对源自成人T细胞白血病（ATL）患者、同时携带ATL病毒和爱泼斯坦-巴尔病毒的人B细胞系的特性分析。

Characterization of human B-cell lines harbouring both adult T-cell leukaemia (ATL) virus and Epstein-Barr virus derived from ATL patients.

作者信息

Koyanagi Y, Yamamoto N, Kobayashi N, Hirai K, Konishi H, Takeuchi K, Tanaka Y, Hatanaka M, Hinuma Y

出版信息

J Gen Virol. 1984 Oct;65 ( Pt 10):1781-9. doi: 10.1099/0022-1317-65-10-1781.

Abstract

Human B-cell lines, designated ATLB cell lines, were spontaneously established from peripheral blood of adult T-cell leukaemia (ATL) patients. The cell lines consistently expressed ATL-associated antigen (ATLA) and Epstein-Barr virus-associated nuclear antigen (EBNA). A cloned ATLB line, designated ATLB 2, showed that both ATLA and EBNA antigens were present in the same B-cell clone. In this study, we have further characterized ATLV and EBV in the cloned ATLB 2 cell line by biochemical techniques. The ATLA antigens in these clones, initially shown by immunofluorescence, were studied by immunoprecipitation with human sera from ATL patients and the Western blotting technique using a mouse monoclonal antibody (GIN-14). We identified ATLV core polypeptides 24K and 19K in the ATLB cell extracts. Furthermore, total cellular DNA and poly(A) RNA from the ATLB cell lines were analysed for the presence of viral genomes with molecularly cloned DNA probes containing the ATLV and EBV sequence, respectively. The results showed that all ATLB 2 clones contained highly conserved ATLV proviral DNA irrespective of whether or not they expressed ATLA. They also contained several copies of EB virus DNA and DNA-DNA reassociation kinetics studies clearly showed that most of the EBV DNA sequences were present in ATLB cells. ATLV-related mRNA was detected in only ATLA-positive clones (ATLB 2-3 and 2-21) but not in a negative clone (ATLB 2-45).

摘要

从成人T细胞白血病（ATL）患者外周血中自发建立了人B细胞系，命名为ATLB细胞系。这些细胞系持续表达ATL相关抗原（ATLA）和EB病毒相关核抗原（EBNA）。一个克隆的ATLB系，命名为ATLB 2，显示ATLA和EBNA抗原存在于同一个B细胞克隆中。在本研究中，我们通过生化技术进一步对克隆的ATLB 2细胞系中的ATLV和EBV进行了特性分析。这些克隆中的ATLA抗原最初通过免疫荧光显示，随后用来自ATL患者的人血清进行免疫沉淀，并使用小鼠单克隆抗体（GIN-14）通过蛋白质印迹技术进行研究。我们在ATLB细胞提取物中鉴定出了ATLV核心多肽24K和19K。此外，分别用含有ATLV和EBV序列的分子克隆DNA探针分析了ATLB细胞系的总细胞DNA和聚腺苷酸RNA中病毒基因组的存在情况。结果表明，所有ATLB 2克隆均含有高度保守的ATLV前病毒DNA，无论它们是否表达ATLA。它们还含有多个EB病毒DNA拷贝，DNA-DNA重缔合动力学研究清楚地表明，大多数EBV DNA序列存在于ATLB细胞中。仅在ATLA阳性克隆（ATLB 2-3和2-21）中检测到了ATLV相关mRNA，而在阴性克隆（ATLB 2-45）中未检测到。