流式淋巴毒技术在预致敏肾移植中的应用初探  

Pilot use of Flow-CDC method during pre-sensitized kidney transplantation

作　　者：丁韬 李宁 王璐[3] 冯豪 郭志良 朱兰[3] 陈刚[3] Ding Tao;Li Ning;Wang Lu;Feng Hao;Guo Zhiliang;Zhu Lan;Chen Gang(Department of Organ Transplantation,Dongfeng General Hospital,Hubei University of Medicine,Shiyan 442000,China;Department of Pediatric Surgery,Tongji Hospital,Huazhong University of Science and Technology,Wuhan 430030,China;Institute of Organ Transplantation,Tongji Hospital,Huazhong University of Science and Technology/Key Laboratory of Organ Transplantation,Ministry of Education,China/Key Laboratory of Organ Transplantation,Ministry of Public Health,Wuhan 430030,China)

机构地区：[1]湖北医药学院附属东风医院器官移植科,十堰442000 [2]华中科技大学同济医学院附属同济医院儿科,武汉430030 [3]华中科技大学同济医学院附属同济医院器官移植研究所教育部/卫生健康委员会/中国医学科学院器官移植重点实验室,武汉430030

出　　处：《中华器官移植杂志》2019年第12期712-716,共5页Chinese Journal of Organ Transplantation

基　　金：2017年湖北省科技计划项目(2017ACA096)。

摘　　要：目的建立一种对于预致敏肾移植行之有效而特异敏感的流式淋巴毒(Flow-CDC)检测技术,并通过实例说明其在临床中的应用价值。方法经Luminex提示预存供者特异性抗体(DSA)阳性的10例再次和多次肾移植受者进行术前免疫评估。供者淋巴细胞与受者血清孵育后加入兔补体,以碘化丙啶(PI)标记死细胞,用流式细胞仪检测死亡细胞百分比。7例为传统CDC/Flow-CDC均(-),接受器官捐献供肾移植;2例为传统CDC(-)/Flow-CDC(+),经1~2周去敏治疗至Flow-CDC转阴后接受亲属活体供肾移植;1例为传统CDC(+)/Flow-CDC(-),经1次急诊血浆置换后接受器官捐献供肾移植。结果对于群体反应性抗体(PRA)为不同程度阳性的血清,Flow-CDC能准确检测出不同定量水平的淋巴细胞杀伤率。10例预存Luminex-DSA阳性受者均未出现超急性排斥反应,术后1个月内发生急性混合性排斥反应2例,均治疗后痊愈。结论Flow-CDC检测准确可靠,值得在预致敏肾移植的术前免疫评估中积极应用。Objective To establish an effective and sensitive method for detecting antibodies with complement-dependent lymphocyte toxicity by flow cytometry(Flow-CDC)in pre-sensitized renal graft recipients and illustrate its application with samples from clinical cases.Methods Donor lymphocytes were incubated with recipient sera in the presence of rabbit complement.Dead cells were labeled with propidium iodide(PI)while the percentage of dead cells was measured by flow cytometry.This method was employed for preoperative immunological risk evaluations in 10 renal re-transplant recipients with high-level of pre-existing donor-specific antibodies(DSA)based upon Luminex.Seven patients with both negative CDC and Flow-CDC tests received kidney transplantation directly from potential deceased donors.Two patients with negative CDC but positive Flow-CDC had renal transplantation from their living-donors in 1 to 2 weeks later when Flow-CDC became negative after a desensitization therapy.With a negative Flow-CDC,one patient had a positive CDC to a potential deceased donor.Kidney transplantation was performed after one session of preoperative plasmapheresis.Results For sera with different levels of panel reactive antibodies,mortality rate of target lymphocytes may be accurately evaluated by Flow-CDC test.No hyperacute rejection occurred in 10 cases of positive pre-existing Luminex DSA.Two cases of mixed acute rejection within 1 month post-transplantation recovered after treatment.Conclusions Flow-CDC is an accurate and reliable test suitable for preoperative immunological evaluations of patients with sensitization history.

关 键 词：肾移植 HLA配型 流式细胞术 

分 类 号：R699[医药卫生—泌尿科学]