Hoshino H, Esumi H, Miwa M, Shimoyama M, Minato K, Tobinai K, Hirose M, Watanabe S, Inada N, Kinoshita K, Kamihira S, Ichimaru M, Sugimura T
Proc Natl Acad Sci U S A. 1983 Oct;80(19):6061-5. doi: 10.1073/pnas.80.19.6061.
By using human T-cell growth factor (TCGF), 10 cell lines were established from tissue samples of 10 patients with adult T-cell leukemia (ATL). Three cell lines were adapted to growth in medium lacking TCGF. The surface markers of all cell lines were characteristic of inducer/helper T cells, i.e., OKT3+, OKT4+, OKT6-, OKT8-, OKIa1+, and human Lyt2+ and Lyt3+, except that one cell line was OKT3-. The expression of the viral antigen was examined during establishment of 8 of the 10 cell lines. The viral antigen was not expressed in leukemic cells before cultivation. In 5 lines, the viral antigen was detected by immunofluorescent staining after a short period of cultivation. However, 3 cell lines, ATL-6A, ATL-9Y, and ATL-1K did not express the viral antigen during short-term culture: the ATL-6A and ATL-9Y cell lines became positive for the viral antigen after 5 and 2 months of cultivation, respectively; the ATL-1K cell line remained antigen-negative throughout a culture period of 13 months. Southern blot hybridization assay showed that all of the cell lines, including the viral antigen-negative ATL-1K cell line, contained the viral genome. Thus, the retrovirus was associated with all 10 cell lines established from ATL patients, but there was a heterogeneity in the expression time of the retroviral antigen in leukemic cells maintained in vitro. Our findings suggested that the expression of the viral antigen was not required for maintenance of the leukemic state in vivo and for growth of leukemic cells in vitro.
通过使用人T细胞生长因子(TCGF),从10例成人T细胞白血病(ATL)患者的组织样本中建立了10个细胞系。其中3个细胞系适应了在不含TCGF的培养基中生长。所有细胞系的表面标志物均为诱导/辅助性T细胞的特征,即OKT3 +、OKT4 +、OKT6 -、OKT8 -、OKIa1 +以及人Lyt2 +和Lyt3 +,但有一个细胞系为OKT3 -。在10个细胞系中的8个细胞系建立过程中检测了病毒抗原的表达。培养前白血病细胞中未表达病毒抗原。在5个细胞系中,短时间培养后通过免疫荧光染色检测到病毒抗原。然而,3个细胞系ATL - 6A、ATL - 9Y和ATL - 1K在短期培养期间未表达病毒抗原:ATL - 6A和ATL - 9Y细胞系分别在培养5个月和2个月后病毒抗原呈阳性;ATL - 1K细胞系在13个月的培养期内一直呈抗原阴性。Southern印迹杂交分析表明,所有细胞系,包括病毒抗原阴性的ATL - 1K细胞系,均含有病毒基因组。因此,逆转录病毒与从ATL患者建立的所有10个细胞系相关,但体外培养的白血病细胞中逆转录病毒抗原的表达时间存在异质性。我们的研究结果表明,病毒抗原的表达对于体内白血病状态的维持和体外白血病细胞的生长并非必需。