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. 2022 Mar;142(3Part A):726–731.e4. doi: 10.1016/j.jid.2021.06.038

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© 2021 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Human B cells accumulate and proliferate within cutaneous sites of antigen challenge. (a, b) Frozen sections from skin biopsies taken 1, 3, and 7 days after intradermal challenge with (a) candida and (b) VZV antigens were stained for CD3⁺ T cells (red, left panel), CD20⁺ B cells (red, right panel), and proliferating lymphocytes (Ki67; green) seen within PV areas. All sections were counterstained with DAPI (blue). Original magnification: ×20. Bar = 50 μm. Overall, 3–16 skin biopsies were analyzed per time point (two sections stained per biopsy). Representative images are shown. (c, d) Quantification of CD3⁺ T and CD20⁺ B cells within candida and VZV biopsy sections described in a and b. Day 0 biopsies were taken from healthy skin without previous VZV or candida antigen challenge. Cells were counted within three to five most densely populated PV infiltrates, and the mean number of cells was plotted per PV. Error bars represent mean ± SEM. (e, f) Quantification of proliferating Ki67⁺ CD3⁺ T and Ki67⁺ CD20⁺ B cells within candida and VZV biopsy sections described in a and b. Error bars represent mean ± SEM. Data were analyzed with one-way ANOVA with Tukey’s multiple comparison post-test. (g) Representative immunofluorescence staining of proliferating (CD20⁺Ki67⁺, yellow cells, indicated by white arrows) B cells within day 7 skin biopsy sections (candida, top panel; VZV, bottom panel). Original magnification: ×20. Bar = 50 μm. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. PV, perivascular; VZV, varicella-zoster virus.

Human B cells accumulate and proliferate within cutaneous sites of antigen challenge. (a, b) Frozen sections from skin biopsies taken 1, 3, and 7 days after intradermal challenge with (a) candida and (b) VZV antigens were stained for CD3⁺ T cells (red, left panel), CD20⁺ B cells (red, right panel), and proliferating lymphocytes (Ki67; green) seen within PV areas. All sections were counterstained with DAPI (blue). Original magnification: ×20. Bar = 50 μm. Overall, 3–16 skin biopsies were analyzed per time point (two sections stained per biopsy). Representative images are shown. (c, d) Quantification of CD3⁺ T and CD20⁺ B cells within candida and VZV biopsy sections described in a and b. Day 0 biopsies were taken from healthy skin without previous VZV or candida antigen challenge. Cells were counted within three to five most densely populated PV infiltrates, and the mean number of cells was plotted per PV. Error bars represent mean ± SEM. (e, f) Quantification of proliferating Ki67⁺ CD3⁺ T and Ki67⁺ CD20⁺ B cells within candida and VZV biopsy sections described in a and b. Error bars represent mean ± SEM. Data were analyzed with one-way ANOVA with Tukey’s multiple comparison post-test. (g) Representative immunofluorescence staining of proliferating (CD20⁺Ki67⁺, yellow cells, indicated by white arrows) B cells within day 7 skin biopsy sections (candida, top panel; VZV, bottom panel). Original magnification: ×20. Bar = 50 μm. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. PV, perivascular; VZV, varicella-zoster virus.