肾移植术后抗体介导排斥反应模型的构建与鉴定
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国家自然科学基金青年基金(81870512,81570676,81770751,81100532,81470981);江苏省“六大人才高峰”资助(2010WSN?56,2011?WS?033);江苏省“科教兴卫”工程医学重点人才培养资助(RC2011055);江苏省卫生厅面上项目(H2009907);江苏省“333高层次人才培养工程”(2011,2013)


Construction and identification of an antibody⁃mediated rejection model after kidney transplantation
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    摘要:

    目的:构建并鉴定一种肾移植术后抗体介导排斥反应(antibody-mediated rejection,AMR)的动物模型。方法:AMR动物模型采用Wistar大鼠作为供体,SD大鼠作为受体。将Wistar大鼠背部皮肤移植至SD大鼠背部,构建同种异体大鼠皮肤移植模型,分别在皮肤移植后1、3、7、10、14、21、28、35 d取大鼠血清,采用流式细胞术检测供者特异性抗体(donor specific antibodie,DSA)。在DSA最高时将供体肾脏移植至受体,构建同种异体大鼠肾移植AMR动物模型。分别于肾移植术后1~5 d收取移植肾及血清标本,行常规病理染色、C4d免疫荧光染色,以及血清DSA检测。结果:大鼠受体的血清DSA在皮肤移植术后第14天达到最高。相较于各对照组,AMR组大鼠移植肾出现显著的管周毛细血管炎和肾小球炎,免疫荧光示管周毛细血管C4d沉积显著增多,血清DSA显著升高(P < 0.05),管周毛细血管炎、肾小球炎、C4d评分显著增高(P < 0.05)。结论:成功构建并鉴定一种同种异体肾移植术后AMR的动物模型。该模型操作简便易行,稳定性和性价比较高,具有较高的推广价值。

    Abstract:

    Objective:The purpose of this resrarch is to build and identify the animal model of an antibody-mediated rejection(AMR)after kidney transplantation. Methods:In animal model of AMR,Wistar rats were used as donors and SD rats were used as the receipients. To construct allograft rat skin graft model,the back skins of Wistar rats were transplanted to the back of SD rats. After skin transplantation,serum was taken from rats at 1,3,7,10,14,21,28,and 35 days respectively,and donor specific antibodies(DSAs)of serum were detected by flow cytometry. When DSAs reached the highest,the kidney of Wistar rats was transplanted to SD rats to build AMR model of allogeneic rat kidney transplantation. Transplanted kidney and serum were collected 1-5 days after kidney transplantation,and routine pathological staining,C4d immunofluorescence and serum DSAs detection were performed. Results:DSAs of serum in recipients reached the peak after 14 days of skin transplantation. Compared with control group,transplanted kidney in AMR group showed glomerulitis with a significant increase of peritubular capillaries(PTC),C4d deposition on peritubular capillaries,and serum DSAs levels(P < 0.05). Conclusion:An animal model of AMR after renal allograft transplantation was successfully constructed and identified. This model is easy to operate,stable and cost-effective,and has high popularization value.

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程 宏,周嘉俊,王子杰,张恒诚,郑 明,韩志坚,陶 俊,居小兵,王增军,谭若芸,顾 民.肾移植术后抗体介导排斥反应模型的构建与鉴定[J].南京医科大学学报(自然科学版),2019,(5):677-681

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  • 收稿日期:2019-02-13
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  • 在线发布日期: 2019-05-31
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