Extended Data Fig. 5. Migrated B cells rapidly activate in the vagina and stay for several weeks.

a, Naive C57BL/6 mice or C57BL/6 mice immunized with TK⁻ HSV-2 5 wk prior were challenged with WT HSV-2. Eight, sixteen and twenty-four hours after challenge, CD86 and CD69 expression on IgD⁻IgG⁺ memory B cells in vaginal tissues was analyzed by flow cytometry. b, C57BL/6 mice were immunized vaginally with TK⁻ HSV-2 five weeks prior. After challenge, frozen sections of vagina were stained with antibodies against IgG (red), CD4, HSV, CD31 and VCAM-1 (green), and DAPI (blue). Scale bars indicate 100 μm. c, C57BL/6 mice immunized with TK⁻ HSV-2 5 wk prior were challenged with WT HSV-2. At the indicated days after challenge, the number of memory B cells in vaginal tissues was analyzed by flow cytometry (n=2–4 mice in each time point). Data are mean ± SEM. Data are representative of two independent experiments.