Fig. 3: Induction of cross-reactive hemagglutinin-specific B cells by RBD–np.
From: Mosaic nanoparticle display of diverse influenza virus hemagglutinins elicits broad B cell responses
a, Gating strategy for identifying hemagglutinin-specific B cells by flow cytometry. Anti-CD3, anti-CD8, anti-CD14 and anti-F4/80 were combined and used to separate T cells, monocytes and macrophages (MΦ). A disparate pair of hemagglutinins (NC99 and CA09) was used to define the cross-reactivity of hemagglutinin-specific B cells. b–d, Frequencies of IgD− B cells specific to NC99 hemagglutinin (b) or CA09 hemagglutinin (c) or cross-reactive to NC99 and CA09 hemagglutinin (d) among PBMCs of mice immunized with different RBD–np (n = 15, cumulative data of three independent experiments, except for 8-valent admix group (n = 10, cumulative of two independent experiments). Statistical analyses were done with one-way ANOVA with Tukey’s multiple comparisons post-hoc test by using valence = 1 as a comparator. e, Mutation rate in genes encoding immunoglobulin VH of individually sorted NC99 HA+ B cells isolated from immunized mice (n = 3, NC99, admix and mosaic; n = 2, sequential). Total productive genes obtained were 108, 98, 101 and 36 for the NC99, admix, mosaic and sequential RBD–np groups, respectively. Statistical analysis was done with one-way ANOVA with Tukey’s multiple comparisons post-hoc test.
