Extended Data Fig. 7: Pathogen-induced cytokine signaling influence T_FH and T_EFF bifurcation, T_FH subpopulations, and T_FH cells capacity to direct B cell and antibody output.

a, Gating strategy to identify CD4⁺CD44⁺CXCR5⁺PD-1⁺Ly6C⁻CD162⁻ T_FH1, T_FH2, T_FH17, and T_FH17.1 phenotypes based on expression of CXCR3 and CCR6 chemokine receptors. b-k, Analysis of draining lymph nodes from (b,c) wild-type, (d) Tgfbr2-Lck^Cre and Lck^Cre mice, (f) Ifnar^−/− and Ifnar^+/+ mice, (e,h,j,k) wild-type and Tgfbr2-Lck^Cre 50:50 bone marrow chimeras, or (g-i,k) wild-type and Ifnar^−/− 50:50 bone marrow chimera mice infected with (b,d,e,h,i,k) influenza or (c,f,g,h,j,k) LCMV. (b,c) gMFI expression of IFNg, IL-4, and IL-17 in T_FH1 (CXCR3⁺CCR6⁻), T_FH2 (CXCR3⁻CCR6⁻), T_FH17 (CXCR3⁻CCR6⁺) and T_FH17.1 (CXCR3⁺CCR6⁺) populations (influenza n = 10, LCMV n = 6 mice per group). (d,f) T_FH and T_EFF cell counts, T_FH and T_EFF cell frequencies, and T_FH/T_EFF cell ratio (influenza n = 9-10, LCMV n = 7 mice per group). (e,g) T_FH and T_EFF cell frequencies and T_FH/T_EFF cell ratio (influenza n = 10, LCMV n = 8 mice per group). (h) Comparison of effect of Tgfbr2-Lck^Cre and Ifnar ^−/− on T_FH frequency in chimeras. (i-j) Frequency of T_FH1, T_FH2, T_FH17 and T_FH17.1 cells within T_FH population in chimeric mice (influenza n = 8, LCMV n = 7 mice per group). (k) Comparison of effect of Tgfbr2-Lck^Cre and Ifnar ^−/− on T_FH1, T_FH2 and T_FH17 cell frequency in influenza and LCMV infection in chimeras (Tgfbr2-Lck^Cre influenza n = 10, Tgfbr2-Lck^Cre LCMV n = 7, Ifnar ^−/− influenza n = 8, Ifnar ^−/− LCMV n = 8 mice per group). l, Gating strategy to identify IgG1 and IgG2 class switched GC B cells, and dark zone (DZ) and light zone (LZ) GC B cells. Data show experiments of 6–10 mice per group and mean ± SEM. Statistical tests: Two-tailed unpaired Student’s t test for intact system; two-tailed paired Student’s t test for bone marrow chimera model. **** P values are <0.0001 or otherwise indicated.

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