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. 2022 May 12:eabq3511. doi: 10.1126/sciimmunol.abq3511

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Copyright © 2022 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works

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Fig. 2. — SARS-CoV-2 S-specific B cell responses induced by BA.1 breakthrough infection. (A-B) Frequency of circulating B cells that recognize recombinant WT and BA.1 RBDs among (A) IgG⁺ and (B) IgA⁺ B cells in BA.1 breakthrough infection donors (n=7) and uninfected/vaccinated donors at one month (n=12) or six months (n=11) following primary vaccination (2x mRNA) or one month following booster mRNA vaccination (3x mRNA, n=11), as measured by flow cytometry. Bars indicate median frequencies. Donors with breakthrough infections occurring after primary mRNA vaccination are shown as squares and those infected after booster mRNA vaccination are shown as triangles. (C) Representative fluorescence-activated cell sorting (FACS) gating strategy used to identify RBD-directed B cells that are WT-specific or WT/BA.1 cross-reactive, shown for a pre-pandemic donor and a breakthrough infection donor. Percentages of WT-specific and WT/BA.1 cross-reactive B cells out of total RBD-reactive cells are shown in parentheses. (D-E) Mean proportions of RBD-reactive B cells that bind WT and/or BA.1 RBDs among (D) total S⁺swIg⁺ B cells or (E) S⁺swIg⁺ CD7⁺ B cells. Error bars represent standard errors of the mean. A median of 65 RBD-specific B cells (ranging 12 to 310 cells) were collected from each donor for this analysis. Samples collected six months following mRNA vaccination were excluded from this analysis due to low numbers of RBD-specific CD71⁺ cells at this time point. (F-H) Percentage of S-reactive swIg⁺ B cells that target the (F) NTD, (G) RBD, and (H) Hexapro-stabilized S2 subunits. Black bars represent median percentages. For breakthrough infection donors, this analysis was restricted to S⁺swIg⁺CD71⁺ B cells to capture the activated response. 87 to 1721 S-reactive B cells were collected from each donor for this analysis. Statistical comparisons were determined by (A-B) two-way ANOVA with subsequent Dunnett's multiple comparisons test or (D-H) two-sided Kruskal-Wallis test with Dunn's multiple comparisons. 1M, one month; 6M, six months; swIg, class-switched (IgG⁺ or IgA⁺) immunoglobulin; WT, wild type. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.