| Background Interleukin-10 was originally characterized as a T-cell growth and differentiation factor . Because of its ability to reduce the production of IL-2, IFN, and TNF- by murine Thl cells, it was named CK synthesis inhibitory factor. In human beings, IL-10 is produced by a variety of cell types, including T lymphocytes (particularly the Th2 type), B lymphocytes, macrophages, and NK cells. Human IL-10 inhibits the production of IL-2, IFN, and granulocyte-macrophage colony-stimulating factor (GM-CSF) as well as the proliferative response of Thl cells. Interleukin-10 can impair the antigen-presenting properties of DCs by reducing the expression of antigen-presenting and costimulatory molecules and by interfering with the maturation of monocytes to DCs As a result of these biologic properties, IL-10 is classified as a Th2 type CK with potent anti-inflammatory and immunosuppressive activities. Accordingly, its use has been advocated and is currently under clinical evaluation in human beings for the therapyof autoimmune diseases and to induce tolerance after hepatic transplantation. An active transposase called Sleeping Beauty(SB) was reconstructed recently, which provided somatic intergration and long-term transgene expression. Objective To clone human IL-10 gene and construct Sleeping Beauty transposon system. Methods cDNA fragment encoding hIL-10 gene was amplified from human peripheral normal white blood cells by RT-PCR. After confirmed hIL-l0DNA sequencing, IL-10 cDNA was inserted into pcDNA3 vector, this recombinant expressing vector was used to transfect CHO cell. The IL-10 molecules expressed were detected by ELISA and the role of inhibition of IL-10 was determined with MTT. Results The hIL-10 was expressed in the CHO and it blocked the lymphocyte transformation. Conclusion The cloning of human IL-10 gene and constructing Sleeping Beauty transposon system provide the foundation for further investigate the role of hIL-10 in transplantation immunity. Keywords human interleukin-10 clonePartThe study of human interleukinlO transferred with Sleeping Beautytransposon system prevents liver transplant rejection in rats Background Immunosuppressive regimens used to prevent rejection after transplantation are associated with increased risks of opportunisticinfections, malignancy, metabolic complications, and heart and vascular diseases. Despite immunosuppressive therapy, acute allograft rejection following liver transplantation occurs in 40% to 70% of patients. Cytokines are involved in rejection of human liver grafts . Thltype cytokines including IL-2, TNF, and IFN-y, mediate cellular immune responses, while Thtype cytokines like 1L-4 and L0 have been shown to inhibit the development and function of Thl-cells, suppress inflammation. Th 1-cytokines are mainly involved in allograft rejection, by upregulation of MHC class I and II expression, stimulating macrophage function (TNF-a and IFN-y), by endothelial cell activation, up-regulation of cell adhesion molecules, and facilitating the recruitment and activation of leukocytes (TNF-a). Plasma levels of TNF-a are raised during acute rejection episodes following heart, kidney, and liver transplantation. Evidence from animal transplant models has suggested a role for Th2-cytokines like IL-4 and IL-10 in promoting graft survival. Their mechanism of action would be in down-regulating pro-inflammatory cytokine production. IL-10 may also down-regulate T-cell activation by decreasing MHC class and B7 expression on antigen-presenting cells. In this study, donor liver were transferred hIL-10 with with Sleeping Beauty transposon system via portal vein in cold preservation, we studied the effect on long-term graft survival and the mechanism of maintenance of tolerance in rat allogeneic liver12transplantation and compared with immunosuppressive drug FK506.Objective To study the effect of interleukinl0 transferred with Sleeping Beauty transposon system on long-term graft survival and the mechanism of maintenance of tolerance in rat allogeneic liver transplantation...
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