供体脾细胞输注诱导大鼠异体肢体移植耐受的实验研究

发布时间：2019-07-06 09:14

【摘要】： 目的探讨联合应用供体特异性脾细胞输注(donor specificspleen cell tranfusion,DST)和环磷酰胺诱导移植耐受的作用及其机制。方法选用20只雄性Wistar大鼠和20只雌性SD大鼠分别作为肢体移植的供体和受体,2只DA大鼠作为无关第三品系用以检测移植耐受状态的特异性。将20只受体大鼠随机分为4组,即无处理对照组、单纯SC组、单纯CP组和实验组(SC+CP),每组5只。诱导耐受的方案是在肢体移植手术当天经尾静脉给受体SD大鼠输注供体Wistar大鼠的脾细胞(3×10~8),术后第2天给予受者大鼠腹腔注射环磷酰胺150mg/Kg。观察移植肢体存活时间、对受体SD大鼠作单向混合淋巴细胞反应,并用多聚酶链式反应(PCR)检测嵌合体的形成。结果实验组肢体移植物存活时间(18.4±1.82d)比无处理对照组(8.4±1.14d)、单纯SC组(9.0±1.23d)、单纯CP组(9.6±1.14d)明显延长(P＜0.01)。混合淋巴细胞反应实验组特异性抑制率74.2±4.92%明显高于单纯SC组37.2±3.90%和单纯CP组39.4±4.16%(P＜0.01)。2-4组微嵌合呈阳性。结论1 Wistar大鼠—SD大鼠进行后肢肢体移植后排斥反应剧烈且恒定,移植肢体便于观察,大鼠外周血可进行各种血清学检测。因此,此移植模型是肢体移植的理想动物模型。2供体特异性脾细胞输注联合环磷酰胺腹腔注射可诱导受体鼠的免疫耐受。3本实验所诱导的免疫耐受是不完全和暂时的,对供体具有特异性。4微嵌合体的形成和嵌合水平的高低可能与免疫耐受的产生及维持有关。
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图片说明：各组间SRY与p.ac舫n灰度比位图
[Abstract]:Objective to investigate the effect and mechanism of donor specific spleen cell infusion (donor specificspleen cell tranfusion,DST) and cyclophosphamide (cyclophosphamide) on transplantation tolerance. Methods 20 male Wistar rats and 20 female SD rats were used as donors and recipients of limb transplantation, and 2 DA rats were used as unrelated third strains to detect the specificity of transplantation tolerance. Twenty recipient rats were randomly divided into 4 groups: control group without treatment, simple SC group, simple CP group and experimental group with 5 rats in each group. The regimen of inducing tolerance was to infuse the spleen cells (3 脳 10 ~ 8) of recipient SD rats via tail vein on the day of limb transplantation, and to inject cyclophosphamide 150 mg 路kg ~ (- 1) intraperitoneally on the second day after operation. The survival time of the recipient SD rats was observed, and the chimerism was detected by polymerase chain reaction (PCR). Results the survival time of limb graft in the experimental group (18.4 卤1.82 days) was significantly longer than that in the control group (8.4 卤1.14 days), SC group (9.0 卤1.23 days) and CP group (9.6 卤1.14 days). The specific inhibition rate of mixed lymphocytes reaction group was 74.2 卤4.92%, which was significantly higher than that of simple SC group (37.2 卤3.90%) and CP group (39.4 卤4.16%). Conclusion 1 the rejection of hindlimb limb transplantation in Wistar rats and SD rats is severe and constant, and the limb transplantation is easy to observe, and various serological tests can be carried out in peripheral blood of rats. Therefore, this transplantation model is an ideal animal model for limb transplantation. 2 donor-specific spleen cell infusion combined with cyclophosphamide intraabdominal injection can induce immune tolerance in recipient mice. 3 the immune tolerance induced by this experiment is incomplete and temporary, and has specificity to donors. 4 the formation and chimerism of microchimerism may be related to the generation and maintenance of immune tolerance.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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