Fig. 4: Treatment of wild-type (WT) mice with 17β-estradiol (E2) prevented the induction of follicular helper T (TFH) and germinal center (GC) B cells by NP-OVA immunization.

a, b FACS-sorted naïve CD4 T cells were stimulated by plate-bound anti-CD3 and anti-CD28 antibodies under TFH-like cell skewing conditions for 72 h with or without 20 nM E2. Real-time PCR was performed to measure the mRNA levels of (a) ERα and (b) ERβ. c Experimental scheme of NP (4-hydroxy-3-nitrophenyl acetyl hapten)-conjugated ovalbumin (NP-OVA) immunization and E2 treatment. d, e Flow cytometric analysis of (d) CD4⁺CD44⁺CXCR5⁺Bcl-6⁺ TFH cells and (e) B220⁺CD95⁺GL-7⁺ GC B cells from the inguinal lymph nodes (ILNs) of NP-OVA-immunized DMSO control and E2 treated group mice. f Naive CD4 + T cells from the spleen and ILNs of 6- to 8-week-old WT and CD4-ERα knockout female mice were isolated and stimulated by plate-bound anti-CD3 and anti-CD28 antibodies under TFH-like cell skewing conditions for 72 h. The indicated target mRNA expression levels were quantified by real-time PCR, and the values were compared between the indicated groups. The values represent the mean ± SEM; n = 4. *P < 0.05; and ***P < 0.001