Objectives: Balneotherapy with mud application (mud therapy) is a common hydrothermal intervention for the treatment and rehabilitation of elderly patients with osteoarthritis, leading to anti-inflammatory effects. The main purpose of this investigation was to study a role for regulatory T cells in these anti-inflammatory effects. The second objective was to assess whether the neutrophil-mediated innate response is affected by these anti-inflammatory effects. Methods: Thirty-six elderly patients with knee osteoarthritis underwent a 10-day cycle of balneotherapy at a spa center. They received daily sessions of whole-body mud therapy at 40-42 C, using mineral-medicinal water and mud. IL-8 and TGF-b serum concentrations, percentage of circulating CD4 þ CD25 þ FOXP3 þ and CD8 þ CD28regulatory T cells, and neutrophil phagocytic capacity were evaluated at baseline and at the end of the intervention. Clinical assessments included knee flexion and extension angle, pain, stiffness, physical function and health-related quality of life. Results: All clinical outcomes significantly improved. Circulating concentrations of IL-8 and TGF-b decreased, which correlated with decreased pain and improved knee flexion, respectively. Percentage of CD4 þ regulatory T cells decreased, whereas CD8 þ regulatory T cells increased. Neutrophil functional capacity increased. Conclusions: Balneotherapy with mud application was effective in the management of osteoarthritis symptoms. The anti-inflammatory effect mediated by cytokines contributed to the improvement in pain and joint function; and changes in the circulating percentage of regulatory T cells seem to be involved in the anti-inflammatory effects. Improvement in neutrophil function after mud therapy reflects an optimal bioregulatory effect on the inflammatory and innate responses.