Clinical Rheumatology

, Volume 37, Issue 8, pp 2063–2071 | Cite as

Analysis of PD-1 and Tim-3 expression on CD4+ T cells of patients with rheumatoid arthritis; negative association with DAS28

  • Zohreh Koohini
  • Hadi Hossein-Nataj
  • Maryam Mobini
  • Aref Hosseinian-Amiri
  • Alireza Rafiei
  • Hossein Asgarian-OmranEmail author
Original Article


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.


Disease 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.

Funding information

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.




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Copyright information

© International League of Associations for Rheumatology (ILAR) 2018

Authors and Affiliations

  1. 1.Department of Immunology, School of MedicineMazandaran University of Medical SciencesSariIran
  2. 2.Department of Rheumatology, Imam Khomeini Hospital, School of MedicineMazandaran University of Medical SciencesSariIran
  3. 3.Immunogenetics Research Center, School of MedicineMazandaran University of Medical SciencesSariIran

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