Tehran University of Medical Sciences Iranian Journal of Allergy, Asthma and Immunology 1735-1502 14 4 2015 10 20 416 426 EN Afshin Amari Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Massoumeh Ebtekar Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Seyed Mohammad Moazzeni Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Masoud Soleimani Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran AND Department of Stem Cells Biology, Stem Cell Technology Research Center, Tehran, Iran. Leila Mohammadi-Amirabad Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Mohammad Taher Tahoori Department of Immunology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. Mohammad Massumi Department of Stem Cells Biology, Stem Cell Technology Research Center, Tehran, Iran AND National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. 2015 10 19 2015 10 19 Human Wharton’s Jelly-derived Mesenchymal Stem Cells (hWJ-MSCs) are easily available cells without transplant rejection problems or ethical concerns compared to bone-marrow-derived MSCs for prospective clinical applications. These cells display immunosuppressive properties and may be able to play an important role in autoimmune disorders. Regulatory T-cells (Treg) are important to prevent autoimmune disease development. Interleukin 35 (IL-35) induces the proliferation of Treg cell populations and reduces the activity of T helper 17 (Th17) and T helper 1 (Th1) cells, which play a central role in initiation of inflammation and autoimmune disease.Recent studies identified IL-35 as a new inhibitory cytokine required for the suppressive function of Treg cells. We created IL-35-producing hWJ-MSCs as a good vehicle for reduction of inflammation and   autoimmune   diseases.   We   isolated   hWJ-MSCs   based   on   explant   culture.   HWJ-MSCs were transduced at MOI=50 (Multiplicity of Infection) with lentiviral particles harboring murine Interleukin 35 (mIL-35). Expression of IL-35 in hWJ-MSCs was quantified by an IL-35 ELISA kit.IL-35 bioactivity was analyzed by inhibiting the proliferation of mouse splenocytes using CFSE cell proliferation kit. Frequency of CD4+CD25+CD127low/neg Foxp3+ Treg cells was measured by flow cytometry. There was an up to 85% GFP positive transduction rate, and the cells successfully released a high level of mIL-35 protein (750 ng/ml). IL-35 managed to inhibit CD4+ T cell proliferation with PHA, and improved the frequency of Treg cells.Our data suggest that transduced hWJ-MSCs overexpressing IL-35 may provide a useful approach for basic research on gene therapy for autoimmune disorders. https://ijaai.tums.ac.ir/index.php/ijaai/article/view/589 https://ijaai.tums.ac.ir/index.php/ijaai/article/download/589/524