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Human B cells promote T-cell plasticity to optimize antibody response by inducing coexpression of TH1/TFH signatures  Jelle de Wit, PhD, Tineke Jorritsma,

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Presentation on theme: "Human B cells promote T-cell plasticity to optimize antibody response by inducing coexpression of TH1/TFH signatures  Jelle de Wit, PhD, Tineke Jorritsma,"— Presentation transcript:

1 Human B cells promote T-cell plasticity to optimize antibody response by inducing coexpression of TH1/TFH signatures  Jelle de Wit, PhD, Tineke Jorritsma, BSc, Mateusz Makuch, MSc, Ester B.M. Remmerswaal, BSc, Hanny Klaasse Bos, BSc, Yuri Souwer, PhD, Jacques Neefjes, PhD, Ineke J.M. ten Berge, MD, PhD, S. Marieke van Ham, PhD  Journal of Allergy and Clinical Immunology  Volume 135, Issue 4, Pages (April 2015) DOI: /j.jaci Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 T cells are polarized toward an IFN-γ+/IL-21+ phenotype. B cells were incubated with anti-IgM coated Salmonella species (Sal; A and B) or 0.1 μg/mL TT (C and D) and cocultured with autologous CD4+ T cells. T-cell polarization was measured at day 11 (n = 6). *P < .05, **P < .01, and ***P < .005. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 B cells, but not DCs, induce IL-21 in T cells. CD4+ T-cell polarization induced by autologous anti-IgM Salmonella-primed B cells (left) or anti-IgM Salmonella species–primed DCs (right) was measured at day 11 in 1 representative experiment (A) and combined in 11 different donors (B): B cells (open and solid bars) versus DCs (hatched bars). *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Blocking of IL-21 reduces both IgM and IgG antibody secretion by B cells. A, B cells with anti-IgM Salmonella were cultured in the presence of IFN-γ, IL-4, or IL-21. Proliferation was measured at day 6 (n = 4). B, Antibody secretion was measured at day 12 (n = 4). C, Antibody secretion by anti-IgM Salmonella-primed B cells cultured with anti–IL-21, anti–IFN-γ, or anti–IL-4 blocking antibodies at day 12 (n = 4). *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 B cells induce IL-21 in both naive and memory CD4+ T cells. B cells with anti-IgM Salmonella species were cocultured with naive or memory CD4+ T cells. A, Proliferation of T cells at day 6 (n = 7). B and C, T-cell polarization at day 11 (n = 11). D, CD4+ T cells activated by Salmonella-primed B cells were sorted at 11 days and restimulated for 11 days by B cells with Salmonella (Bsal) or DCs with Salmonella (DCsal) or through anti-CD3 and anti-CD28 antibodies (3/28). Representative plots are shown for n = 4 (see Fig E6). *P < .05, **P < .01, and ***P < .005. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 T cells isolated from human LNs show IFN-γ+/IL-21+ expression. A and B, Ex vivo cytokine production of mononuclear cells from PBMCs and isolated LNs from the same donors. C, Percentages of IFN-γ+ SP, IFN-γ+/IL-21+ DP, and IL-21+ SP populations in CD4+ T cells in PBMCs and LNs (n = 5). Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 IFN-γ+/IL-21+ DP CD4+ T cells express both T-bet and Bcl-6. B cells with anti-IgM Salmonella were cultured with naive CD4+ T cells and analyzed on day 11 for expression of Bcl-6, T-bet (A; n = 7), CXCR5, ICOS, and PD-1 (B; n = 4). IL-21+ SP (red), IFN-γ+/IL-21+ DP (blue), and IFN-γ+ SP (orange) cells. Bars show the difference in median fluorescent intensity (MFI) between subsets and negative (IFN-γ−/IL-21−) conditions. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 7 Blocking of IL-6 reduces IL-21+ TFH cell differentiation of naive T cells. A, B cells with and without anti-IgM Salmonella species were cocultured with T cells. After 24 hours, B cells were separated, and IL-12p40 and IL-6 mRNA levels were analyzed (n = 8). B and C, Naive T-cell differentiation by Salmonella-positive B cells was measured after 11 days of coculture with anti–IL-12p35 mAb, anti–IFN-γ (Fig 7, B; n = 5), anti–IL-6, and/or anti–IL-21 (Fig 7, C; n = 7) blocking antibodies. *P < .05, **P < .01, and ***P < .005. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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