Objective: The purpose of this study was to investigate the therapeutic effect of hydroxychloroquine on rheumatoid arthritis (RA) and to analyze the treatment's effect on patients' Treg, Th1, and Th2 cells. Methods: A total of 160 RA patients admitted to our hospital were divided into a study group (SG, n = 80) or a control group (CG, n = 80) according to the treatment each patient received. The CG was treated with methotrexate, and the SG was treated with methotrexate and hydroxychloroquine. The treatment efficiency, the biochemical index levels, the CD4+ T lymphocyte expressions, the differences in their joint symptoms before and after the treatment, and the differences in the patients' pain levels before and after the treatment were compared between the two groups. The incidence of adverse reactions was also compared. Results: The effective rate of treatment in the SG was 97.50%, which was higher than the rate of 82.50% in the CG (P < 0.05). After 6 months of treatment, the incidence of adverse reactions in the SG was significantly higher than it was in the CG. CRP, ESR, and RF in the SG were all lower than they were in the CG (P < 0.05). The IFN-gamma, IL-10, and IL-35 expression levels in the SG were significantly lower than they were in the CG after the treatment (P < 0.05). The tender joint counts (TJC) and swollen joint counts (SJC) showed no significant differences between the two groups before the treatment (P > 0.05) and were lower in the SG than they were in the CG (P < 0.05). There were no significant differences in the VAS, PGA, or PhGA between the two groups before the treatment, and the above indicators in the SG were all lower than they were in the CG after the treatment (P > 0.05). Conclusion: Hydroxychloroquine therapy has a good clinical effect on RA patients, for it can significantly improve the expression abnormalities of CD4+ T lymphocytes, improve patients' laboratory parameters and clinical symptoms, and achieve high treatment safety.