移植肾活检组织C4d免疫组化方法的比较  

COMPARATIVE STUDY FOR THE IMMUNOHISTOCHEMICAL METHODS OF C4D IN RENAL ALLOGRAFT BIOPSIES

作　　者：赵杰[1] 王慧萍[1] 陈莹[1] 冯时[1] 陈江华[1] 

机构地区：[1]浙江大学医学院附属第一医院肾脏病中心,国家中医药管理局三级实验室"肾脏病免疫实验室",杭州310003

出　　处：《中国组织化学与细胞化学杂志》2010年第3期274-279,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：115国家科技支撑计划(2008BAI60B040);浙江省科技厅重大专项(2008C13026-2)

摘　　要：目的探讨移植肾活检组织C4d免疫组化检测方法 ,寻求其实验的标准化。方法 121例移植肾活检和10例供肾标本根据ALPCO抗体手工免疫组化结果将病例分为四个组:C4d弥漫阳性组、局灶阳性组、阴性组和供肾组,采用ALPCO和ABCAM两种C4d抗体,石蜡切片行手工免疫组化和全自动免疫组化机器套染技术(以下简称机器套染),比较两种抗体和染色方法的染色结果。结果 ALPCO和ABCAM两种抗体手工免疫组化和机器套染在肾小球毛细血管基底膜(GBM)和肾小管周围毛细血管(PTC)均有良好着色,在20例ALPCO抗体手工免疫组化为弥漫阳性组标本中,染色强度在+至+++之间,而机器套染均为+,有7例标本机器套染染色强度低于手工染色,但并不改变染色的弥漫程度,另有3例其机器套染为局灶阳性;而ABCAM抗体其手工免疫组化和机器套染染色强度除1例为阴性外,其余均为+,有3例手工免疫组化和4例机器套染为局灶阳性。对于ALPCO抗体手工免疫组化为局灶阳性组的标本,其机器套染和AB-CAM抗体染色重复性较差:其机器套染有1例为阴性,而ABCAM抗体手工免疫组化有2例、机器套染有3例为阴性。ALPCO抗体手工免疫组化阴性组和供肾组标本,其机器套染和ABCAM抗体染色均为阴性,重复性良好。同时机器套染与手工免疫组化相比,拥有更好的染色背景,而且大大缩短了染色时间,更有利于急诊肾活检和免疫组化实验的标准化。结论虽然ALPCO和ABCAM两种抗体染色效果有小的差异,但总体重复性良好。ABCAM抗体在染色过程中尤其是机器套染应注意防止假阴性。手工免疫组化和机器套染联合使用为C4d免疫组化染色提供更为快捷、可靠的技术保障。Objective To investigate the standard immunohistochemical detection methods of C4d in renal allograft biopsies. Methods We used ALPCO and ABCAM antibodies to detect 121 cases of renal allograft biopsies and 10 cases of donor kidney by manual and manchine automated immunohistochemical staining, and then compared the results of staining.According to the results of ALPCO manual staining,the cases were divided into 4 groups： C4d diffuse-positive, C4d focal-positive, negative, and donor kidney.Results ALPCO and ABCAM polyclonal antibodies had good staining results in the glomerular capillary basement membrane （GBM） and peritubular capillary （PTC）.In 20 cases of the diffuse-positive group by ALPCO manual immunohistochemical method,the staining intensity changed from ＋ to ＋＋＋,but only ＋ in manchine automated immunohistochemical staining.There were 7 cases whose intensity of machine staining was lower than manual,but this did not change the degree of the diffuse staining.Besides,machine staining showed 3 cases to be focal-positive.The staining intensity of ABCAM were all ＋ except one negative case,and ABCAM showed 3 in manual staining and 4 in machine staining to be focal-positive.In the ALPCO manual focal-positive group, machine staining showed 1 negative case and ABCAM showed 2 negative cases in manual,and 3 in machine staining,so machine staining and ABCAM had poor repeatability.In the negative and donor kidney groups by ALPCO manual staining,its machine and ABCAM methods had negative results.Comparing machine and manual staining,the machine had better staining background and greatly shortened the staining time,especially in emergency renal allograft biopsies.Conclusion The two antibodies have good repeatability although they have some differences,and in the machine staining of ABCAM we should avoid false-negtive results.We can combine manual with automated machine staining methods of C4d and gain much more efficient and reliable technical support.

关 键 词：C4D 移植肾活检 肾小管周围毛细血管 手工免疫组化 机器套染 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]