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Innate B cells are capable of plasma cell differentiation and IgA class-switching in response to TLR4 stimulation. (A) Purified total splenic B cells from <t>Blimp1-YPF</t> reporter mice were stimulated for 3 days with anti-IgM or LPS and analyzed by flow cytometry for the differentiation into Blimp1(YFP) + plasma cells. (B) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) from the spleen of Blimp1-YPF reporter mice were stimulated with LPS and analyzed for the formation of Blimp1(YFP) + CD138 + plasma cells (day 3). (C–E) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) were stimulated as indicated with LPS (day 3), anti-IgM (day3) or anti-IgM + anti-CD40 + IL-4 (day 6) and culture supernatants were analyzed by ELISA for (C) IgM and (D, E) IgA antibody levels. Lines indicate mean ± SEM; nd , not detectable; * p < 0.5; ** p < 0.01; *** p < 0.001; one-way ANOVA and Tukey’s multiple comparisons test. Data are representative of at least 3 independent experiments.
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Innate B cells are capable of plasma cell differentiation and IgA class-switching in response to TLR4 stimulation. (A) Purified total splenic B cells from <t>Blimp1-YPF</t> reporter mice were stimulated for 3 days with anti-IgM or LPS and analyzed by flow cytometry for the differentiation into Blimp1(YFP) + plasma cells. (B) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) from the spleen of Blimp1-YPF reporter mice were stimulated with LPS and analyzed for the formation of Blimp1(YFP) + CD138 + plasma cells (day 3). (C–E) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) were stimulated as indicated with LPS (day 3), anti-IgM (day3) or anti-IgM + anti-CD40 + IL-4 (day 6) and culture supernatants were analyzed by ELISA for (C) IgM and (D, E) IgA antibody levels. Lines indicate mean ± SEM; nd , not detectable; * p < 0.5; ** p < 0.01; *** p < 0.001; one-way ANOVA and Tukey’s multiple comparisons test. Data are representative of at least 3 independent experiments.
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Innate B cells are capable of plasma cell differentiation and IgA class-switching in response to TLR4 stimulation. (A) Purified total splenic B cells from Blimp1-YPF reporter mice were stimulated for 3 days with anti-IgM or LPS and analyzed by flow cytometry for the differentiation into Blimp1(YFP) + plasma cells. (B) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) from the spleen of Blimp1-YPF reporter mice were stimulated with LPS and analyzed for the formation of Blimp1(YFP) + CD138 + plasma cells (day 3). (C–E) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) were stimulated as indicated with LPS (day 3), anti-IgM (day3) or anti-IgM + anti-CD40 + IL-4 (day 6) and culture supernatants were analyzed by ELISA for (C) IgM and (D, E) IgA antibody levels. Lines indicate mean ± SEM; nd , not detectable; * p < 0.5; ** p < 0.01; *** p < 0.001; one-way ANOVA and Tukey’s multiple comparisons test. Data are representative of at least 3 independent experiments.

Journal: Frontiers in Immunology

Article Title: Acquisition of innate B cell properties and generation of autoreactive IgA antibodies by follicular B cells during homeostatic proliferation

doi: 10.3389/fimmu.2025.1506628

Figure Lengend Snippet: Innate B cells are capable of plasma cell differentiation and IgA class-switching in response to TLR4 stimulation. (A) Purified total splenic B cells from Blimp1-YPF reporter mice were stimulated for 3 days with anti-IgM or LPS and analyzed by flow cytometry for the differentiation into Blimp1(YFP) + plasma cells. (B) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) from the spleen of Blimp1-YPF reporter mice were stimulated with LPS and analyzed for the formation of Blimp1(YFP) + CD138 + plasma cells (day 3). (C–E) FACS-sorted B cell subsets (FoB, MZB, and B1a B cells) were stimulated as indicated with LPS (day 3), anti-IgM (day3) or anti-IgM + anti-CD40 + IL-4 (day 6) and culture supernatants were analyzed by ELISA for (C) IgM and (D, E) IgA antibody levels. Lines indicate mean ± SEM; nd , not detectable; * p < 0.5; ** p < 0.01; *** p < 0.001; one-way ANOVA and Tukey’s multiple comparisons test. Data are representative of at least 3 independent experiments.

Article Snippet: Blimp1-YFP reporter mice (B6.Cg-Tg(Prdm1-EYFP 1Mnz /J); RRID: IMSR_JAX:008828), Rag2 -/- mice (B6.Cg- Rag2 tm1.1Cgn /J; RRID: IMSR_JAX:008449) and C57BL/6J (RRID: IMSR_JAX:000664) were originally obtained from The Jackson Laboratory.

Techniques: Cell Differentiation, Purification, Flow Cytometry, Enzyme-linked Immunosorbent Assay