Flow Cytometric Analysis of Circulating Follicular Helper T (Tfh) and Follicular Regulatory T (Tfr) Populations in Human Blood

  • Yunbo Wei
  • Jinhong Feng
  • Zhaohua Hou
  • Xin Maggie Wang
  • Di Yu
Part of the Methods in Molecular Biology book series (MIMB, volume 1291)

Abstract

Follicular helper T (Tfh) cells are the helper T-cell subset that localizes in germinal centers within secondary lymphoid organs. They support B cells to produce high-affinity antibodies and generate B-cell memory. By contrast, follicular regulatory T (Tfr) cells localize in germinal centers to suppress B-cell responses. Coordinately regulating antibody responses, Tfh and Tfr cells play a critical role for vaccination, infectious disease control, and the development of autoimmune diseases. Although bona fide Tfh and Tfr cells are hardly detected in human blood, circulating Tfh and Tfr memory cells in blood can be used to investigate their functions in health and disease. We have developed a comprehensive flow cytometric analysis to define different circulating Tfh and Tfr populations within CXCR5+ CD4+ T-cell population in human blood, based on numerous cell surface markers including CD25, CD127, CCR7, PD-1, CXCR3, and CCR6.

Key words

Tfh Tfr Human blood Flow cytometry CXCR5 

References

  1. 1.
    Yu D, Vinuesa CG (2010) The elusive identity of T follicular helper cells. Trends Immunol 31(10):377–383. doi:10.1016/j.it.2010.07.001 CrossRefPubMedGoogle Scholar
  2. 2.
    Crotty S (2011) Follicular helper CD4 T cells (TFH). Annu Rev Immunol 29:621–663. doi:10.1146/annurev-immunol-031210-101400 CrossRefPubMedGoogle Scholar
  3. 3.
    Yu D, Vinuesa CG (2010) Multiple checkpoints keep follicular helper T cells under control to prevent autoimmunity. Cell Mol Immunol 7(3):198–203. doi:10.1038/cmi.2010.18 CrossRefPubMedCentralPubMedGoogle Scholar
  4. 4.
    Tangye SG, Ma CS, Brink R, Deenick EK (2013) The good, the bad and the ugly—TFH cells in human health and disease. Nat Rev Immunol 13(6):412–426. doi:10.1038/nri3447 CrossRefPubMedGoogle Scholar
  5. 5.
    He J, Tsai LM, Leong YA, Hu X, Ma CS, Chevalier N, Sun X, Vandenberg K, Rockman S, Ding Y, Zhu L, Wei W, Wang C, Karnowski A, Belz G, Ghali J, Cook MC, Riminton DS, Veillette A, Schwartzberg PL, MacKay F, Brink R, Tangye SG, Vinuesa CG, Mackay CR, Li Z, Yu D (2013) Circulating precursor CCR7loPD-1hi CXCR5+ CD4 T cells identify active follicular T helper programs in health and disease. Immunity 39(4):770–781. doi:10.1016/j.immuni.2013.09.007 CrossRefPubMedGoogle Scholar
  6. 6.
    Tsai LM, Yu D (2014) Follicular helper T-cell memory: establishing new frontiers during antibody response. Immunol Cell Biol 92(1):57–63. doi:10.1038/icb.2013.68 CrossRefPubMedGoogle Scholar
  7. 7.
    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. doi:10.1038/nm.2426 CrossRefPubMedCentralPubMedGoogle Scholar
  8. 8.
    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. doi:10.1038/nm.2425 CrossRefPubMedCentralPubMedGoogle Scholar
  9. 9.
    Wollenberg I, Agua-Doce A, Hernandez A, Almeida C, Oliveira VG, Faro J, Graca L (2011) Regulation of the germinal center reaction by Foxp3+ follicular regulatory T cells. J Immunol 187(9):4553–4560. doi:10.4049/jimmunol.1101328 CrossRefPubMedGoogle Scholar
  10. 10.
    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. doi:10.1038/ni.2496 CrossRefPubMedCentralPubMedGoogle Scholar
  11. 11.
    Simpson N, Gatenby PA, Wilson A, Malik S, Fulcher DA, Tangye SG, Manku H, Vyse TJ, Roncador G, Huttley GA, Goodnow CC, Vinuesa CG, Cook MC (2010) Expansion of circulating T cells resembling follicular helper T cells is a fixed phenotype that identifies a subset of severe systemic lupus erythematosus. Arthritis Rheum 62(1):234–244. doi:10.1002/art.25032 CrossRefPubMedGoogle Scholar
  12. 12.
    Morita R, Schmitt N, Bentebibel SE, Ranganathan R, Bourdery L, Zurawski G, Foucat E, Dullaers M, Oh S, Sabzghabaei N, Lavecchio EM, Punaro M, Pascual V, Banchereau J, Ueno H (2011) Human blood CXCR5(+)CD4(+) T cells are counterparts of T follicular cells and contain specific subsets that differentially support antibody secretion. Immunity 34(1):108–121. doi:10.1016/j.immuni.2010.12.012 CrossRefPubMedCentralPubMedGoogle Scholar
  13. 13.
    Bentebibel SE, Lopez S, Obermoser G, Schmitt N, Mueller C, Harrod C, Flano E, Mejias A, Albrecht RA, Blankenship D, Xu H, Pascual V, Banchereau J, Garcia-Sastre A, Palucka AK, Ramilo O, Ueno H (2013) Induction of ICOS + CXCR3 + CXCR5+ TH cells correlates with antibody responses to influenza vaccination. Sci Transl Med 5(176):176ra–132. doi:10.1126/scitranslmed.3005191 Google Scholar
  14. 14.
    Locci M, Havenar-Daughton C, Landais E, Wu J, Kroenke MA, Arlehamn CL, Su LF, Cubas R, Davis MM, Sette A, Haddad EK, International AVIPCPI, Poignard P, Crotty S (2013) Human circulating PD-(+)1CXCR3(-)CXCR5(+) memory Tfh cells are highly functional and correlate with broadly neutralizing HIV antibody responses. Immunity 39(4):758–769. doi:10.1016/j.immuni.2013.08.031 CrossRefPubMedCentralPubMedGoogle Scholar
  15. 15.
    Liu W, Putnam AL, Xu-Yu Z, Szot GL, Lee MR, Zhu S, Gottlieb PA, Kapranov P, Gingeras TR, Fazekas de St Groth B, Clayberger C, Soper DM, Ziegler SF, Bluestone JA (2006) CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells. J Exp Med 203(7):1701–1711. doi:10.1084/jem.20060772 CrossRefPubMedCentralPubMedGoogle Scholar
  16. 16.
    Seddiki N, Santner-Nanan B, Martinson J, Zaunders J, Sasson S, Landay A, Solomon M, Selby W, Alexander SI, Nanan R, Kelleher A, Fazekas de St Groth B (2006) Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells. J Exp Med 203(7):1693–1700. doi:10.1084/jem.20060468 CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Schmitt N, Ueno H (2013) Blood Tfh cells come with colors. Immunity 39(4):629–630. doi:10.1016/j.immuni.2013.09.011 CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Yunbo Wei
    • 1
    • 2
  • Jinhong Feng
    • 1
  • Zhaohua Hou
    • 1
  • Xin Maggie Wang
    • 3
  • Di Yu
    • 1
    • 4
  1. 1.Laboratory of Immunology for Environment and Health, Shandong Analysis and Test CenterShandong Academy of SciencesJinanChina
  2. 2.Molecular Immunomodulation Laboratory, School of Biomedical SciencesMonash UniversityMelbourneAustralia
  3. 3.Flow Cytometry CentreWestmead Millennium InstituteWestmeadAustralia
  4. 4.Molecular Immunomodulation Laboratory, School of Biomedical SciencesMonash UniversityMelbourneAustralia

Personalised recommendations