Imbalance of circulating Tfr/Tfh ratio in patients with rheumatoid arthritis
Follicular helper T(Tfh) cells and follicular regulatory T(Tfr) cells are critical for the development and maintenance of germinal center and humoral immune responses. Accumulating evidence has demonstrated that the dysregulation of either Tfh or Tfr cells contributes to the pathogenesis of autoimmune diseases. The aim of this study was to examine the numbers of Tfh and Tfr cells in patients with rheumatoid arthritis (RA). Twenty-four patients with RA patients and 20 health controls (HCs) were enrolled in this study. We analyzed the numbers of Tfh (CD4+ CXCR5+ PD-1hi) cells and Tfr (CD4+ CXCR5+CD127lo) cells in 24 RA patients via flow cytometry. The level of the soluble PD-1 and its ligands (sPD-L1 and sPDL-2) were examined by ELISA. Flow cytometry revealed that both circulating Tfh and Tfr cells were increased in RA patients compared with HCs. More importantly, the ratio of Tfr/Tfh was decreased, indicating a disruption of the balance between Tfh and Tfr. The Tfr/Tfh ratio was inversely correlated with level of serum CRP, ESR, RF, anti-CCP, IgG and DAS28 index. We also found that the serum level of sPD-1 was significantly elevated in the RA patients, which was positively correlated with CRP, ESR and the number of Tfh cells. These results indicate that an imbalance of circulating Tfr and Tfh cells may be involved in the immunopathogenesis of RA and may provide novel insight for the development of RA therapies.
KeywordsRheumatoid arthritis Follicular helper T cell(Tfh) Follicular regulatory T cell(Tfr) Tfr/Tfh PD-1
Circulating follicular helper T
Circulating follicular regulatory T
Enzyme‐linked immunosorbent assay
Erythrocyte sedimentation rate
CXC chemokine receptor 5
Cyclic citrullinated peptide
Disease activity score in 28 joints
Disease-modifying antirheumatic drug
This work was supported by the following grants: foundation from clinical medical research center of Shenyang City (18_009-4-03 to PT.Y.), foundation from the Major State Research Development Program of Liaoning, China (No. 2017225024 to PT.Y.), foundation from the Project for Construction of Major Discipline Platform in Universities of Liaoning province, China (2017001 to PT.Y.), the Program of the Distinguished Professor of Liaoning Province, Rheumatology (2017 to PT.Y.).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
The studies have been approved by the ethics committee of the first affiliated hospital of China Medical University. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
- 5.Crotty S. Follicular helper CD4 T cells (TFH). Annu Rev Immunol. 2011;29:621–63. https://doi.org/10.1146/annurev-immunol-031210-101400.CrossRefPubMedGoogle Scholar
- 12.He J, Tsai LM, Leong YA, Hu X, Ma CS, Chevalier N, et al. Circulating precursor CCR7(lo)PD-1(hi) CXCR5(+) CD4(+) T cells indicate Tfh cell activity and promote antibody responses upon antigen reexposure. Immunity. 2013;39(4):770–81. https://doi.org/10.1016/j.immuni.2013.09.007.CrossRefPubMedGoogle Scholar
- 15.Xu B, Wang S, Zhou M, Huang Y, Fu R, Guo C, et al. The ratio of circulating follicular T helper cell to follicular T regulatory cell is correlated with disease activity in systemic lupus erythematosus. Clin Immunol. 2017;183:46–53. https://doi.org/10.1016/j.clim.2017.07.004.CrossRefPubMedPubMedCentralGoogle Scholar
- 19.Arroyo-Villa I, Bautista-Caro MB, Balsa A, Aguado-Acin P, Bonilla-Hernan MG, Plasencia C, et al. Constitutively altered frequencies of circulating follicullar helper T cell counterparts and their subsets in rheumatoid arthritis. Arthritis Res Therapy. 2014;16(6):500. https://doi.org/10.1186/s13075-014-0500-6.CrossRefGoogle Scholar
- 20.Wang J, Shan Y, Jiang Z, Feng J, Li C, Ma L, et al. High frequencies of activated B cells and T follicular helper cells are correlated with disease activity in patients with new-onset rheumatoid arthritis. Clin Exp Immunol. 2013;174(2):212–20. https://doi.org/10.1111/cei.12162.CrossRefPubMedPubMedCentralGoogle Scholar
- 22.Chakera A, Bennett SC, Morteau O, Bowness P, Luqmani RA, Cornall RJ. The phenotype of circulating follicular-helper T cells in patients with rheumatoid arthritis defines CD200 as a potential therapeutic target. Clin Dev Immunol. 2012;2012:948218. https://doi.org/10.1155/2012/948218.CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Aletaha D, Neogi T, Silman AJ, Funovits J, Felson DT, Bingham CO 3rd, et al. Rheumatoid arthritis classification criteria: an American College of Rheumatology/European league against rheumatism collaborative initiative. Arthritis Rheum. 2010;62(9):2569–81. https://doi.org/10.1002/art.27584.CrossRefPubMedGoogle Scholar
- 25.Prevoo ML, van ‘t Hof MA, Kuper HH, van Leeuwen MA, van de Putte LB, van de Riel PL. Modified disease activity scores that include twenty-eight-joint counts. Development and validation in a prospective longitudinal study of patients with rheumatoid arthritis. Arthritis Rheum. 1995;38(1):44–8.CrossRefGoogle Scholar
- 26.Simpson N, Gatenby PA, Wilson A, Malik S, Fulcher DA, Tangye SG, et al. 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. 2010;62(1):234–44. https://doi.org/10.1002/art.25032.CrossRefPubMedGoogle Scholar
- 33.Seddiki N, Santner-Nanan B, Martinson J, Zaunders J, Sasson S, Landay A, et al. Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells. J Exp Med. 2006;203(7):1693–700. https://doi.org/10.1084/jem.20060468.CrossRefPubMedPubMedCentralGoogle Scholar
- 34.Hartigan-O’Connor DJ, Poon C, Sinclair E, McCune JM. Human CD4+ regulatory T cells express lower levels of the IL-7 receptor alpha chain (CD127), allowing consistent identification and sorting of live cells. J Immunol Methods. 2007;319(1–2):41–52. https://doi.org/10.1016/j.jim.2006.10.008.CrossRefPubMedGoogle Scholar
- 35.Morita R, Schmitt N, Bentebibel SE, Ranganathan R, Bourdery L, Zurawski G, et al. Human blood CXCR5(+)CD4(+) T cells are counterparts of T follicular cells and contain specific subsets that differentially support antibody secretion. Immunity. 2011;34(1):108–21. https://doi.org/10.1016/j.immuni.2010.12.012.CrossRefPubMedPubMedCentralGoogle Scholar