供体来源细胞游离DNA鉴别移植肾急性排斥反应类型  

Donor-derived cell-free DNA can discriminate acute rejection types after kidney transplantation

作　　者：程雅妹 郭陆英 雷文华 吕军好 严芃芃 沈佳 王美芳 周芹 王慧萍 陈江华 王仁定 Cheng Yamei;Guo Luying;Lei Wenhua;Lyu Junhao;Yan Pengpeng;Shen Jia;Wang Meifang;Zhou Qin;Wang Huiping;Chen Jianghua;Wang Rending(Kidney Disease Center,the First Affiliated Hospital,College of Medicine,Zhejiang University,Key Laboratory of Kidney Disease Prevention and Control Technology,Zhejiang Province,Zhejiang University Institute of Nephrology,Hangzhou 310003,China;Organ Donation and Coordination Office,the First Affiliated Hospital,College of Medicine,Zhejiang University,Hangzhou 310003,China)

机构地区：[1]浙江大学医学院附属第一医院肾脏病中心,浙江省肾脏病防治技术研究重点实验室,浙江大学肾脏病研究所,杭州310003 [2]浙江大学医学院附属第一医院人体器官捐献协调办公室,杭州310003

出　　处：《中华肾脏病杂志》2022年第1期32-38,共7页Chinese Journal of Nephrology

基　　金：国家自然科学基金(81870510);浙江省科技厅重点研发项目基金(2019C03029);白求恩慈善基金(G-X-2019-0101-12)。

摘　　要：目的探讨血浆供体来源的细胞游离DNA(donor-derived cell-free DNA,dd-cfDNA)片段在鉴别移植肾T细胞介导的排斥反应(T cell-mediated rejection,TCMR)和抗体介导的排斥反应(antibody-mediated rejection,ABMR)中的价值。方法回顾性纳入2017年12月1日至2019年7月18日在浙江大学医学院附属第一医院经肾活检确诊的移植肾急性排斥反应患者。根据2017年Banff移植肾排斥反应病理分类诊断标准将患者分为ABMR组和TCMR组,后者又分为TCMRⅠ型亚组和TCMRⅡ型亚组。采用二代测序+目标区域捕获的方法检测入选者外周血浆dd-cfDNA水平。比较两组患者人口学及临床病理指标的差异。采用受试者工作特征曲线(receiver operating characteristic curve,ROC曲线)评价血浆dd-cfDNA和血肌酐水平对鉴别两种移植肾ABMR和TCMR的诊断价值。结果共60例移植肾急性排斥反应患者入选本研究,TCMR组42例,ABMR组18例。ABMR组患者血浆dd-cfDNA百分比较TCMR组显著升高[2.33(1.19,4.30)%比0.98(0.50,1.82)%,P=0.001];ABMR组dd-cfDNA绝对值亦较TCMR组显著升高[0.94(0.60,2.27)ng/ml比0.43(0.20,0.96)ng/ml,P=0.003]。ROC曲线分析结果显示,dd-cfDNA百分比鉴别诊断TCMR和ABMR的曲线下面积(area under the curve,AUC)为0.76(95%CI 0.64~0.88),阈值为1.11%,敏感性88.89%,特异性59.52%;dd-cfDNA绝对值鉴别诊断TCMR和ABMR的AUC为0.74(95%CI 0.61~0.86),阈值为0.53 ng/ml,敏感性88.89%,特异性54.76%。TCMR亚组与ABMR组血浆dd-cfDNA水平的比较结果显示,TCMR两亚组间dd-cfDNA百分比及绝对值水平的差异均无统计学意义(均P>0.05);ABMR组dd-cfDNA百分比显著高于TCMRⅠ型亚组(P=0.008)和TCMRⅡ型亚组(P=0.030)。ABMR组dd-cfDNA绝对值显著高于TCMRⅠ型亚组(P=0.003)。结论血浆dd-cfDNA水平检测对鉴别ABMR和TCMR两种排斥反应类型有辅助诊断价值。Objective To explore the value of detecting plasma donor-derived free DNA(dd-cfDNA)fraction in distinguishing antibody mediated-rejection(ABMR)and T cell-mediated rejection(TCMR)of renal allografts.Methods Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1,2017 to July 18,2019 were retrospectively included.Based on pathological classification of Banff renal allograft rejection in 2017,the patients were divided into ABMR group and TCMR group,and the latter was subdivided into TCMRⅠ subgroup and TCMRⅡ subgroup.The second generation sequencing and target region capture were used to detect candidates'peripheral blood dd-cfDNA.The demographic and clinicopathological data of the two groups were compared.The receiver operating characteristic curve(ROC)was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results A total of 60 patients with acute rejection of renal transplantation were enrolled in this study,including 42 patients in TCMR group and 18 patients in ABMR group.The plasma dd-cfDNA percentage(%)in the ABMR group was significantly higher than that in the TCMR group[2.33(1.19,4.30)%vs 0.98(0.50,1.82)%,P=0.001].The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group[0.94(0.60,2.27)ng/ml vs 0.43(0.20,0.96)ng/ml,P=0.003].ROC analysis to discriminate TCMR from ABMR showed that,the area under the curve(AUC)of dd-cfDNA% was 0.76(95%CI 0.64-0.88),when the threshold was 1.11%,the sensitivity and specificity were 88.89% and 59.52%,respectively;the AUC of absolute value of dd-cfDNA was 0.74(95%CI 0.61-0.86),when the threshold was 0.53 ng/ml,the sensitivity was 88.89% and the specificity was 54.76%.TCMR subgroups were further analyzed,there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA(both P>0.05);dd-cfDNA% in ABMR group was apparently higher than

关 键 词：肾移植 移植物排斥 鉴别 诊断 供体来源细胞游离DNA 

分 类 号：R699.2[医药卫生—泌尿科学]