Analysis of PD-1 and Tim-3 expression on CD4+ T cells of patients with rheumatoid arthritis; negative association with DAS28
Expression of T cell immunoglobulin and mucin-domain containing-3 (Tim-3) and programmed cell death-1 (PD-1) was studied on CD4+ T cells of patients with rheumatoid arthritis (RA). Association of Tim-3 and PD-1 expression with disease activity of RA patients was also addressed. A total of 37 RA patients and 31 sex- and age-matched healthy controls were included in this study. Disease activity of RA patients was determined by Disease Activity Score of 28 joints scoring system (DAS28). A three-color flow cytometry method was applied to determine the frequency of Tim-3+/PD-1+/CD4+ T cells. To measure the cytokine production, peripheral blood mononuclear cells (PBMCs) were stimulated with PMA/ionomycin. Concentrations of IL-17, IL-10, IFN-γ, and TNF-α were measured in culture supernatants by ELISA. The frequency of PD-1+/CD4+ and Tim-3+/PD-1+/CD4+ T cells was significantly higher in patients with RA compared to that in controls (p = 0.0013 and p = 0.050, respectively). The percentage of Tim-3+/CD4+ T cells was similar in patients and controls (p = 0.4498). The RA patients have produced significant higher levels of TNF-α, IL-17, and IFN-γ than those of healthy controls (p = 0.0121, p = 0.0417, and p = 0.0478, respectively). Interestingly, an inverse correlation was found between the frequency of Tim-3+/CD4+ cells and DAS28 of RA patients (r = − 0.4696, p = 0.0493). Similarly, the percentage of Tim-3+/PD-1+/CD4+ T cells was also revealed an inverse correlation with DAS28 (r = − 0.5268, p = 0.0493). Moreover, significant positive correlations were detected between the concentrations of TNF-α (r = 0.6418, p = 0.0023) and IL-17 (r = 0.4683, p = 0.0373) with disease activity of RA patients. Our results indicate that Tim-3 and PD-1 are involved in immune dysregulation mechanisms of rheumatoid arthritis and could be considered as useful biomarkers for determination of disease activity and progression.
KeywordsDisease Activity Score PD-1 Rheumatoid arthritis Tim-3
The authors thank the patients and their families for their support, cooperation, and patience. We would like to thank the staff of the departments associated with the care and management of the patients. We also thank Mr. Ali Mosayebian for his collaboration in obtaining samples from normal volunteers.
The study was financially supported by Mazandaran University of Medical Sciences, grant number MCBRC-MAZUMS-1282.
Compliance with ethical standards
Informed consent was obtained from all participants, and the study was approved by Human Ethics Committee of Mazandaran University of Medical Sciences.
- 3.Broeren MG, de Vries M, Bennink MB, Arntz OJ, Blom AB, Koenders MI, van Lent PL, van der Kraan PM, van den Berg WB, van de Loo FA (2016) Disease-regulated gene therapy with anti-inflammatory interleukin-10 under the control of the CXCL10 promoter for the treatment of rheumatoid arthritis. Hum Gene Ther 27(3):244–254. https://doi.org/10.1089/hum.2015.127 CrossRefPubMedGoogle Scholar
- 5.Ceeraz S, Hall C, Choy EH, Spencer J, Corrigall VM (2013) Defective CD8+CD28+ regulatory T cell suppressor function in rheumatoid arthritis is restored by tumour necrosis factor inhibitor therapy. Clin Exp Immunol 174(1):18–26. https://doi.org/10.1111/cei.12161 CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Belkhir R, Burel SL, Dunogeant L, Marabelle A, Hollebecque A, Besse B, Leary A, Voisin AL, Pontoizeau C, Coutte L, Pertuiset E, Mouterde G, Fain O, Lambotte O, Mariette X (2017) Rheumatoid arthritis and polymyalgia rheumatica occurring after immune checkpoint inhibitor treatment. Ann Rheum Dis 76(10):1747–1750. https://doi.org/10.1136/annrheumdis-2017-211216 CrossRefPubMedGoogle Scholar
- 7.Bertsias GK, Nakou M, Choulaki C, Raptopoulou A, Papadimitraki E, Goulielmos G, Kritikos H, Sidiropoulos P, Tzardi M, Kardassis D, Mamalaki C, Boumpas DT (2009) Genetic, immunologic, and immunohistochemical analysis of the programmed death 1/programmed death ligand 1 pathway in human systemic lupus erythematosus. Arthritis Rheum 60(1):207–218. https://doi.org/10.1002/art.24227 CrossRefPubMedGoogle Scholar
- 14.Taghiloo S, Allahmoradi E, Tehrani M, Hossein-Nataj H, Shekarriz R, Janbabaei G, Abediankenari S, Asgarian-Omran H (2017) Frequency and functional characterization of exhausted CD8+ T-cells in chronic lymphocytic leukemia. Eur J Haematol 98:622–631. https://doi.org/10.1111/ejh.12880 CrossRefPubMedGoogle Scholar
- 15.Cai XZ, Huang WY, Qiao Y, Chen Y, Du SY, Chen D, Yu S, Liu N, Dou LY, Jiang Y (2015) Downregulation of TIM-3 mRNA expression in peripheral blood mononuclear cells from patients with systemic lupus erythematosus. Braz J Med Biol Res 48(1):77–82. https://doi.org/10.1590/1414-431x20143701 CrossRefPubMedGoogle Scholar
- 20.Wong M, La Cava A, Singh RP, Hahn BH (2010) Blockade of programmed death-1 in young (New Zealand black x New Zealand white)F1 mice promotes the activity of suppressive CD8+ T cells that protect from lupus-like disease. J Immunol 185(11):6563–6571. https://doi.org/10.4049/jimmunol.0903401 CrossRefPubMedGoogle Scholar
- 21.Raptopoulou AP, Bertsias G, Makrygiannakis D, Verginis P, Kritikos I, Tzardi M, Klareskog L, Catrina AI, Sidiropoulos P, Boumpas DT (2010) The programmed death 1/programmed death ligand 1 inhibitory pathway is up-regulated in rheumatoid synovium and regulates peripheral T cell responses in human and murine arthritis. Arthritis Rheum 62(7):1870–1880. https://doi.org/10.1002/art.27500 PubMedGoogle Scholar
- 26.Miko E, Meggyes M, Bogar B, Schmitz N, Barakonyi A, Varnagy A, Farkas B, Tamas P, Bodis J, Szekeres-Bartho J, Illes Z, Szereday L (2013) Involvement of Galectin-9/TIM-3 pathway in the systemic inflammatory response in early-onset preeclampsia. PLoS One 8(8):e71811. https://doi.org/10.1371/journal.pone.0071811 CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Hamel KM, Cao Y, Wang Y, Rodeghero R, Kobezda T, Chen L, Finnegan A (2010) B7-H1 expression on non-B and non-T cells promotes distinct effects on T- and B-cell responses in autoimmune arthritis. Eur J Immunol 40(11):3117–3127. https://doi.org/10.1002/eji.201040690 CrossRefPubMedPubMedCentralGoogle Scholar
- 28.Moriyama K, Kukita A, Li YJ, Uehara N, Zhang JQ, Takahashi I, Kukita T (2014) Regulation of osteoclastogenesis through Tim-3: possible involvement of the Tim-3/galectin-9 system in the modulation of inflammatory bone destruction. Lab Investig 94(11):1200–1211. https://doi.org/10.1038/labinvest.2014.107 CrossRefPubMedGoogle Scholar
- 32.Liu Y, Shu Q, Gao L, Hou N, Zhao D, Liu X, Zhang X, Xu L, Yue X, Zhu F, Guo C, Liang X, Ma C (2010) Increased Tim-3 expression on peripheral lymphocytes from patients with rheumatoid arthritis negatively correlates with disease activity. Clin Immunol 137(2):288–295. https://doi.org/10.1016/j.clim.2010.07.012 CrossRefPubMedGoogle Scholar
- 33.Reynolds J, Sando GS, Marsh OB, Salama AD, Evans DJ, Cook HT, Pusey CD (2012) Stimulation of the PD-1/PDL-1 T-cell co-inhibitory pathway is effective in treatment of experimental autoimmune glomerulonephritis. Nephrol Dial Transplant 27(4):1343–1350. https://doi.org/10.1093/ndt/gfr529 CrossRefPubMedGoogle Scholar
- 37.Bartosinska J, Zakrzewska E, Krol A, Raczkiewicz D, Purkot J, Majdan M, Krasowska D, Chodorowska G, Giannopoulos K (2017) Differential expression of programmed death 1 (PD-1) on CD4+ and CD8+ T cells in rheumatoid arthritis and psoriatic arthritis. Polish. Arch Intern Med 127(12):815–822. https://doi.org/10.20452/pamw.4137 Google Scholar
- 39.Lopa S, Leijs MJ, Moretti M, Lubberts E, van Osch GJ, Bastiaansen-Jenniskens YM (2015) Arthritic and non-arthritic synovial fluids modulate IL10 and IL1RA gene expression in differentially activated primary human monocytes. Osteoarthr Cartil 23(11):1853–1857. https://doi.org/10.1016/j.joca.2015.06.003 CrossRefPubMedGoogle Scholar