Abstract
Regulatory B cells do not constitute a distinct cell lineage because no unique marker or set of markers can exclusively identify neither murine nor human regulatory B cells, and efficient IL-10 production is their only known distinguishing feature. After purification of IL-10-secreting B cells, one may want to characterize them by analyzing their gene expression profile. This goal can be achieved by using different technologies: RT-qPCR, microarrays, Nanostring’s nCounter technology, Biomark HD are techniques that will allow you to analyze their gene expression, whether in a targeted (RT-qPCR), extended but targeted (Nanostring’s nCounter technology, Biomark HD) or exhaustive (Microarray) way. Aim of this chapter is the description of these techniques in the view of their application to the study and characterization of regulatory B cells.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Chattopadhyay PK, Gierahn TM, Roederer M, Love JC (2014) Single-cell technologies for monitoring immune systems. Nat Immunol 15(2):128–135. https://doi.org/10.1038/ni.2796
Rincon-Arevalo H, Sanchez-Parra CC, Castano D, Yassin L, Vasquez G (2016) Regulatory B cells and mechanisms. Int Rev Immunol 35(2):156–176. https://doi.org/10.3109/08830185.2015.1015719
Rosser EC, Mauri C (2015) Regulatory B cells: origin, phenotype, and function. Immunity 42(4):607–612. https://doi.org/10.1016/j.immuni.2015.04.005
Zheng GX, Terry JM, Belgrader P, Ryvkin P, Bent ZW, Wilson R, Ziraldo SB, Wheeler TD, McDermott GP, Zhu J, Gregory MT, Shuga J, Montesclaros L, Underwood JG, Masquelier DA, Nishimura SY, Schnall-Levin M, Wyatt PW, Hindson CM, Bharadwaj R, Wong A, Ness KD, Beppu LW, Deeg HJ, McFarland C, Loeb KR, Valente WJ, Ericson NG, Stevens EA, Radich JP, Mikkelsen TS, Hindson BJ, Bielas JH (2017) Massively parallel digital transcriptional profiling of single cells. Nat Commun 8:14049. https://doi.org/10.1038/ncomms14049
Shalek AK, Satija R, Adiconis X, Gertner RS, Gaublomme JT, Raychowdhury R, Schwartz S, Yosef N, Malboeuf C, Lu D, Trombetta JJ, Gennert D, Gnirke A, Goren A, Hacohen N, Levin JZ, Park H, Regev A (2013) Single-cell transcriptomics reveals bimodality in expression and splicing in immune cells. Nature 498(7453):236–240. https://doi.org/10.1038/nature12172
Berkani N, Joly P, Golinski ML, Colliou N, Lim A, Larbi A, Riou G, Caillot F, Bernard P, Bedane C, Delaporte E, Chaby G, Dompmartin A, Hertl M, Calbo S, Musette P (2019) B-cell depletion induces a shift in self antigen specific B-cell repertoire and cytokine pattern in patients with bullous pemphigoid. Sci Rep 9(1):3525. https://doi.org/10.1038/s41598-019-40203-7
Hebert V, Petit M, Maho-Vaillant M, Golinski ML, Riou G, Derambure C, Boyer O, Joly P, Calbo S (2019) Modifications of the Transcriptomic profile of autoreactive B cells from pemphigus patients after treatment with rituximab or a standard corticosteroid regimen. Front Immunol 10:1794. https://doi.org/10.3389/fimmu.2019.01794
Golinski ML, Demeules M, Derambure C, Riou G, Maho-Vaillant M, Boyer O, Joly P, Calbo S (2020) CD11c(+) B cells are mainly memory cells, precursors of antibody secreting cells in healthy donors. Front Immunol 11:32. https://doi.org/10.3389/fimmu.2020.00032
Lin W, Cerny D, Chua E, Duan K, Yi JT, Shadan NB, Lum J, Maho-Vaillant M, Zolezzi F, Wong SC, Larbi A, Fink K, Musette P, Poidinger M, Calbo S (2014) Human regulatory B cells combine phenotypic and genetic hallmarks with a distinct differentiation fate. J Immunol 193(5):2258–2266. https://doi.org/10.4049/jimmunol.1303214
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Maho-Vaillant, M., Calbo, S. (2021). Methods to Study the Transcriptome of Regulatory B Cells. In: Mion, F., Tonon, S. (eds) Regulatory B Cells. Methods in Molecular Biology, vol 2270. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1237-8_16
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1237-8_16
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1236-1
Online ISBN: 978-1-0716-1237-8
eBook Packages: Springer Protocols