Abstract
T Follicular helper (Tfh) cells stimulate, whereas T follicular regulatory (Tfr) cells inhibit, effector B cell responses. Although new tools have been developed to assess the functional roles of Tfh and Tfr cells in vivo, methods to assess mechanisms have been limited. One such limitation has been the ability of in vitro functional assays to recapitulate robust germinal center-like responses. Although previous in vitro Tfh-mediated and Tfr-suppressed assays to assess antibody regulation have been developed, these classically have relied on polyclonal stimulation. To understand Tfh and Tfr cell functionality, more robust assays that utilize specific antigen are needed. Here we describe an in vitro approach for sensitively and quantitatively assessing the capacity of Tfh and Tfr cells to regulate B cell responses in an antigen-driven system. These assays allow the study of Tfh and Tfr cells in specific disease contexts, such as IgG production after vaccination or IgE responses during allergic airway disease.
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
Crotty S (2014) T follicular helper cell differentiation, function, and roles in disease. Immunity 41(4):529–542. https://doi.org/10.1016/j.immuni.2014.10.004
Sage PT, Sharpe AH (2016) T follicular regulatory cells. Immunol Rev 271(1):246–259. https://doi.org/10.1111/imr.12411
Fonseca VR, Ribeiro F, Graca L (2019) T follicular regulatory (Tfr) cells: dissecting the complexity of Tfr-cell compartments. Immunol Rev 288(1):112–127. https://doi.org/10.1111/imr.12739
Sage PT, Sharpe AH (2020) The multifaceted functions of follicular regulatory T cells. Curr Opin Immunol 67:68–74. https://doi.org/10.1016/j.coi.2020.10.009
Clement RL, Daccache J, Mohammed MT, Diallo A, Blazar BR, Kuchroo VK, Lovitch SB, Sharpe AH, Sage PT (2019) Follicular regulatory T cells control humoral and allergic immunity by restraining early B cell responses. Nat Immunol 20(10):1360–1371. https://doi.org/10.1038/s41590-019-0472-4
Wu H, Chen Y, Liu H, Xu LL, Teuscher P, Wang S, Lu S, Dent AL (2016) Follicular regulatory T cells repress cytokine production by follicular helper T cells and optimize IgG responses in mice. Eur J Immunol 46(5):1152–1161. https://doi.org/10.1002/eji.201546094
Fu W, Liu X, Lin X, Feng H, Sun L, Li S, Chen H, Tang H, Lu L, Jin W, Dong C (2018) Deficiency in T follicular regulatory cells promotes autoimmunity. J Exp Med 215(3):815–825. https://doi.org/10.1084/jem.20170901
Sage PT, Francisco LM, Carman CV, Sharpe AH (2013) The receptor PD-1 controls follicular regulatory T cells in the lymph nodes and blood. Nat Immunol 14(2):152–161
Sage PT, Schildberg FA, Sobel RA, Kuchroo VK, Freeman GJ, Sharpe AH (2018) Dendritic cell PD-L1 limits autoimmunity and follicular T cell differentiation and function. J Immunol 200(8):2592–2602. https://doi.org/10.4049/jimmunol.1701231
Sage PT, Paterson AM, Lovitch SB, Sharpe AH (2014) The coinhibitory receptor ctla-4 controls B cell responses by modulating T follicular helper, T follicular regulatory, and T regulatory cells. Immunity 41(6):1026–1039. https://doi.org/10.1016/j.immuni.2014.12.005
Sage PT, Ron-Harel N, Juneja VR, Sen DR, Maleri S, Sungnak W, Kuchroo VK, Haining WN, Chevrier N, Haigis M, Sharpe AH (2016) Suppression by TFR cells leads to durable and selective inhibition of B cell effector function. Nat Immunol 17(12):1436–1446. https://doi.org/10.1038/ni.3578
Hou S, Clement RL, Diallo A, Blazar BR, Rudensky AY, Sharpe AH, Sage PT (2019) FoxP3 and Ezh2 regulate Tfr cell suppressive function and transcriptional program. J Exp Med 216(3):605–620. https://doi.org/10.1084/jem.20181134
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Clement, R.L., Hanson, B.L., Sage, P.T. (2022). Tfh-Mediated and Tfr-Suppressed Antigen-Driven IgG and IgE Assays. In: Graca, L. (eds) T-Follicular Helper Cells. Methods in Molecular Biology, vol 2380. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1736-6_15
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1736-6_15
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1735-9
Online ISBN: 978-1-0716-1736-6
eBook Packages: Springer Protocols