Abstract
Regulatory B10 cells represent a rare CD1dhiCD5+ IL-10-secreting B cell subset in mice which is induced to produce IL-10 after 5 h of in vitro stimulation with a combination of B cell mitogen and chemical stimulants. Although B10 cells only constitute 1–2 % of splenic CD19+ B cells, they play important roles in controlling T cell-mediated immune responses in an antigen-specific and IL-10-dependent manner. The regulatory effects of B10 cells have been demonstrated in multiple mouse models of inflammatory and autoimmune diseases. Herein, we described current methods for identification and purification of CD1dhiCD5+ B10 cells.
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References
Katz SI, Parker D, Turk JL (1974) B-cell suppression of delayed hypersensitivity reactions. Nature 251:550–551
Wolf SD, Dittel BN, Hardardottir F, Janeway CA Jr (1996) Experimental autoimmune encephalomyelitis induction in genetically B cell-deficient mice. J Exp Med 184:2271–2278
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
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
Mauri C, Gray D, Mushtaq N, Londei M (2003) Prevention of arthritis by interleukin 10-producing B cells. J Exp Med 197:489–501
Mauri C, Bosma A (2012) Immune regulatory function of B cells. Annu Rev Immunol 30: 221–241
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 upregulation. Immunity 16:219–230
Hussain S, Delovitch TL (2007) Intravenous transfusion of BCR-activated B cells protects NOD mice from type 1 diabetes in an IL-10-dependent manner. J Immunol 179:7225–7232
Evans JG, Chavez-Rueda KA, Eddaoudi A, Meyer-Bahlburg A, Rawlings DJ, Ehrenstein MR et al (2007) Novel suppressive function of transitional 2 B cells in experimental arthritis. J Immunol 178:7868–7878
Harris DP, Haynes L, Sayles PC, Duso DK, Eaton SM, Lepak NM et al (2000) Reciprocal regulation of polarized cytokine production by effector B and T cells. Nat Immunol 1:475–482
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
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
Yanaba K, Bouaziz JD, Matsushita T, Tsubata T, Tedder TF (2009) The development and function of regulatory B cells expressing IL-10 (B10 cells) requires antigen receptor diversity and TLR signals. J Immunol 182:7459–7472
Yoshizaki A, Miyagaki T, DiLillo DJ, Matsushita T, Horikawa M, Kountikov EI et al (2012) Regulatory B cells control T-cell autoimmunity through IL-21-dependent cognate interactions. Nature 491:264–268
Hong C, Zhang T, Gao XM (2013) Recombinant murine calreticulin fragment 39-272 expands CD1d(hi)CD5+ IL-10-secreting B cells that modulate experimental autoimmune encephalomyelitis in C57BL/6 mice. Mol Immunol 55:237–246
Acknowledgements
The authors are supported by grants from PCSIRT (IRT1075), National Key Basic Research Programs (2010CB529102), and National Foundation of Nature Science (31370908/31070781/31100633) of China.
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Hong, C., Gao, XM. (2014). Purification and Immunophenotypic Characterization of Murine B10 B Cells. 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_3
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DOI: https://doi.org/10.1007/978-1-4939-1161-5_3
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