B cell–helper neutrophils stimulate the diversification and production of immunoglobulin in the marginal zone of the spleen

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  • A Corrigendum to this article was published on 21 January 2014

Abstract

Neutrophils use immunoglobulins to clear antigen, but their role in immunoglobulin production is unknown. Here we identified neutrophils around the marginal zone (MZ) of the spleen, a B cell area specialized in T cell–independent immunoglobulin responses to circulating antigen. Neutrophils colonized peri-MZ areas after postnatal mucosal colonization by microbes and enhanced their B cell–helper function after receiving reprogramming signals, including interleukin 10 (IL-10), from splenic sinusoidal endothelial cells. Splenic neutrophils induced immunoglobulin class switching, somatic hypermutation and antibody production by activating MZ B cells through a mechanism that involved the cytokines BAFF, APRIL and IL-21. Neutropenic patients had fewer and hypomutated MZ B cells and a lower abundance of preimmune immunoglobulins to T cell–independent antigens, which indicates that neutrophils generate an innate layer of antimicrobial immunoglobulin defense by interacting with MZ B cells.

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Figure 1: Neutrophils colonize the splenic MZ and function as NBH cells.
Figure 2: NBH cells interact with splenic MZ B cells to induce CSR, SHM and production of immunoglobulins.
Figure 3: NBH cells include NBH1 and NBH2 subsets distinct from NC cells.
Figure 4: NBH cells activate MZ B cells via BAFF, APRIL and IL-21.
Figure 5: NC cells acquire NBH cell–like properties after exposure to SECs activated by microbial signals.
Figure 6: NBH cells regulate MZ B cells and innate IgM, IgG and IgA responses to microbial TI antigens in vivo.
Figure 7: NBH cells colonize the spleen through a mechanism involving microbial signals.

Change history

  • 12 July 2013

    In the version of this article initially published, the second affiliation for Stephanie C. Ganal is missing. This author is also affiliated with the Spemann Graduate School of Biology and Medicine, Freiburg, Germany. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank J. Farrés, J. Yélamos, A. Muntasell and M. López-Botet (Institut Municipal d'Investigació Mèdica-Hospital del Mar) for reagents, samples and discussions; N. Romo, S. Bascones, E. Ramirez and O. Fornas for help with cell sorting; and S. Mojal for help with the statistical analysis. Supported by Ministerio de Ciencia e Innovación (SAF 2008-02725 to A.Ce.), the US National Institutes of Health (R01 AI074378, P01 AI61093, U01 AI95613 and P01 096187 to A.Ce.), the European Commission (EUROPADnet HEALTH-F2-2008-201549 to A.Ce.), the Juan de la Cierva Program (I.P. and G.M.), the Instituto de Salud Carlos III (C.M.B. and M.G. and A.Ch.), the Ministerio de Ciencia e Innovación (C.M.B. and M.G.), Yerkes National Primate Research Center (P51 RR00165 to G.S.) and Fondazione C. Golgi di Brescia, Associazione Immunodeficienze Primitive (A.P.).

Author information

I.P. and M.Co. designed and did research, discussed data and wrote the paper; C.M.B., B.H. and K.C. designed and did research; L.Ca., M.G., L.Co., A.Cho., M.S., W.X., G.M., A.Chi, T.B. and S.C.G. did research and discussed data; D.M.K., W.T., J.B.B., S.S., J.A.L., B.B., J.L., N.J., F.A., C.D.d.H., N.T., A.Ca., M.T., C.F., V.C., C.C., G.A.D., J.M.B., C.-M.F., G.S., C.C.-R., M.Ca., C.Du., L.D.N., V.L., A.P., J.-L.C., A.D., J.I.A., M.J., J.Y., N.M. and J.D. provided blood and tissue samples and discussed data; and A.Ce. designed research, discussed data and wrote the paper.

Correspondence to Andrea Cerutti.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–22 and Tables 1–6 (PDF 2223 kb)

Supplementary Video 1

NBH cells form NET-like structures to interact with splenic MZ B cells. Confocal microscopy and three-dimensional animation of a spleen section stained for elastase (green), IgD (red) and DNA (blue). The movie was generated by acquiring up to 14 x,y planes with 0.3 μm z spacing. Original magnification, x40. One of three experiments yielding similar results. (AVI 17483 kb)

Supplementary Video 2

NBH cells spontaneously form DNA-containing NET-like projections. Confocal microscopy and three-dimensional animation of NBH cells stained for elastase (green) and DNA (blue). The movie was generated by acquiring up to 25 x,y planes with 0.3 μm z spacing. Original magnification, x63; digital magnification, x2. One of three experiments yielding similar results. (AVI 9943 kb)

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