Treatment Of Ulcerative Colitis By Interleukin-35Modified Mesenchymal Stem Cells In Mice

This study was designed to investigate interleukin-35(IL-35) modified mesenchymal stem cells as treatment of ulcerative colitis in mice; After application of genetic engineering techniques to express the IL-35gene was cloned into a lentiviral vector infection to mesenchymal stem cells, We used ELISA method to detect gene-modified mesenchymal stem cells the expression of IL-35protein; We established a stable model of ulcerative colitis in mice by intravenous injection of IL-35gene-modified mesenchymal stem cells, to observe its treatment of acute ulcerative colitis in mice. We found IL-35gene modified MSCs could be a novel and promising approach in the treatment of UC.Partâ…  Preparation of IL-35modified mesenchymal stem cells of C57BL/6mice.Object:Construction of IL-35overexpression lentivirus vector, with its infection to mesenchymal stem cells (MSCs) to preparation of IL-35modified MSCs. Methods: Adipose-derived MSCs was obtained using type I collagenase from mouse fat, and characterized using flow cytometry (FCM) and differentiate to adipocytes and osteogenic progenitors. IL-35gene was integrated to working plasmid of lentivirus, with its co-transfected to packaging cells with packaging plasmids, IL-35overexpression lentivirus vector (IL-35-LV) was made. After infection of IL-35-LV to MSCs, enzyme-linked immunosorbent assay (ELISA) was used to assay IL-35expression in the cell supernatant. Result:Isolation of adipose-derived MSCs was success; After infection of IL-35-LV, MSCs can secrete IL-35.Partâ…¡ Construction of acute ulcerative colitis model in C57BL/6miceObject:To construct acute ulcerative colitis model of C57BL/6mice. Methods:Fed mice with different concentration of dextran sulfate (DSS) for7days. Evaluation of the ulcerative colitis (UC) was made by determination of weight change and pathology. Result:2%DSS can induce UC of C57BL/6mice successfully. Partâ…¢ Study of IL-35modified MSCs protect against ulcerative colitis in C57BL/6miceObject:To investigate the effects of IL-35modified MSCs on inflammation development and clinical outcomes in mice model of UC. Methods:The mice were administrated with IL-35gene modified MSCs, wild type MSCs or control saline solution intravenously on the2nd,4th and6th day of DSS feeding. The effects of the treatment were evaluated by determination of weight change, pathology, cytokine expression, lymphocyte subsets variation. Result:Compare to wild type MSCs and saline control, administration of IL-35gene modified MSCs alleviated UC development in mice fed with DSS. Tregs in LPL of IL-35gene modified MSCs treated mice is higher than the other two groups. Administration of IL-35gene modified MSCs upregulated IL-17expression and downregulated TNF-a and IFN-y expression in mice colon tissue.Conclusion:IL-35gene modifed MSCs may protect against DSS induced UC by up regulated the expression of IL-17, while IL-17has protect effect on DSS induced UC. Down regulation of TNF-a and IFN-y plays an important role in regulation of intestinal immune responses by controlling overwhelming T-cell activation. IL-35gene modified MSCs could be a novel and promising approach in the treatment of UC.