Casareale D, Stevenson M, Sakai K, Volsky D J
Virology. 1987 Jan;156(1):40-9. doi: 10.1016/0042-6822(87)90434-x.
Infection of human helper T lymphocytes with the human immunodeficiency virus (HIV) results in a rapid induction of cytopathic effects and cell lysis. We isolated a variant of the human T-lymphoblastoid cell line, CEM, that is fully susceptible to HIV infection but resistant to virally induced cytopathic effects. Exposure of the cells, designated CR-10, to HIV resulted in the expression of viral antigens in 100% of cells within 6-9 days. Virus-infected cells remained fully viable and could be cultivated under standard culture conditions for a desired period of time. Parental CEM cells died within 9-12 days after HIV infection. Proviral DNA could be detected in the HIV-infected CR-10 cells by Southern blot and molecular hybridization 4-5 days after infection; the relative amount of proviral DNA reached maximum at Days 6-10 and remained stable during an 8-month follow-up period. Virus production by HIV-infected CR-10 cells was documented by electron microscopy and detection of reverse transcriptase activity in cell culture supernatants. HIV-infected CR-10 cells exhibited a down modulation of the OKT-3, OKT-4, OKT-4A, OKT-8, and OKT-11 T-cell surface markers, but not of the OKT-9 (transferrin receptor). One of the HIV persistently infected CR-10 cell clones has been kept in continuous culture for over 8 months. During this period, the cells remained fully viable, 100% positive for HIV antigens, and negative for most of the T-cell surface markers tested and continued to produce biologically active HIV. The CR-10 and HIV-infected CR-10 cell lines will be useful in studies on the biology of HIV and in the isolation and large-scale propagation of this virus.
人类免疫缺陷病毒(HIV)感染人类辅助性T淋巴细胞会迅速引发细胞病变效应和细胞裂解。我们分离出了人T淋巴母细胞系CEM的一个变体,它对HIV感染完全敏感,但对病毒诱导的细胞病变效应具有抗性。将命名为CR-10的细胞暴露于HIV后,在6 - 9天内100%的细胞中都会表达病毒抗原。被病毒感染的细胞仍完全存活,并可在标准培养条件下培养所需时间。亲代CEM细胞在HIV感染后9 - 12天内死亡。感染后4 - 5天,通过Southern印迹和分子杂交可在HIV感染的CR-10细胞中检测到前病毒DNA;前病毒DNA的相对量在第6 - 10天达到最大值,并在8个月的随访期内保持稳定。通过电子显微镜和检测细胞培养上清液中的逆转录酶活性,证明了HIV感染的CR-10细胞产生病毒。HIV感染的CR-10细胞表现出OKT-3、OKT-4、OKT-4A、OKT-8和OKT-11 T细胞表面标志物的下调,但OKT-9(转铁蛋白受体)没有下调。其中一个持续感染HIV的CR-10细胞克隆已连续培养超过8个月。在此期间,细胞仍完全存活,HIV抗原呈100%阳性,所检测的大多数T细胞表面标志物呈阴性,并继续产生具有生物活性的HIV。CR-10细胞系和HIV感染的CR-10细胞系将有助于HIV生物学研究以及该病毒的分离和大规模繁殖。