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. 2022 Feb 17;119(8):e2111064119. doi: 10.1073/pnas.2111064119

Fig. 1.

Fig. 1.

Generation of TFH from CD4 effectors requires Ag recognition during the effector checkpoint. (A) Experimental design: Naive OT-II.Thy1.1+ cells were transferred into PR8-OVAII-infected mice (first hosts). At 6 dpi, OT-II.Thy1.1+ effectors were isolated from first hosts and transferred into the following groups of second hosts: 6-dpi PR8-OVAII-infected, 6-dpi PR8-infected, or uninfected mice. Donor cells were analyzed 8 dpi. FACS, fluorescence-activated cell sorting. (B) Percentage and numbers of spleen donor TFH (CXCR5+Bcl6+). (C) Percentage and number of spleen donor GC-TFH (GL7+CXCR5+Bcl6+). (D and E) Representative histogram of ICOS (D) and PD1 (E) expression by spleen donor cells (negative control: naive CD4 from uninfected mice). ICOS normalized median fluorescence intensity (nMFI) (D) and PD1 nMFI (E) expression by spleen donor cells. (A–E, n = 10 per group pooled, 2 independent experiments). Error bars represent SEM. Statistical significance was determined by two-tailed, unpaired Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.001).