Fig. 1: Functionally heterogeneous T_FH phenotypes are induced by diverse pathogen infections.

a–i, Analysis of draining lymph node cells from wild-type (a–c,h,i) and ZsGreen_T-bet reporter (d–g) mice infected with the indicated pathogens at the early peak GC response (LCMV day 12, influenza day 10, T. muris day 21, H. polygyrus day 12 and C. rodentium day 12). Representative plots of CD4⁺CD44⁺Ly6C⁺CD162⁺ T_eff cells and CD4⁺CD44⁺CXCR5⁺PD-1⁺Ly6C⁻CD162⁻ T_FH cells with histograms displaying Bcl-6 expression (a). Frequencies of T_FH cells in CD4⁺CD44⁺ gate (b) and the ratio of T_FH:T_eff cells (LCMV n = 8, influenza n = 8, T. muris n = 6, H. polygyrus n = 9, C. rodentium n = 10 mice per group) (c). Representative plots of ZsGreen_T-bet reporter expression (T_FH cells are blue, T_eff cells are red) (d). ZsGreen_T-bet reporter⁺ frequency and gMFI of CD4⁺CD44⁺CXCR5⁺PD-1⁺Ly6C⁻CD162⁻ T_FH cells (LCMV n = 8, influenza n = 8, T. muris n = 6, H. polygyrus n = 10, C. rodentium n = 7 mice per group) (e). Immunofluorescence staining of draining lymph node GCs. Red arrows indicate ZsGreen_T-bet reporter⁺ T_FH cells. Yellow, CD4; blue, IgD; magenta, GL7; green, ZsGreen_T-bet reporter. Scale bar, 200 μm (f). Correlation of ZsGreen_T-bet reporter expression with the T_eff:T_FH cell ratio across infections (g). Frequency of T_FH cells that produced IFNγ, IL-4 or IL-17A (h). IFNγ⁺ T_FH1, IL-4⁺ T_FH2, and IL-17⁺ T_FH17 cells are included in the entire T_FH population (i). The inner slice displays cytokine coexpression. The outer ring (green) indicates the proportion of ZsGreen_T-bet⁺ T_FH cells expressed. The data are from 6–10 mice per group and are presented as the mean ± s.e.m. Statistical tests included one-way analysis of variance (ANOVA) for multiple comparisons and Pearson correlation with two-tailed P values. ****P < 0.0001 or otherwise indicated.

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