同种异体小鼠肾移植慢性抗体介导排斥反应模型的构建与鉴定
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1南京医科大学第二附属医院泌尿外科,江苏 南京 210011 ; 2.南京医科大学第一附属医院泌尿外科,江苏 南京 210029

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国家自然科学基金(82170769,82470790)


Construction and identification of a chronic antibody ⁃ mediated rejection model of renal graft in mice
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1Department of Urology,the Second Affiliated Hospital of Nanjing Medical University,Nanjing 210011 ; 2.Departmentof Urology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029 ,China

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    摘要:

    目的:构建并鉴定同种异体小鼠肾移植慢性抗体介导排斥反应(chronic antibody-mediated rejection,cABMR)模型。 方法:采用雄性6~8周龄BALB/cH2d 与C57BL/6H2b 小鼠,随机分为:①同基因对照组;②细胞介导的排斥反应组;③急性抗体介导排斥反应组(acute antibody-mediated rejection,aABMR);④cABMR 组1;⑤cABMR 组2;⑥cABMR 组3。移植肾行病理染色及 Banff评分诊断,绘制生存曲线,检测移植肾组织中补体片段C3d沉积、外周血IgG类供体特异性抗体(donor specific antibody, DSA)、血清肌酐(creatinine,Cr)、血清尿素氮(blood urea nitrogen,BUN)水平,检测移植肾组织中CD3+ T细胞、CD19+ B细胞、F4/80标记的巨噬细胞、Ly6G 标记的髓系细胞数量。结果:cABMR 组 1 模型符合抗体介导排斥反应(antibody-mediated rejection, ABMR)诊断标准,术后存活率为80%,CD3+ T细胞不明显,故选用cABMR组1作为cABMR模型进行后续论证。与对照组相比, cABMR的C3d表达阳性区域显著增多,外周血IgG类DSA平均荧光强度明显增强,外周血血清Cr与BUN水平明显升高。与 aABMR相比,cABMR移植肾管周毛细血管出血、扩张,肾小球内可见单个核细胞浸润,肾小管内出现更典型的小管损伤及刷状缘脱落,Banff评分更高,C3d表达阳性区域显著增多,外周血IgG类DSA平均荧光强度显著增强,外周血血清Cr与BUN水平明显升高。与aABMR相比,cABMR肾间质和管周毛细血管CD19+ B细胞、CD3+ T细胞浸润均不显著,肾间质F4/80标记的巨噬细胞、Ly6G标记的髓系细胞浸润均更显著,Foxp3与白介素(interleukin,IL)-10的表达在cABMR模型中显著减少,IL-1β、IL-6、 肿瘤坏死因子α(tumor necrosis factor α,TNF-α)、趋化因子CCL2炎症细胞因子的浸润在cABMR模型中显著增多。结论:成功构建并鉴定一种生存周期得到延长的cABMR模型,可在此模型的基础上评估cABMR的发生机制及干预措施,具有较高的临床应用价值。

    Abstract:

    Objective:Construction and identification of a chronic antibody - mediated rejection(cABMR)model of renal graft in mice. Methods:Male 6-8 weeks old BALB/cH2d and C57BL/6H2b mice were randomly divided into the syngeneic control group,cell - mediated rejection group,acute antibody - mediated rejection(aABMR)group,cABMR group 1,cABMR group 2,cABMR group 3. Pathological staining and Banff score were performed to diagnose the transplanted kidneys,survival curves were plotted,the levels of complement fragment C3d deposition,peripheral blood IgG class donor-specific antibody(DSA),serum creatinine(Cr)and serum urea nitrogen(BUN)were detected in transplanted kidney tissues,the expression of CD3 + T cells,CD19 + B cells,F4/80 labelled macrophages,Ly6G labelled myeloid cells were detected. Results:cABMR group 1 met ABMR diagnostic criteria with 80% postoperative survival rate and negligible CD3+ T cell expression,thus selected for subsequent studies. Compared to the control group,the C3d - positive exoression regions were significantly increased in the cABMR group and the average fluorescence intensity of IgG DSA in peripheral blood was notably enhanced. Serum Cr and BUN levels in peripheral blood were significantly higher. Compared to aABMR,the cABMR model of transplanted kidneys showed more extensive peritubular capillary haemorrhage and dilatation,single nucleus infiltration in the glomeruli,and more typical tubular damage and brush border detachment in the tubules,higher Banff scores, significantly more positive areas of C3d expression,significantly higher mean fluorescence intensity of peripheral blood IgG DSA, significantly higher peripheral blood serum Cr and BUN levels. Incontrast to aABMR,cABMR group showed no siginificant infiltration of CD19+ B cells or CD3+ T cells in the renal interstitial or peritubular capillaries,but there was a more significant infiltration of renal interstitial F4/80 labelled macrophage and Ly6G labelled myeloid cell. Foxp3 and IL-10 expression were significantly reduced in the cABMR model,while the infiltration of IL-1β,IL-6,TNF-α and CCL2 inflammatory cytokines was significantly increased in cABMR model. Conclusion:A cABMR model with an extended survival period was successfully constructed and identified. This model can be used to assess the mechanisms and intervention strategies of cABMR,with high clinical application value.

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喻琦钦,杨泽昕,张俊麒,桂泽平,倪斌,郑明,沈百欣,王子杰,顾民.同种异体小鼠肾移植慢性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),2025,(8):1073-1081

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  • 收稿日期:2025-01-23
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  • 在线发布日期: 2025-08-13
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