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食蟹猴腹主动脉补丁缝合术后的免疫排斥反应监测
引用本文:赵成江, 叶学军, 陈姣, 等. 食蟹猴腹主动脉补丁缝合术后的免疫排斥反应监测[J]. 器官移植, 2017, 8(2): 127-131. doi: 10.3969/j.issn.1674-7445.2017.02.007
作者姓名:赵成江  叶学军  陈姣  张汉成  周汇栋  邹志成  蔡志明  牟丽莎
作者单位:510275 广州,中山大学(赵成江、蔡志明、牟丽莎);深圳市第二人民医院(赵成江、叶学军、陈姣、张汉成、周汇栋、邹志成、蔡志明、牟丽莎)
基金项目:深圳市高水平医学学科建设专项基金2016031638 深圳市三名工程;深圳市科创委学科布局项目JCYJ20160229204849975 深圳市科创委企业工程中心项目GCZX2015043017281705 深圳市卫计委临床技术研究与转化类201501018
摘    要:目的  建立腹主动脉补丁缝合术的异种移植模型以及术后免疫排斥监测平台。方法  将野生型巴马猪的颈动脉,修剪成约2.5 cm×1.0 cm大小的梭形补丁,缝合于食蟹猴腹主动脉,未给予免疫抑制剂。观察受体猴一般情况。术后1年采用病理学检查观察移植物动脉补丁的形态学变化。分别在术前和术后7、14、28、49 d采集受体猴血液样本,利用猪的红细胞、外周血单核细胞(PBMC)检测受体猴血清中针对猪抗原的IgM和IgG抗体水平。采用血常规五分类法和流式细胞术检测受体猴血液中淋巴细胞计数。结果  3只移植猴存活状态良好。术后1年动脉补丁所在的血管壁外侧组织呈现深红色,苏木素-伊红(HE)染色显示有大量的红细胞和血小板沉积,其中有淋巴细胞的浸润。以猪的红细胞和PBMC为靶细胞,血清中的抗猪IgM和IgG抗体水平在术后28 d达到峰值,术后49 d时水平有所下降。淋巴细胞、T细胞亚群也是在术后28 d达到峰值,49 d开始回落。结论  动脉补丁缝合术是一种便捷可靠的异种移植模型。没有免疫抑制剂的条件下,受体维持了正常的生理状态。移植物可有效激活受体的免疫系统,诱导抗猪抗体的产生,引发细胞免疫排斥反应,因此该模型可用于监测异种移植过程中的免疫排斥反应。

关 键 词:异种移植   异种抗体   淋巴细胞   巴马猪   动物模型   流式细胞术   外周血单核细胞   红细胞
收稿时间:2017-01-03

Monitoring of immune rejection after abdominal aortic patch suture in cynomolgus monkeys
Zhao Chengjiang, Ye Xuejun, Chen Jiao, et al. Monitoring of immune rejection after abdominal aortic patch suture in cynomolgus monkeys[J]. ORGAN TRANSPLANTATION, 2017, 8(2): 127-131. doi: 10.3969/j.issn.1674-7445.2017.02.007
Authors:Zhao Chengjiang  Ye Xuejun  Chen Jiao  Zhang Hancheng  Zhou Huidong  Zou Zhicheng  Cai Zhiming  Mou Lisha
Affiliation:Sun Yat-sen University, Guangzhou 510275, China
Abstract:Objective To establish a platform to monitor the immune rejection after abdominal aortic patch suture in a xenotransplantation model. Methods The carotid was excised from wild-type Guangxi Bama pigs, cut into 2.5 cm × 1.0 cm pieces in shuttle shape and subsequently sutured to the abdominal aorta of cynomolgus monkeys. No immunosuppressive agent was administered. General conditions of the recipient monkeys were observed. The morphological changes of the graft artery were assessed by pathological examination at postoperative 1 year. Before and 7, 14, 28 and 49 d after surgery, the blood samples were collected from the recipient monkeys. The serum levels of IgM and IgG antibodies were quantitatively measured by the red blood cell and peripheral blood mononuclear cell (PBMC) from Guangxi Bama pigs. The quantity of lymphocytes in the recipient monkeys was detected by routine blood test and flow cytometry. Results All 3 monkeys undergoing transplantation survived well. At postoperative 1 year, the lateral tissues of the vascular wall at the artery graft were seen in dark red color. Hematoxylin-eosin (HE) staining revealed a large quantity of red blood cell and platelet deposition, accompanied with lymphocyte infiltration. Using porcine red blood cell and PBMC as target cells, the serum levels of anti-pig IgM and IgG antibodies peaked at postoperative 28 d, and slightly declined at postoperative 49 d. The quantity of lymphocytes and T cell subset also peaked at postoperative 28 d and began to decrease at postoperative 49 d. Conclusions Artery patch suture is a simple and reliable xenotransplantation model. The recipients can maintain normal physiological state without the use of immunosuppressive agents. The grafts can effectively activate the immune system of the recipients, induce the production of anti-pig antibodies and provoke cellular immune rejection. Therefore, this model can be utilized to monitor the immune rejection throughout the xenotransplantation process.
Keywords:Xenotransplantation  Xenoantibody  Lymphocyte  Guangxi Bama pig  Animal model  Flow cytometry  Peripheral blood mononuclear cell  Red blood cell
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