体内衍生的硫鸟嘌呤抗性人B细胞的克隆

Cloning of in vivo-derived thioguanine-resistant human B cells.

作者信息

Hakoda M, Hirai Y, Kusunoki Y, Akiyama M

机构信息

Department of Radiobiology, Radiation Effects Research Foundation, Hiroshima, Japan.

出版信息

Mutat Res. 1989 Jan;210(1):29-34. doi: 10.1016/0027-5107(89)90041-9.

DOI:10.1016/0027-5107(89)90041-9

PMID:2535889

Abstract

In vivo-derived thioguanine-resistant (TGr) B cells have been cloned from peripheral blood mononuclear cells (PBMC) of 4 healthy adults. This was done by using Epstein-Barr (EB) virus transformation of B cells enriched from a large number of PBMC obtained with a blood cell separator. The cloned TGr B cells lacked hypoxanthine guanine phosphoribosyltransferase (HPRT) enzyme activity. The frequency of in vivo TGr B cells was estimated to be 8.6-13.1 X 10(-6) for the 4 individuals by comparing the cloning efficiency of non-selected cells and TG-selected cells. This frequency is somewhat higher but comparable to the in vivo frequency of TGr T cells. Because the cloned TGr B cells can be easily expanded in vitro, this procedure provides a large amount of material for the precise characterization of in vivo mutations in humans.

摘要

已从4名健康成年人的外周血单个核细胞（PBMC）中克隆出体内衍生的硫鸟嘌呤抗性（TGr）B细胞。这是通过用爱泼斯坦-巴尔（EB）病毒转化从血细胞分离器获得的大量PBMC中富集的B细胞来完成的。克隆的TGr B细胞缺乏次黄嘌呤鸟嘌呤磷酸核糖转移酶（HPRT）酶活性。通过比较未选择细胞和TG选择细胞的克隆效率，估计这4名个体体内TGr B细胞的频率为8.6 - 13.1×10^(-6)。该频率略高，但与体内TGr T细胞的频率相当。由于克隆的TGr B细胞可以在体外轻松扩增，该方法为精确表征人类体内突变提供了大量材料。