Trivedi P, Cuomo L, de Campos-Lima P O, Imreh M P, Kvarnung K, Klein G, Masucci M G
Department of Tumour Biology, Karolinska Institute, Stockholm, Sweden.
J Gen Virol. 1993 Jul;74 ( Pt 7):1393-8. doi: 10.1099/0022-1317-74-7-1393.
We have analysed the expression of transformation-associated viral antigens, the Epstein-Barr virus (EBV) DNA content and the phenotypic characteristics of two B95-8 virus-converted sublines of the EBV-negative Burkitt's lymphoma (BL) line BL28. The converted lines called E95A-BL28 and E95B-BL28, respectively, differed in their EBV gene expression. The E95B convertant expressed virus-encoded nuclear antigens EBNA1 to -6 and the membrane protein LMP1, but only EBNA2 and EBNA5 were detected by immunofluorescence and immunoblotting in the E95A convertant. Only the entire BamHI W, Y and H regions could be detected in the E95A convertant by hybridization of Southern blots with probes covering the BamHI C, W, Y, H, F, E, K and Nhet regions of the EBV genome. EBV episomes were found to be absent in the E95A convertant as seen by Gardella gels. The E95A convertant retained the phenotypic characteristics of the EBV-negative parental line, and remained highly clonable in agarose. In contrast, expression of EBNA1 to -6 and LMP1 was accompanied by a shift towards a more lymphoblastoid cell line-like phenotype and by loss of agarose clonability in the E95B convertant.
我们分析了EB病毒阴性的伯基特淋巴瘤（BL）细胞系BL28的两个B95 - 8病毒转化亚系中转化相关病毒抗原的表达、EB病毒（EBV）DNA含量及表型特征。这两个分别称为E95A - BL28和E95B - BL28的转化细胞系，其EBV基因表达存在差异。E95B转化体表达病毒编码的核抗原EBNA1至 - 6及膜蛋白LMP1，但在E95A转化体中通过免疫荧光和免疫印迹仅检测到EBNA2和EBNA5。通过用覆盖EBV基因组BamHI C、W、Y、H、F、E、K和Nhet区域的探针进行Southern印迹杂交，在E95A转化体中仅能检测到完整的BamHI W、Y和H区域。从Gardella凝胶分析可见，E95A转化体中不存在EBV附加体。E95A转化体保留了EBV阴性亲代细胞系的表型特征，并且在琼脂糖中仍具有高度可克隆性。相比之下，E95B转化体中EBNA1至 - 6和LMP1的表达伴随着向更类似淋巴母细胞系表型的转变以及琼脂糖可克隆性的丧失。
