我国3例范可尼贫血患者原始B-淋巴细胞系的建立和分型  

Establishment of EBV-immortalized lymphoblast cell lines from three Chinese Fanconi anemia patient and their subtyping

作　　者：谢燕[1] Hans Joenje 彭光洁[3] 陈友华[1] 陈伦华[1] 

机构地区：[1]湖北医科大学附属第一医院血液科,武汉市430060 [2]Department of Human Genetics [3]同济医科大学附属同济医院儿科

出　　处：《中华血液学杂志》2001年第4期173-176,共4页Chinese Journal of Hematology

摘　　要：目的　通过建立我国范可尼贫血 (Fanconianemia,FA)患者原始B 淋巴细胞系进行FA分型。方法　将我国 3例FA患者外周血细胞用EB病毒活化法建立原始B 淋巴细胞系 ,采用丝裂霉素(MMC)生长抑制试验和细胞融合后MMC抑制试验的互补试验进行分型 ;利用单克隆抗体免疫沉淀法和Westernblot法进行分型。结果　成功建立了 3个FA原始B 淋巴细胞系 ,其中 2个细胞系仍然保持对MMC的敏感性 ,IC50 均小于 10nmol/L ;用 1例嵌合体患者细胞所建立的原始B 淋巴细胞系对MMC不敏感 ,IC50 大于 10nmol/L ;经MMC抑制试验的互补试验和Westernblot检测 ,3例FA患者均为FA A亚型。结论　FA两种分型方法各有利弊 ,细胞融合后MMC生长抑制试验分型可靠 ,适用于所有未知FA亚型的患者 ,但该法依赖于建立MMC敏感的原始B 淋巴细胞系 ,无法用于FA A嵌合体患者 ,方法耗时、耗力 ,出结果慢 ;而利用单克隆抗体免疫沉淀法和Westernblot法分型不依赖于建立MMC敏感的原始B 淋巴细胞系 ,可用于FA嵌合体患者 ,但仅适用于基因已知和已有特异性抗体的亚型 ,方法简单 ,出结果快。Objective To Establish EBV immortalized lymphoblast cell lines from Chinese Fanconi anemia(FA) patients. Methods The establishment of lymphoblast cell lines was by EBV immortalization and subtyping of the cell lines by cell fusion,mytomycin C (MMC) inhibition analysis and /or immunoprecipitation and Western blot. Result ①Three lymphoblast cell lines were established,two of them were sensitive to MMC with a IC 50 <10?nmol/L.②The subtypes of all the three patients belong to FA A group. Conclusion Complementation analysis was suitable for all FA patients with MMC sensitive lymphoblastic cell lines, but this method is time consuming. Immunoprecipitation and Western blot are much faster for subtyping but could only be used for FA patients whose related genes had been cloned and specific antibodies been developed.

关 键 词：范可尼贫血 细胞融合 沉淀素试验 B淋巴细胞系 建立 分型 

分 类 号：R556.9[医药卫生—血液循环系统疾病]