Asada H, Okada N, Hashimoto K, Yamanishi K, Sairenji T, Hirota S, Nomura S, Kitamura Y, Yoshikawa K
Department of Dermatology, Osaka University School of Medicine, Japan.
Leukemia. 1994 Aug;8(8):1415-23.
An Epstein-Barr virus (EBV) genome-positive T-cell line, designated EBT-8, was established from peripheral blood of a patient with EBV genome-positive large granular lymphocyte leukemia of T-cell origin. The cells have been cultured continuously in RPMI-1640 medium supplemented with 10% fetal calf serum and 40 U/ml interleukin-2 for more than 18 months. Analysis of T-cell receptor gene rearrangement demonstrated similar rearrangement between the fresh leukemic cells and EBT-8 cell line. The cell line has several azurophilic granules in its cytoplasm and activated cytotoxic/suppressor T-cell surface antigens (CD2, CD3, CD8, HLA-DR and T-cell receptor alpha/beta). Karyotypic analysis of the cell line showed several chromosomal abnormalities. EBV DNA was demonstrated in the cells by Southern blot hybridization and about five copies of covalently closed circular DNA per cell were detected by Gardella gel analysis. Clonotypic episomal EBV DNA was observed in the cells by Southern blot hybridization with EBV-terminal fragment probe. EBV-encoded small RNA, EBER1 were demonstrated in all cells by in situ hybridization. EBV-encoded proteins, EBNA and LMP1 were demonstrated by immunofluorescence technique. EBV activation was observed after 12-O-tetradecanoylphorbol-13- acetate treatment of the cells. These results demonstrated the establishment of a T-cell line with latent EBV genomes and suggested the involvement of EBV to the large granular lymphocyte leukemia of T cells.
从一名T细胞来源的EB病毒基因组阳性大颗粒淋巴细胞白血病患者的外周血中建立了一株EB病毒（EBV）基因组阳性T细胞系，命名为EBT - 8。这些细胞已在补充有10%胎牛血清和40 U/ml白细胞介素-2的RPMI - 1640培养基中连续培养超过18个月。T细胞受体基因重排分析表明新鲜白血病细胞与EBT - 8细胞系之间存在相似的重排。该细胞系的细胞质中有几个嗜天青颗粒，并表达活化的细胞毒性/抑制性T细胞表面抗原（CD2、CD3、CD8、HLA - DR和T细胞受体α/β）。对该细胞系的核型分析显示有几个染色体异常。通过Southern印迹杂交在细胞中检测到EBV DNA，通过Gardella凝胶分析检测到每个细胞约有五个共价闭合环状DNA拷贝。用EBV末端片段探针进行Southern印迹杂交，在细胞中观察到克隆型附加型EBV DNA。通过原位杂交在所有细胞中检测到EBV编码的小RNA，EBER1。通过免疫荧光技术检测到EBV编码的蛋白EBNA和LMP1。用12 - O - 十四酰佛波醇 - 13 - 乙酸酯处理细胞后观察到EBV活化。这些结果证明建立了一个具有潜伏性EBV基因组的T细胞系，并提示EBV与T细胞大颗粒淋巴细胞白血病有关。
