Secretory factors produced by adipose mesenchymal stem cells downregulate Th17 and increase Treg cells in peripheral blood mononuclear cells from rheumatoid arthritis patients

We aimed to assess the immunoregulatory effects of secretory factors produced by adipose tissue-derived MSC (AT-MSC) on Th17 and Treg subsets from patients with rheumatoid arthritis (RA). 17 patients with active disease matching the ACR/EULAR 2010 criteria for RA were included. Patients' peripheral blood mononuclear cells (PBMC) were cultured in AT-MSC-conditioned medium (AT-MSCcm) and in control medium. The cytokine production of AT-MSC and PBMC was quantified by ELISA. Th17 and Treg were determined by flow cytometry. AT-MSCcm contained: IL-6, IL-17, IL-21, CCL2, CCL5, IL-8, sVEGF-A and PGE(2). Cultivation of patients' PBMC with AT-MSCcm increased TGF-1 (8318pg/ml; IQR 6327-11,686) vs control medium [6227pg/ml (IQR 1681-10,148, p=0.013)]. PBMC cultivated with AT-MSCcm downregulated TNF-, IL-17A, and IL-21 compared to control PBMC: 5pg/ml IQR (1.75-11.65) vs 1pg/ml (IQR 0.7-1.9), p=0.001; 4.2pg/ml (IQR 3.1-6.1) vs 2.3pg/ml (IQR.75-5.42), p=0.017; 66.9pg/ml (IQR 40.6-107.2) vs 53pg/ml (IQR 22-73), p=0.022. Th17 decreased under the influence of AT-MSCcm: 10.13 +/- 3.88% vs 8.98 +/- 3.58%, p=0.02. CD4(+)FoxP3(+), CD4(+)CD25(-)FoxP3(+), and CD4(+)CD25(+)FoxP3(+) was 11.35 +/- 4.1%; 7.13 +/- 3.12% and 4.22 +/- 2% in control PBMC. Accordingly, CD4(+)FoxP3(+), CD4(+)CD25(-)FoxP3(+), and CD4(+)CD25(+)FoxP3(+) significantly increased in PBMC cultured with AT-MSCcm: 15.6 +/- 6.1%, p=0.001; 9.56 +/- 5.4%, p=0.004 and 6.04 +/- 3.6%, p=0.001. All these effects could define MSC-based approaches as adequate avenues for further treatment development in RA.