Extended Data Fig. 4: Tfr cell reduction in aged mice does not enhance GC responses.

(a) Representative flow cytometry plots identifying Tfr cells (PD1⁺CXCR5⁺CD4⁺Foxp3⁺B220⁻) in the iLNs of adult and aged C57BL/6 mice 10 days post-immunization with 1W1K-NP in alum. The frequency (b) and total number (c) of Tfr cells in adult and aged C57BL/6 mice was quantified at days 0, 7,10, 14 and 21 after immunization with 1W1K-NP in alum. The data is representative of two independent experiments (n = 14) where each symbol represents the mean ± SD and p-values were generated by performing a two-way ANOVA with Sidak’s multiple comparisons test. (d) Representative flow cytometry plots indicating deletion of Tfr cells in aged 90–97-week-old Foxp3^creCxcr5^fl/fl mice compared to aged littermate control Foxp3^cre mice 10 days after immunization with NP-KLH in alum. Quantification of the frequency (e) and total number (f) of Tfr cells (n = 12). (g) Representative flow cytometry plots identifying GC B cells (GL7⁺CD38⁻B220⁺GR-1^-) in Foxp3^creCxcr5^fl/fl mice 10 days after immunization with NP-KLH in alum. Quantification of the frequency (h) and total number (i) of GC B cells (n = 12). The values next to the gates on flow cytometry plots indicate the population percentage. The data is representative of two independent experiments where each symbol represents a mouse, and the bar height represents the median. The p-values were obtained by performing an unpaired, two-tailed Mann-Whitney U test.

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