Abstract
Follicular regulatory T (TFR) cells, a subset of Foxp3+ regulatory T cells (Tregs), play an essential role in the regulation of germinal center (GC) response and humoral self-tolerance. Although it is generally accepted that TFR cells suppress GC antibody responses mediated by follicular helper T (TFH) cells and B cells, the helper activity of TFR cells on GC responses has also been recently reported. Because of this, it is critical to develop specific assays that are able to precisely assess TFR cell function, particularly its in vivo activity, independent of differentiation and other Tregs. Here we describe an adoptive transfer approach in conjugation with flow cytometry and ELISA to evaluate the TFR cell function on TFH, B cells, and antibody response in vivo.
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References
Leavenworth JW, Verbinnen B, Yin J, Huang H, Cantor H (2015) A p85alpha-osteopontin axis couples the receptor ICOS to sustained Bcl-6 expression by follicular helper and regulatory T cells. Nat Immunol 16(1):96–106. https://doi.org/10.1038/ni.3050
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
Linterman MA, Pierson W, Lee SK, Kallies A, Kawamoto S, Rayner TF, Srivastava M, Divekar DP, Beaton L, Hogan JJ, Fagarasan S, Liston A, Smith KG, Vinuesa CG (2011) Foxp3+ follicular regulatory T cells control the germinal center response. Nat Med 17(8):975–982. https://doi.org/10.1038/nm.2425
Chung Y, Tanaka S, Chu F, Nurieva RI, Martinez GJ, Rawal S, Wang YH, Lim H, Reynolds JM, Zhou XH, Fan HM, Liu ZM, Neelapu SS, Dong C (2011) Follicular regulatory T cells expressing Foxp3 and Bcl-6 suppress germinal center reactions. Nat Med 17(8):983–988. https://doi.org/10.1038/nm.2426
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
Shen E, Rabe H, Luo L, Wang L, Wang Q, Yin J, Yang X, Liu W, Sido JM, Nakagawa H, Ao L, Kim HJ, Cantor H, Leavenworth JW (2019) Control of germinal center localization and lineage stability of follicular regulatory T cells by the Blimp1 transcription factor. Cell Rep 29(7):1848–1861. e1846. https://doi.org/10.1016/j.celrep.2019.10.012
Laidlaw BJ, Lu Y, Amezquita RA, Weinstein JS, Vander Heiden JA, Gupta NT, Kleinstein SH, Kaech SM, Craft J (2017) Interleukin-10 from CD4(+) follicular regulatory T cells promotes the germinal center response. Sci Immunol 2(16). https://doi.org/10.1126/sciimmunol.aan4767
Xie MM, Chen Q, Liu H, Yang K, Koh B, Wu H, Maleki SJ, Hurlburt BK, Cook-Mills J, Kaplan MH, Dent AL (2020) T follicular regulatory cells and IL-10 promote food antigen-specific IgE. J Clin Invest 130(7):3820–3832. https://doi.org/10.1172/JCI132249
Shen E, Wang Q, Rabe H, Liu W, Cantor H, Leavenworth JW (2018) Chromatin remodeling by the NuRD complex regulates development of follicular helper and regulatory T cells. Proc Natl Acad Sci USA 115(26):6780–6785. https://doi.org/10.1073/pnas.1805239115
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. https://doi.org/10.1038/ni.2496
Takahashi Y, Dutta PR, Cerasoli DM, Kelsoe G (1998) In situ studies of the primary immune response to (4-hydroxy-3-nitrophenyl)acetyl. V. Affinity maturation develops in two stages of clonal selection. J Exp Med 187(6):885–895. https://doi.org/10.1084/jem.187.6.885
Leavenworth JW, Wang X, Wenander CS, Spee P, Cantor H (2011) Mobilization of natural killer cells inhibits development of collagen-induced arthritis. Proc Natl Acad Sci USA 108(35):14584–14589. https://doi.org/10.1073/pnas.1112188108
Leavenworth JW, Tang X, Kim HJ, Wang X, Cantor H (2013) Amelioration of arthritis through mobilization of peptide-specific CD8+ regulatory T cells. J Clin Invest 123(3):1382–1389. https://doi.org/10.1172/JCI66938
Kang SG, Liu WH, Lu P, Jin HY, Lim HW, Shepherd J, Fremgen D, Verdin E, Oldstone MB, Qi H, Teijaro JR, Xiao C (2013) MicroRNAs of the miR-17 approximately 92 family are critical regulators of T(FH) differentiation. Nat Immunol 14(8):849–857. https://doi.org/10.1038/ni.2648
Acknowledgments
This work was supported by NIH grant R01AI148711 and the University of Alabama at Birmingham faculty start-up funds to J.W.L. J.W.L is also supported by DoD W81XWH-18-1-0315.
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Luo, L., Leavenworth, J.W. (2023). Analysis of the In Vivo Function of Follicular Regulatory T (TFR) Cells in the Regulation of Antibody Response. In: Ono, M. (eds) Regulatory T-Cells. Methods in Molecular Biology, vol 2559. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2647-4_1
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DOI: https://doi.org/10.1007/978-1-0716-2647-4_1
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