are percentages. (B, C) Graphs show pooled data or (D) representative data from two independent experiments. Each data point represents one mouse; bars show medians. Mann–Whitney test: ns p > 0.05, *p < 0.05, **p ≤ 0.01, ***p ≤ 0.001.