Methrotexate Treatment Inmunomodulates Abnormal Cytokine Expression by T CD4 Lymphocytes Present in DMARD-Naïve Rheumatoid Arthritis Patients

Abstract

CD4⁺T-lymphocytes are relevant in the pathogenesis of rheumatoid arthritis (RA), however, their potential involvement in early RA remains elusive. Methotrexate (MTX) is a commonly used disease-modifying antirheumatic drug (DMARD), but its mechanism has not been fully established. In 47 new-onset DMARD-naïve RA patients, we investigated the pattern of IFNγ, IL-4 and IL-17A expression by naïve (T_N), central (T_CM), effector memory (T_EM) and effector (T_E) CD4⁺ subsets; their STAT-1, STAT-6 and STAT-3 transcription factors phosphorylation, and the circulating levels of IFNγ, IL-4 and IL-17. We also studied the RA patients after 3 and 6 months of MTX treatment and according their clinical response. CD4⁺T-lymphocyte subsets and cytokine expression were measured using flow cytometry. New-onset DMARD-naïve RA patients showed a significant expansion of IL-17A⁺, IFNγ⁺ and IL-17A⁺IFNγ⁺ CD4⁺T-lymphocyte subsets and increased intracellular STAT-1 and STAT-3 phosphorylation. Under basal conditions, nonresponder patients showed increased numbers of circulating IL-17A producing T_N and T_MC CD4⁺T-lymphocytes and IFNγ producing T_N, T_CM, T_EM CD4⁺T-lymphocytes with respect to responders. After 6 months, the numbers of CD4⁺IL-17A⁺T_N remained significantly increased in nonresponders. In conclusion, CD4⁺T-lymphocytes in new-onset DMARD-naïve RA patients show IL-17A and IFNγ abnormalities in T_N, indicating their relevant role in early disease pathogenesis. Different patterns of CD4⁺ modulation are identified in MTX responders and nonresponders.

Keywords: CD4+ T-lymphocytes; IFNγ, IL-17A; STAT expression; methotrexate response; naïve rheumatoid arthritis.