Experimental Study On The Effect Of Local CTLA4Ig Gene Transfection On The Acute Rejection Of Small Bowel Allografts In Rats

Development of Segmental Heterotopic Small Bowel Transplantation Model in RatsObjective: To establish small bowel transplantation (SBT) model in rats in order to investigate the rejection of SBT. Method : Segmentalheterotopic SBT was performed on adult rats. A combination of aorta toinfrarenal aorta anastomosis and portal vein to infrarenal vein cavaanastomosis was adopted for small bowel graft revascularizing. Result: In the % cases receiving SBT, the 5-days survival rate of 56 cases undergoing SBT after the model stabilized was 85.71%. Conclusions: The successful rate of this model development is satisfactory, and the model is appropriate for rejection study of SBT.The Possibility of Local Transfection of CTLA4Ig Gene intoSmall Bowel Allografts in RatsObjective: To evaluate the possibility of local expression of CTLA4Iggene in small bowel allografts.Methods: The donor small bowels were perfused ex vivo with thereconstructed plasrnid of CTLA41g cDNA packaged with lipofectinvector via intra- superior mesenteric artery before transplantation. TheCTLA4Ig trans gene expression in the small bowel allografts was assessedby inimunohistoiogy and reverse inmscriptase polymerase chain reaction(RT-PCR) after transplantation.Results: {mmunohistoiogy and RT-PCR demonstrated abundantexpression of CTLA4Ig transgene in the allografts 48 hours aftertransplantation.Conclusion: A single ex vivo intra-superior mesenteric artery infusion ofreconstructed plasmid of CTLA4Ig cDNA packaged with lipofectin caninduce efficient transduction of the small bowel allografts.Effect of local CTLA4Ig gene transfection on the acute rejection of small bowel allografts in ratsObjective: To evaluate the local expression of CTLA4Ig gene in small bowels and its action on preventing acute rejection of the small bowel allografts.Methods: Wistar rats underwent heterotopic small bowel transplantation from SD rats. The recipients were divided into experimental group (allografts were transfected with CTLA4Ig gene) and control group (non CTLA4Ig gene transfected) randomly. In the experimental group, the donor small bowels were perfused ex vivo with me reconstructed plasmid of CTLA4Ig cDNA packaged with lipofectin vector via intra-superior mesenteric artery before transplantation, and the CTLA41g expression in the small bowel grafts post-transplantation was assessed by immunohistology. On day 3, 7 and 10 post-transplantation, the allografts in each group were harvested for the examination of histology and assayof apoptosis.Results: Small bowel allografts treated with CTLA4Ig cDNA showed abundant CTLA4Ig expression after transplantation. Acute rejection grade I on day 7 and grade on day 10 after transplantation was noticed in the control allografts, and a dramatically increase number of apoptotic enterocytes in parallel to the progressive rejection could be recognized. In contrast, the allografts treated with CTLA4Ig cDNA showed nonspecific histological changes and only a few apoptotic enterocytes was found after transplantation.Conclusion: Local CTLA4Ig gene transfection of small bowel allograft is feasible, and the local CTLA4Ig expression in the allograft can prevent acute rejection after transplantation.Effect of regional CTLA4Ig gene transfection on the rejectiveresponsiveness of small bowel recipients in ratsObjective: To evaluate the local expression of CTLA4Ig gene in smallbowels and its action on the rejective responsiveness of small bowelrecipients in rats.Methods: The donor small bowels were perfused ex vivo with thereconstructed plasmid CTLA4Ig cDNA packaged with lipofectin vectorvia intra- superior mesenteric artery before transplantation. The CTLA4Igtransgene expression in the small bowel allografts was assessed byimmunohistology and reverse transcriptase polymerase chain reaction,and the survival time of the aliografts was observed after transplantation.Results: Immunohistology and RT-PCR demonstrated expression of CTLA4i...