ELISPOT技术检测供体特异性IFN-γ分泌频数在早期诊断急性排斥反应中的意义  

The significance of detecting donor-specific IFN-γ-secreting lymphocytes by ELISPOT assay in early diagnosis of acute rejection

作　　者：朱鹏[1] 陈义发[1] 陈孝平[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科中心,武汉430030

出　　处：《中华微生物学和免疫学杂志》2007年第9期860-863,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金项目(30571754);留学回国人员科研启动基金[教外司留(2005)383]

摘　　要：目的探讨急性排斥反应的早期诊断方法及ELISPOT技术在供体评估方面的应用价值。方法采用小鼠腹部心脏移植模型，分为移植排斥组、实验处理组和同系移植组，每组20对。观察移植物存活时间和供心病理改变，应用ELISPOT技术检测受鼠脾细胞悬液IFN-γ活性分泌细胞频数。结果移植排斥组及实验处理组移植物平均存活时间分别为（7．80±0．77）d、（14．80±1．01）d，同系移植组移植物存活均超过28d，3组差异有统计学意义。移植排斥组与实验组及同系移植组相比较，移植物内心肌细胞变性坏死严重并有大量炎性细胞浸润。移植术后第7天，3组供体特异性IFN-γ活性分泌细胞频数分别为（416±19）、（44±8）、（7±2）个／2×10^5，其与相应移植物存活时间存在明显负相关，而与病理分级存在明显正相关。而非特异性IFN-γ活性分泌细胞频数均很低，差异无统计学意义。结论应用ELISPOT技术检测术后早期供体特异性IFN-γ活性分泌细胞频数可作为急性排斥反应的一个早期诊断指标。此技术具有较高的灵敏性与特异性，可用于术前供体配型。Objective To develop a method for early diagnosis of acute rejection and the significance of ELISPOT in donor evaluation. Methods All ventral heterotopic cardiac transplantation models were divided into three groups as rejection group, treated group and isograft group. Mean survival time （MST） and pathological changes were measured. With ELISPOT assay, IFN-γ-secreting lymphocytes were detected and counted. Results MST of heart allografts in rejection and treated group were （7.80 ± 0.77） d, （ 14.80 ± 1.01 ） d, respectively. The survival time of grafts in isograft group was all more than 28 d. The difference among three groups was obvious. The number of infiltrating cells in rejection group was much more than the other groups as well as the extent of pathological changes. The numbers of donor-specific IFN-γ-secreting lymphocytes in three groups were （416 ± 19）, （44 ± 8）, （7 ± 2）/2 × 10^5 , respectively, which had negative correlation with MST and positive correlation with pathological changes. Conclusion The detection of donor-specific IFN-γ-secreting lymphocytes with ELISPOT assay may be a useful indicator for early diagnosis of acute rejection. This assay has enough sensitivity and specificity for pre-transplant donor evaluation.

关 键 词：酶联免疫斑点技术 Γ干扰素 心脏移植 移植物 小鼠 

分 类 号：R446.6[医药卫生—诊断学]