1. Identification of CD25− Tfr cells.
Identification of CD25− Tfr cells. Mice were vaccinated s.c. with 100 μg of NP-Ova (Biosearch) in alum, and draining LNs (dLNs) or Peyer’s patches were taken at day 7 or the indicated time. (A) Gating strategy of CD25+ and CD25− Tfr cells. Cells were first gated as CD3+CD4+B220−CD11c−CD11b−Live/Dead-dye− before the start of the shown gating. The fluorescence minus one (FMO) control is CD44+CD62L− Treg cells with anti-CXCR5 omitted. Data are representative of >10 separate experiments. Max, maximum. (B) Proportion of CD25− cells in gated CXCR5 and PD-1 low to high populations in Peyer’s patch CD44+CD62L− Treg cells. Data are representative of >10 separate experiments. (C) Time course of GC, Tfh, CD25+ Tfr, and CD25− Tfr proportions in dLNs following vaccination at day 0. (D) Tfr cells in BCL6flox/flox and BCL6wt/wt CD4-Cre mice, with Peyer’s patch T cells pregated on CD44+CD62L− Treg cells. (C and D) Data are pooled from three mice, representative of two separate experiments.
James Badger Wing et al. PNAS doi: /pnas
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