Fig. 2

Location and activation status of GL7⁺ and GL7⁻ antigen-specific B cells in Peyer’s patch (PP) germinal centers (GC). a Representative microscopy image showing that NP-specific GFP⁺ B cells (green) localize both to the light, CD86⁺ (red), and dark, CXCR4⁺ (blue), zones of the GC (GL7⁺) in PPs following p.o. immunization with NP-CT. b Flow cytometry dot-plots demonstrating expression of light (CD83⁺) and dark (CXCR4⁺) zone phenotypic markers in GL7⁺ and GL7⁻ GFP⁺ and endogenous GL7⁺ GFP⁻ B cells using the gating scheme in Supplementary Fig. 2a. c, d Flow cytometry histograms and heat maps depicting expression of surface markers on GL7⁺ or GL7⁻ NP-specific GFP⁺ or endogenous GFP⁻ B cells in PP following a single p.o. immunization. c Representative flow data demonstrating the expression of CD62L, CD95 and CD23. d Heatmaps depicting the relative expression (MFI[sample]/MFI[maximum for any sample]) for the indicated markers in three mice. e Model used for adoptive transfer of sorted GL7⁺ or GL7⁻ activated GFP⁺ B cells from PP to a “naïve” host that had been given NP-CT 24 h prior to cell transfer. f, g At 8–10 days following the transfer of sorted activated GFP⁺ B cells, the frequency of activated GFP⁺ B cells of all CD19⁺ B cells in PPs of the recipient mice and their distribution in GL7⁺ and GL7⁻ cell populations were determined using flow cytometry. h A representative image demonstrating the GC location of sorted and transferred GL7⁺ or GL7⁻ GFP⁺ (green) B cells in the PP of recipient mice. Sections were stained with B220 (red) and GL7 (white) antibodies to reveal the position of the GC. These are representative data from at least three independent experiments with 3–5 mice in each experiment giving similar results. Source data are provided as a Source Data file