Infusion AdCTLA4-Ig Transfection Of Donor-specific Bone Marrow Mesenchvmal Stem Cells In Rat Limb Allotransplantation Specific Immune Tolerance

Objective To investigate the effect and mechanisms of transplantation induced by infusion CTLA4Ig-BMSCs via the tail vein.To provide an new protocol and idea for inducing limb transplantation toleranceMethods1、With AdEasy vector system, the recombinant adenoviruses containing CTLA4—Ig gene was constructed and the viral titer was measured.BMSCs were infected with the recombinant adenoviruses and the expression of CTLA4-Ig was detected by RT--PCR.2. Build limb transplantation in rats were randomly divided into A, B, C, D, four groups.Group A:blank control group Group B:infusion AdCTLA4Ig via the tail vein.Group C:infusion BMSCs via the tail vein Group D:infusion CTLA4Ig-BMSCs via the tail vein。The survival of grafts and transplantation rats were observed after transplantation. The morphological changes of grafts were observed by hematoxylin. eosin staining; cytokines were quantified by ELISA.Results (1)The recombinant adenovirus containing CTLA4-Ig gene was constmcted successfully and its titer was1.0*109efu/ml. The recombinant virus AdCTLA4Ig prepared in this study efficiently infected BMSCs and the infected cells expressed recombinant protein CTLA4Ig。(2) The survival of grafts in group D was significant longer than that in group A, B and D respectively(p<0.05); The survival of group B and C were significantly prolonged compared with group A(p<0.05)and the survival of group B was not significantly different with group C(p>0.05).(3) On day7post—transplantation, hematoxylin—eosin staining of grafts. Blank control group shows severe infiltration of lymphocytes; group B and C shows moderate infiltration of lymphocytes; groupD shows mild infiltration of lymphocytes。Conclusion (1) Recombinant adenoviruses could be obtained quickly and simply by using AdEasy system。(2) Via the tail vein infusion CTLA4Ig-BMSCs can induce immune tolerance, and superior to the single infusion AdCTLA4-Ig and BMSCs group.