Abstract
The analysis of human B cell populations of the blood relies on the expression of surface markers, mainly CD19, CD24, CD38, and CD27. According to these surface markers, three main B cell subsets can be identified in the blood: immature transitional B cells (CD19+CD24highCD38high), naïve B cells (CD19+CD24intCD38int) that have not encountered an antigen, and memory B cells (CD19+CD27+). To date, human B cells with regulatory functions have been essentially described within the CD24highCD38high transitional B cell subset. CD24highCD38high transitional B cells are able to produce interleukin 10 (IL-10) and to regulate in vitro Th1 and Th17 CD4+ T cell activation. Here, we provide the methods to analyze and purify the CD24highCD38high transitional B cell subset for further in vitro experiments. We also provide a reliable method to detect B cell IL-10 production using intracellular cytokine staining.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bouaziz JD, Yanaba K, Venturi GM, Wang Y, Tisch RM, Poe JC, Tedder TF (2007) Therapeutic B cell depletion impairs adaptive and autoreactive CD4+ T cell activation in mice. Proc Natl Acad Sci U S A 104:20882–20887
Bouaziz JD, Yanaba K, Tedder TF (2008) Regulatory B cells as inhibitors of immune responses and inflammation. Immunol Rev 224:201–214
Bouaziz JD, Calbo S, Maho-Vaillant M, Saussine A, Bagot M, Bensussan A, Musette P (2010) IL-10 produced by activated human B cells regulates CD4(+) T-cell activation in vitro. Eur J Immunol 40:2686–2691
Blair PA, Noreña LY, Flores-Borja F, Rawlings DJ, Isenberg DA, Ehrenstein MR, Mauri C (2010) CD19(+)CD24(hi)CD38(hi) B cells exhibit regulatory capacity in healthy individuals but are functionally impaired in systemic Lupus Erythematosus patients. Immunity 32:129–140
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 (2010) Characterization of a rare IL-10-competent B-cell subset in humans that parallels mouse regulatory B10 cells. Blood 117:530–541
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 TH2 differentiation. Sci Transl Med 5: 173ra23
Li X, Zhong H, Bao W, Boulad N, Evangelista J, Haider MA, Bussel J, Yazdanbakhsh K (2012) Defective regulatory B-cell compartment in patients with immune thrombocytopenia. Blood 120:3318–3325
Wilde B, Thewissen M, Damoiseaux J, Knippenberg S, Hilhorst M, van Paassen P, Witzke O, Cohen Tervaert JW (2013) Regulatory B cells in ANCA-associated vasculitis. Ann Rheum Dis 72:1416–1419
Saussine A, Tazi A, Feuillet S, Rybojad M, Juillard C, Bergeron A, Dessirier V, Bouhidel F, Janin A, Bensussan A, Bagot M, Bouaziz JD (2012) Active chronic sarcoidosis is characterized by increased transitional blood B cells, increased IL-10-producing regulatory B cells and high BAFF levels. PLoS One 7:e43588
Palanichamy A, Barnard J, Zheng B, Owen T, Quach T, Wei C, Looney RJ, Sanz I, Anolik JH (2009) Novel human transitional B cell populations revealed by B cell depletion therapy. J Immunol 182:5982–5993
Colliou N, Picard D, Caillot F, Calbo S, Le Corre S, Lim A et al (2013) Long-term remissions of severe pemphigus after rituximab therapy are associated with prolonged failure of desmoglein B cell response. Sci Transl Med 5: 175ra30
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
de Masson, A., Le Buanec, H., Bouaziz, JD. (2014). Purification and Immunophenotypic Characterization of Human B Cells with Regulatory Functions. 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_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1161-5_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1160-8
Online ISBN: 978-1-4939-1161-5
eBook Packages: Springer Protocols