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Regulatory B Cells in Allergic Airways Disease and Asthma

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Regulatory B Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1190))

Abstract

B cells have long been known to participate in both innate and adaptive immune responses by contributing to antigen presentation and by producing antigen-specific antibodies. Recent evidence shows that certain B-cell subsets can also inhibit T-cell immune responses. Like regulatory T cells (Treg), these regulatory B cells (Breg) appear to comprise several subpopulations. How Breg cells are generated and how they control immune responses in vivo are just beginning to be elucidated. Here, we provide detailed instructions for the identification, isolation, and functional characterization of Breg cells in a murine model of allergic airway disease.

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References

  1. Liu Y, Wu Y, Ramarathinam L, Guo Y, Huszar D, Trounstine M, Zhao M (1995) Gene-targeted B-deficient mice reveal a critical role for B cells in the CD4 T cell response. Int Immunol 7:1353–1362

    Article  CAS  PubMed  Google Scholar 

  2. Hardy RR (2003) Isolation of Ly-1+/CD5+ B cells by cell sorting. Curr Protoc Immunol Chapter 3:Unit 3.5B. doi: 10.1002/ 0471142735. im0305bs55

    Google Scholar 

  3. Tung JW, Mrazek MD, Yang Y, Herzenberg LA, Herzenberg LA (2006) Phenotypically distinct B cell development pathways map to the three B cell lineages in the mouse. Proc Natl Acad Sci U S A 103:6293–6298

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Kasaian MT, Casali P (1993) Autoimmunity-prone B-1 (CD5 B) cells, natural antibodies and self recognition. Autoimmunity 15:315–329

    Article  CAS  PubMed  Google Scholar 

  5. Wen L, Brill-Dashoff J, Shinton SA, Asano M, Hardy RR, Hayakawa K (2005) Evidence of marginal-zone B cell-positive selection in spleen. Immunity 23:297–308

    Article  CAS  PubMed  Google Scholar 

  6. Kroese FG, Ammerlaan WA, Deenen GJ (1992) Location and function of B-cell lineages. Ann N Y Acad Sci 651:44–58

    Article  CAS  PubMed  Google Scholar 

  7. Katz SI, Parker D, Turk JL (1974) B-cell suppression of delayed hypersensitivity reactions. Nature 251:550–551

    Article  CAS  PubMed  Google Scholar 

  8. Neta R, Salvin SB (1974) Specific suppression of delayed hypersensitivity: the possible presence of a suppressor B cell in the regulation of delayed hypersensitivity. J Immunol 113:1716–1725

    CAS  PubMed  Google Scholar 

  9. Mizoguchi A, Bhan AK (2006) A case for regulatory B cells. J Immunol 176:705–710

    Article  CAS  PubMed  Google Scholar 

  10. Lundy SK (2009) Killer B lymphocytes: the evidence and the potential. Inflamm Res 58:345–357

    Article  CAS  PubMed  Google Scholar 

  11. Matsushita T, Horikawa M, Iwata Y, Tedder TF (2010) Regulatory B cells (B10 cells) and regulatory T cells have independent roles in controlling experimental autoimmune encephalomyelitis initiation and late-phase immunopathogenesis. J Immunol 185:2240–2252

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Lo-Man R (2011) Regulatory B cells control dendritic cell functions. Immunotherapy 3(4 Suppl):19–20

    Article  CAS  PubMed  Google Scholar 

  13. Morva A, Lemoine S, Achour A, Pers JO, Youinou P, Jamin C (2012) Maturation and function of human dendritic cells are regulated by B lymphocytes. Blood 119:106–114

    Article  CAS  PubMed  Google Scholar 

  14. Höllsberg P, Batra V, Dressel A, Hafler DA (1996) Induction of anergy in CD8 T cells by B cell presentation of antigen. J Immunol 157:5269–5276

    PubMed  Google Scholar 

  15. Matsushita T, Tedder TF (2011) Identifying regulatory B cells (B10 cells) that produce IL-10 in mice. Methods Mol Biol 677:99–111

    Article  CAS  PubMed  Google Scholar 

  16. Mizoguchi A, Mizoguchi E, Takedatsu H, Blumberg RS, Bhan AK (2002) Chronic intestinal inflammatory condition generates IL-10-producing regulatory B cell subset characterized by CD1d up-regulation. Immunity 16:219–230

    Article  CAS  PubMed  Google Scholar 

  17. Dalwadi H, Wei B, Schrage M, Spicher K, Su TT, Birnbaumer L, Rawlings DJ, Braun J (2003) B cell developmental requirement for the Gαi2 gene. J Immunol 170:1707–1715

    Article  CAS  PubMed  Google Scholar 

  18. Fillatreau S, Sweenie CH, McGeachy MJ, Gray D, Anderton SM (2002) B cells regulate autoimmunity by provision of IL-10. Nat Immunol 3:944–950

    Article  CAS  PubMed  Google Scholar 

  19. Yoshizaki A, Miyagaki T, DiLillo DJ, Matsushita T, Horikawa M, Kountikov EI, Spolski R, Poe JC, Leonard WJ, Tedder TF (2012) Regulatory B cells control T-cell autoimmunity through IL-21-dependent cognate interactions. Nature 491:264–268

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  20. Lenert P, Brummel R, Field H, Ashman RF (2005) TLR-9 activation of marginal zone B cells in lupus mice regulates immunity through increased IL-10 production. J Clin Immunol 25:29–40

    Article  CAS  PubMed  Google Scholar 

  21. Brummel R, Lenert P (2005) Activation of marginal zone B cells from lupus mice with type A(D) CpG-oligodeoxy-nucleotides. J Immunol 174:2429–2434

    Article  CAS  PubMed  Google Scholar 

  22. Mauri C, Gray D, Mushtaq N, Londei M (2003) Prevention of arthritis by interleukin 10-producing B cells. J Exp Med 197:489–501

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  23. Iwata Y, Matsushita T, Horikawa M, Dilillo DJ, Yanaba K, Venturi GM, Szabolcs PM, Bernstein SH, Magro CM, Williams AD, Hall RP, St Clair EW, Tedder TF (2011) Characterization of a rare IL-10-competent B-cell subset in humans that parallels mouse regulatory B10 cells. Blood 117:530–541

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Tian J, Zekzer D, Hanssen L, Lu Y, Olcott A, Kaufman DL (2001) Lipopolysaccharide-activated B cells down-regulate Th1 immunity and prevent autoimmune diabetes in nonobese diabetic mice. J Immunol 167:1081–1089

    Article  CAS  PubMed  Google Scholar 

  25. Flores-Borja F, Bosma A, Ng D, Reddy V, Ehrenstein MR, Isenberg DA, Mauri C (2013) CD19+CD24hiCD38hi B cells maintain regulatory T cells while limiting TH1 and TH17 differentiation. Sci Transl Med 5:173ra23

    Article  PubMed  Google Scholar 

  26. Gonnella PA, Waldner HP, Weiner HL (2001) B cell-deficient (μMT) mice have alterations in the cytokine microenvironment of the gut-associated lymphoid tissue (GALT) and a defect in the low dose mechanism of oral tolerance. J Immunol 166:4456–4464

    Article  CAS  PubMed  Google Scholar 

  27. Parekh VV, Prasad DV, Banerjee PP, Joshi BN, Kumar A, Mishra GC (2003) B cells activated by lipopolysaccharide, but not by anti-Ig and anti-CD40 antibody, induce anergy in CD8+ T cells: role of TGF-β1. J Immunol 170:5897–5911

    Article  CAS  PubMed  Google Scholar 

  28. Singh A, Carson WF IV, Secor ER Jr, Guernsey LA, Flavell RA, Clark RB, Thrall RS, Schramm CM (2008) Regulatory role of B cells in a murine model of allergic airway disease. J Immunol 180:7318–7326

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  29. Natarajan P, Singh A, McNamara JT, Secor ER Jr, Guernsey LA, Thrall RS, Schramm CM (2012) Regulatory B cells from hilar lymph nodes of tolerant mice in a murine model of allergic airway disease are CD5+, express TGF-β, and co-localize with CD4+Foxp3+ T cells. Mucosal Immunol 5:691–701

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  30. Kessel A, Haj T, Peri R, Snir A, Melamed D, Sabo E, Toubi E (2012) Human CD19(+) CD25(high) B regulatory cells suppress proliferation of CD4(+) T cells and enhance Foxp3 and CTLA-4 expression in T-regulatory cells. Autoimmun Rev 11:670–677

    Article  CAS  PubMed  Google Scholar 

  31. Noh J, Lee JH, Noh G, Bang SY, Kim HS, Choi WS, Cho S, Lee SS (2010) Characterisation of allergen-specific responses of IL-10-producing regulatory B cells (Br1) in Cow Milk Allergy. Cell Immunol 264:143–149

    Article  CAS  PubMed  Google Scholar 

  32. Lee JH, Noh J, Noh G, Choi WS, Cho S, Lee SS (2011) Allergen-specific transforming growth factor-β-producing CD19+CD5+ regulatory B-cell (Br3) responses in human late eczematous allergic reactions to cow’s milk. J Interferon Cytokine Res 31:441–449

    Article  CAS  PubMed  Google Scholar 

  33. Ray A, Basu S, Williams CB, Salzman NH, Dittel BN (2012) A novel IL-10-independent regulatory role for B cells in suppressing autoimmunity by maintenance of regulatory T cells via GITR ligand. J Immunol 188:3188–3198

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  34. Minagawa R, Okano S, Tomita Y, Kishihara K, Yamada H, Nomoto K, Shimada M, Maehara Y, Sugimachi K, Yoshikai Y, Nomoto K (2004) The critical role of Fas-Fas ligand interaction in donor-specific transfusion-induced tolerance to H-Y antigen. Transplantation 78:799–806

    Article  CAS  PubMed  Google Scholar 

  35. Lundy SK, Berlin AA, Martens TF, Lukacs NW (2005) Deficiency of regulatory B cells increases allergic airway inflammation. Inflamm Res 54:514–521

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  36. Zhong X, Tumang JR, Gao W, Bai C, Rothstein TL (2007) PD-L2 expression extends beyond dendritic cells/macrophages to B1 cells enriched for V(H)11/V(H)12 and phosphatidylcholine binding. Eur J Immunol 37:2405–2410

    Article  CAS  PubMed  Google Scholar 

  37. Singh AK, Stock P, Akbari O (2011) Role of PD-L1 and PD-L2 in allergic diseases and asthma. Allergy 66:155–162

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  38. Yanaba K, Bouaziz JD, Haas KM, Poe JC, Fujimoto M, Tedder TF (2008) A regulatory B cell subset with a unique CD1dhiCD5+ phenotype controls T cell-dependent inflammatory responses. Immunity 28:639–650

    Article  CAS  PubMed  Google Scholar 

  39. Sonoda KH, Stein-Streilein J (2002) CD1d on antigen-transporting APC and splenic marginal zone B cells promotes NKT cell-dependent tolerance. Eur J Immunol 32:848–857

    Article  CAS  PubMed  Google Scholar 

  40. Wei B, Velazquez P, Turovskaya O, Spricher K, Aranda R, Kronenberg M, Birnbaumer L, Braun J (2005) Mesenteric B cells centrally inhibit CD4+ T cell colitis through interaction with regulatory T cell subsets. Proc Natl Acad Sci U S A 102:2010–2015

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  41. Dufour FD, Silveira PA, Baxter AG (2008) Interactions between B-lymphocytes and type 1 NKT cells in autoimmune diabetes. J Immunotoxicol 5:249–257

    Article  CAS  PubMed  Google Scholar 

  42. Schramm CM, Puddington L, Wu C, Guernsey L, Gharaee-Kermani M, Phan SH, Thrall RS (2004) Chronic inhaled ovalbumin exposure induces antigen-dependent but not antigen-specific inhalational tolerance in a murine model of allergic airway disease. Am J Pathol 164:295–304

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  43. Singh A, Thrall RS, Guernsey LA, Carson WF IV, Secor ER Jr, Cone RE, Chattopadhyay S, Rajan TV, Schramm CM (2008) Subcutaneous late phase responses are augmented during local inhalational tolerance in a murine asthma model. Immunol Cell Biol 86:535–538

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  44. Carson WF IV, Guernsey LA, Singh A, Vella AT, Schramm CM, Thrall RS (2008) Accumulation of regulatory T cells in the local draining lymph nodes of the lung correlates with the spontaneous resolution of a chronic model of murine asthma. Int Arch Allergy Immunol 145:231–243

    Article  PubMed Central  PubMed  Google Scholar 

  45. McNamara JT, Schramm CM, Singh A, Secor ER Jr, Guernsey LA, Lefrançois L, Thrall RS (2012) Phenotypic changes to the endogenous antigen-specific CD8+ T cell response correlate with the development and resolution of allergic airway disease. Am J Pathol 180:1991–2000

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Acknowledgments

This work was supported by grants from the National Institutes of Health R21–AI079533 and R01–HL43573.

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Authors and Affiliations

  1. Department of Immunology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA

    Prabitha Natarajan & Linda A. Guernsey

  2. Department of Pediatrics, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA

    Craig M. Schramm M.D.

Authors
  1. Prabitha Natarajan
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  2. Linda A. Guernsey
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  3. Craig M. Schramm M.D.
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Corresponding author

Correspondence to Craig M. Schramm M.D. .

Editor information

Editors and Affiliations

  1. Department of Medical and Biological Sciences, University of Udine, Udine, Italy

    Gaetano Vitale

  2. Department of Medical and Biological Sciences, University of Udine, Udine, Italy

    Francesca Mion

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Natarajan, P., Guernsey, L.A., Schramm, C.M. (2014). Regulatory B Cells in Allergic Airways Disease and Asthma. In: Vitale, G., Mion, F. (eds) Regulatory B Cells. Methods in Molecular Biology, vol 1190. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1161-5_15

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  • DOI: https://doi.org/10.1007/978-1-4939-1161-5_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1160-8

  • Online ISBN: 978-1-4939-1161-5

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