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. 2019 Mar 29;10:606. doi: 10.3389/fimmu.2019.00606

Figure 1.

Figure 1

Transcription factor T-bet is selectively expressed in Type I but not Type II TFH cells. (A–F) WT C57BL/6 mice were infected with LCMV, LM, or immunized with NP-KLH. Lymphocytes of Spleen (for LCMV and LM infection) or draining lymph nodes (for NP-KLH immunization in alum) were isolated and analyzed for T-bet expression in TFH cells at day 8 post infection/immunization. (A–C) Representative flow cytometry of TFH cells (CD44+CXCR5+), Non-TFH cells (CD44+CXCR5−) and Naïve CD4+ T cells (CD44−CXCR5−) in LCMV (A), LM (B), infection or NP-KLH (C) immunization model. Numbers adjacent to outlined areas indicate percent of each subset in parent subset. (D–F) Representative Flow cytometry of T-bet expression in TFH cells, Non-TFH cells and Naïve CD4+ T cells (left) and the summary of T-bet expression by calculating the mean fluorescence intensity (MFI) of T-bet in each cell subsets (right) during LCMV (D), LM (E) infection, or NP-KLH (F) immunization in alum. (G,H) WT SMARTA cells (CD45.1+) were transferred into WT naïve C57BL/6 mice (CD45.2+) and the splenocytes were analyzed for T-bet expression of virus-specific TFH and TH1 cells at day3, 6, 10, 40, 90, and 160 post LCMV infection. (G) Representative Flow cytometry of T-bet expression in TFH cells (CD45.1+CXCR5+), TH1 cells (CD45.1+CXCR5−) and Naïve CD4+ T cells (CD45.2+CD44−). (H) Kinetics of T-bet expression of TFH and TH1 cells by calculating the MFI of T-bet in each subset followed by normalization to the T-bet MFI of Naïve CD4+ T cells. ns, not significant; *P < 0.05, **P < 0.01,****P < 0.0001 (unpaired two-tailed t-test). Data are representative of two independent experiments with 3–5 mice per group (error bars, SEM).