转基因CTLA4-Ig诱导大鼠肝移植免疫耐受的实验研究

发布时间：2018-03-15 09:37

本文选题：CTLA4-Ig　切入点：肝移植　出处：《中国人民解放军军事医学科学院》2007年硕士论文　论文类型：学位论文

【摘要】： 目的：通过建立大鼠原位肝移植模型，探讨CTLA4-Ig抗排斥反应和诱导免疫耐受的作用。方法：采用近交系雌性DA大鼠作供体、雌性LEW大鼠作受体，“二袖套法”建立肝脏移植模型20例。术前随机分为2组：A组即排斥组(DA→LEW，n=10)，只进行原位肝移植；B组即CTLA4-Ig基因治疗组(DA→LEW，n=10)，在供肝冷保存时，将CTLA4-Ig基因导入大鼠移植肝。A、B两组受体分别于术后3、7、16d经眼球取血获得血清标本。术后7d各组随即处死2例，收集肝脏标本。血清标本检测IL-2、ALT、AST、TBIL的浓度。肝脏标本观察移植物病理学变化。结果：成功建立了大鼠原位肝移植模型。DA→LEW近交系大鼠肝移植后移植排斥反应强烈，在不使用免疫抑制剂的情况下，生存时间＜14天，是进行肝移植免疫研究的可靠模型。术前大鼠血清IL-2浓度362.31±49.26 ng／L。肝移植术后，IL-2的浓度在A组中升高到较高水平，而在B组中则维持一个较低的水平。术后3、7d，A组与B组血清IL-2的浓度存在非常显著的差异。术前、术后3d，A组与B组ALT、AST、TBIL之间无统计学差异；术后7d，A组浓度明显高于B组(P＜0.05)。A组与B组，在术后7d行移植肝取材，病理学证实：A组移植肝发生严重的细胞性排斥，B组可见移植肝组织损伤程度轻。结论：在DA→LEW大鼠原位肝移植的基础上，，于供肝冷保存时应用腺病毒载体介导CTLA4-Ig基因转染供肝，可以抑制同种异体免疫反应，在不使用常规免疫抑制剂的情况下，能延长移植肝的存活时间，降低受体对供体移植物的免疫反应性。
[Abstract]:Aim: to establish an orthotopic liver transplantation (OLT) model in rats to investigate the effects of CTLA4-Ig on anti rejection and immune tolerance. Methods: twenty cases of liver transplantation model were established by using inbred female DA rats as donor and female LEW rats as recipients. Before operation, 20 cases of liver transplantation were randomly divided into two groups: group A: rejection group. 鈫扡EWN 10, orthotopic liver transplantation group B (CTLA4-Ig gene therapy group). 鈫扺hen the donor liver was cryopreserved, the CTLA4-Ig gene was introduced into the transplanted liver of rats. The blood samples were collected from the eyeball on the 7th and 16th day after operation, and 2 patients in each group were killed 7 days after operation. Liver samples were collected. Serum samples were collected to detect the concentration of IL-2ALT TBIL. Liver specimens were observed for the histopathological changes of grafts. Results: the rat orthotopic liver transplantation model was successfully established. DA. 鈫扵he rejection of LEW inbred rats after liver transplantation was strong, and the survival time was less than 14 days without the use of immunosuppressants. The serum IL-2 concentration of rats before operation was 362.31 卤49.26 ng / L. The level of IL-2 increased to a higher level in group A after liver transplantation. There was significant difference in serum IL-2 concentration between group A and group B. there was no significant difference between group A and group B 3 days after operation, and the concentration of serum IL-2 in group A was significantly higher than that in group B at 7 days after operation (P < 0.05), but there was no significant difference between group B and group A on the 3rd day after operation (P < 0.05), but there was no significant difference between group A and group B on the 7th day after operation (P < 0.05), but there was no significant difference between group B and group A on the 3rd day after operation (P < 0.05). The grafts were taken on the 7th day after operation. The pathological evidence showed that the grafts in group A had severe cellular rejection and that the grafts in group B had slight liver tissue damage. Conclusion: in DA. 鈫扥n the basis of orthotopic liver transplantation (OLT) of LEW rats, CTLA4-Ig gene transfection mediated by adenovirus vector during cold preservation of donor liver could inhibit the allogeneic immune response without routine immunosuppressive agents. It can prolong the survival time of the transplanted liver and decrease the immunoreactivity of the recipient to the donor graft.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R657.3;R392

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