In vitro generation of IL-35-expressing human Wharton's Jelly-derived Mesenchymal Stem Cells using lentiviral vector

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. copyright© summer 2015,iran j allergy asthma immunol. all rights reserved.