HLA antigen expression at the single cell level on a K562 X B cell hybrid: an analysis with monoclonal antibodies using bacterial binding assays.

A Ziegler, B Uchańska-Ziegler, J Zeuthen, P Wernet
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引用次数: 81

Abstract

The influence of different genetic environments on the expression of HLA complex-controlled antigens has been investigated using cell lines with various defects in the synthesis of these molecules and a somatic cell hybrid derived from them. A very sensitive bacterial binding assay allowing simultaneous evaluation of the morphology of a given cell and the quantity of a surface molecule has been developed for these studies. The fetal erythroid cell line K562, the Burkitt's lymphoma-derived cell line DAUDI, and their hybrid DUTKO1 have been employed. K562 and the hybrid, but not DAUDI, expressed HLA-A,B,C heavy chains as detected by the monoclonal antibody W6/32.HL, while two monoclonal antibodies (TU48 and 2BC4) against the supertypic specificities HLA-Bw4 and Bw6 showed no reactivity. The presence of human Ia-like antigens on the cell surfaces was investigated with a panel of eight monoclonal antibodies. K562 cells were completely unreactive, and DAUDI cells gave the expected positive reaction, but about 1% or less of the cells in the DUTKO1 population appeared to express these antigens as well. We discuss possible reasons for the failure to detect HLA-B antigens with monoclonal antibodies and the lack of complete "dominance" of the K562 genome in the hybrid cell line.

HLA抗原在K562 X B细胞杂交株单细胞水平上的表达:单克隆抗体细菌结合分析
不同的遗传环境对HLA复合物控制抗原表达的影响已经被研究,使用在这些分子的合成中有各种缺陷的细胞系和由它们衍生的体细胞杂交。一种非常敏感的细菌结合试验允许同时评估给定细胞的形态和表面分子的数量,已经为这些研究开发。采用胎红细胞系K562、伯基特淋巴瘤衍生细胞系DAUDI及其杂交细胞系DUTKO1。单克隆抗体W6/32检测到K562和杂交株表达HLA-A、B、C重链,而DAUDI不表达。而针对超型特异性HLA-Bw4和Bw6的两种单克隆抗体(TU48和2BC4)无反应性。用8种单克隆抗体检测人ia样抗原在细胞表面的存在。K562细胞完全无反应,DAUDI细胞表现出预期的阳性反应,但DUTKO1群体中约1%或更少的细胞也表达这些抗原。我们讨论了用单克隆抗体检测HLA-B抗原失败的可能原因,以及在杂交细胞系中缺乏K562基因组的完全“优势”。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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