论文部分内容阅读
背景:前期的研究表明,通过受体全身注射T细胞活化所需的B7/CD28共刺激信号阻断剂细胞毒性T淋巴细胞相关抗原4Ig可以明显延长大鼠移植肾存活,但所需剂量大、影响范围广、特异性差,可能造成全身性不良反应。目的:为了提高细胞毒性T淋巴细胞相关抗原4Ig治疗的靶向性,采用细胞毒性T淋巴细胞相关抗原4Ig基因对肾移植供、受体树突状细胞进行体外修饰,观察两种树突状细胞单独以及联合应用对移植肾存活的影响。设计、时间及地点:随机对照动物实验,于2003-04/2004-07在解放军全军肾病中心实验室完成。材料:肾移植供体为近交系Brown-Norway(BN)大鼠,受体为近交系Lewis大鼠,Wistar大鼠作为无关淋巴细胞供体。方法:分离培养供、受体大鼠骨髓源性树突状细胞,以细胞毒性T淋巴细胞相关抗原4Ig基因重组腺病毒转染供、受体大鼠骨髓源性树突状细胞。构建大鼠肾移植模型,将单独的供、受体树突状细胞,混合的供、受体树突状细胞于肾移植前24h经股静脉注入受体,以未注射树突状细胞的受体为对照。主要观察指标:①移植肾存活时间。②术后20d应用MTT法检测受体脾细胞对供体及无关抗原刺激反应程度。结果:与对照组比较,受体树突状细胞未能延长移植肾存活,但与供体树突状细胞联合应用时可使移植肾存活时间较单独应用供体树突状细胞时明显延长(P<0.01)。注入供体树突状细胞与混合的供、受体树突状细胞大鼠的脾细胞对供体抗原刺激的反应均明显低于对照组(P<0.01),而对无关抗原刺激的反应与对照组无差异。结论:细胞毒性T淋巴细胞相关抗原4Ig基因修饰的供体树突状细胞可以明显延长大鼠移植肾存活时间,受体树突状细胞无此作用,但两种树突状细胞联合应用可以使移植肾获得更长的存活时间。
BACKGROUND: Previous studies have shown that 4Ig, a B7 / CD28 costimulatory signal blocker required for systemic activation of T cells, can significantly prolong the survival of rat renal allografts, but require large doses, A wide range of effects, poor specificity, may cause systemic adverse reactions. OBJECTIVE: To improve the targeting of cytotoxic T lymphocyte-associated antigen 4Ig, dendritic cells donated and recipient dendritic cells were treated with cytotoxic T lymphocyte-associated antigen 4Ig gene in vitro. Two kinds of dendritic cells The effects of single and combined application on graft survival. DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed in the People's Liberation Army's Nephrology Center Laboratory from April 2003 to July 2004. MATERIALS: The renal transplant recipients were Brown-Norway (BN) rats inbred line. The recipients were inbred Lewis rats and Wistar rats served as unrelated lymphocyte donors. Methods: Bone marrow-derived dendritic cells from donor and recipient rats were isolated and cultured. Bone marrow-derived dendritic cells from donor and recipient rats were transfected with 4Ig recombinant adenovirus containing cytotoxic T lymphocyte antigen. The rat renal transplantation model was established, dendritic cells for donor and recipient, mixed donor and recipient dendritic cells were injected into the recipient via the femoral vein 24h before renal transplantation. The dendritic cells Body as a control. MAIN OUTCOME MEASURES: ① graft survival time. ②Twenty days after operation, the response of donor splenocytes to donor and unrelated antigen was detected by MTT assay. Results: Compared with the control group, the recipient dendritic cells failed to prolong the survival of the grafted kidneys. However, when compared with the donor dendritic cells, the survival time of the grafted kidneys was significantly longer than that of the donor dendritic cells alone P <0.01). The response of splenocytes injected into donor dendritic cells and mixed donor and recipient dendritic cells to donor antigen was significantly lower than that of control (P <0.01), while the response to unrelated antigen stimulation No difference in the control group. CONCLUSION: Donor dendritic cells modified with 4Ig gene of cytotoxic T lymphocyte antigen can prolong the survival time of rat renal allografts without receptor dendritic cells. However, the combination of two dendritic cells can make Graft kidney to obtain longer survival time.