Drover S, Marshall W H
Immunology Laboratory, Faculty of Medicine, Memorial University of Newfoundland, St. John's Canada.
Hum Immunol. 1991 Aug;31(4):293-8. doi: 10.1016/0198-8859(91)90102-f.
Mouse L cells expressing HLA genes are potentially useful for producing and analyzing monoclonal antibodies (mAb) to HLA molecules. This paper describes the preparation of transfectants using uncloned human DNA and three methods to isolate the HLA-expressing transfectants. Transfectant libraries were made by cotransfecting mouse thymidine kinase (tk)-deficient L cells with a calcium phosphate precipitate containing genomic DNA and tk plasmid DNA. Transfectants expressing HLA genes were isolated using these methods: immunomagnetism, replicate-plating combined with cellular enzyme-linked immunoassay, and sorting using a fluorescence-activated cell sorter. Two HLA-A2 transfectant were isolated using immunomagnetism, two HLA-A24 transfectants by replicate plating, and one HLA-Bw60 transfectant by FACS. However, no transfectants were isolated that stably expressed class II genes. The class I transfectants have been useful in characterizing several locally prepared mAbs which bind to monomorphic determinants on class I HLA molecules. Two of the transfectant lines, one expressing HLA-A2 (8001) and the other HLA-A24 (8008), have been included in the collection of lines distributed for use in the Eleventh International Histocompatibility Workshop.
表达HLA基因的小鼠L细胞在生产和分析针对HLA分子的单克隆抗体（mAb）方面具有潜在用途。本文描述了使用未克隆的人类DNA制备转染子以及三种分离表达HLA的转染子的方法。通过用含有基因组DNA和tk质粒DNA的磷酸钙沉淀物共转染小鼠胸苷激酶（tk）缺陷型L细胞来构建转染子文库。使用以下方法分离表达HLA基因的转染子：免疫磁珠法、结合细胞酶联免疫测定的复制铺板法以及使用荧光激活细胞分选仪进行分选。使用免疫磁珠法分离出两个HLA-A2转染子，通过复制铺板法分离出两个HLA-A24转染子，通过荧光激活细胞分选仪（FACS）分离出一个HLA-Bw60转染子。然而，未分离出稳定表达II类基因的转染子。I类转染子在鉴定几种本地制备的与I类HLA分子上的单态性决定簇结合的单克隆抗体方面很有用。其中两个转染子系，一个表达HLA-A2（8001），另一个表达HLA-A24（8008），已被纳入为第十一届国际组织相容性研讨会分发使用的细胞系集合中。
