INDUCTION OF HIV-1 FROM LATENTLY INFECTED CELL LINES BY CHEMICAL CARCINOGENS

来自慢性感染的U937培养物的U1细胞株含有 在TPA、TNFa、IL-6存在下可诱导的潜伏的HIV基因组 以及其他一些淋巴因子。各种化学致癌物的作用 环境诱变剂检测了它们诱发病毒的能力。 目的是从这个细胞系中识别出 可能影响艾滋病疾病进展的生理学相关性。U1 细胞用2种不同浓度的不同浓度的 致癌物和感染后不同时间的无细胞培养上清液 收集用于测量p24和rt水平的细胞，以及用于 DNA和RNA的制备。用TPA处理细胞作为阳性对照 用于病毒诱导。病毒产量，通过p24抗原检测， 最早在TPA处理的细胞上清液中观察到 时间点，3天。有趣的是，两种被检查的试剂，苯并[2] (BP)和α-六氯己烷(AHCH)诱导的病毒产量相当 与TPA相同，尽管是在较晚的时间点(治疗6-12天)。 在BP和aHCH处理的培养物中观察到RNA水平的增加。一个 通过传播观察到病毒产量增加了10倍 电子显微镜。分析的其他环境致突变物 包括二甲基亚硝胺(DMNA)、尼古丁的各种异构体(NNN， NNK)和黄曲霉毒素B p24抗原水平随DMNA升高5倍，10- 治疗后5天用黄曲霉毒素B折叠，用NNK折叠100倍。 在分析的各种滥用药物中，可卡因、海洛因、LSD和五氯苯酚 病毒抗原诱导倍数为7-20倍。病毒抗原水平显示 黄曲霉毒素B激素诱导的细胞数增加10倍 在这个细胞系统中也在评估病毒的数量。我们目前正在 探讨病毒诱导的机制，RNA和病毒的模式 诱导U1细胞的蛋白质合成以及细胞因子的作用 以及可能控制病毒基因组激活的病毒因子。

The U1 cell line, derived from a chronically infected U937 culture harbors a latent HIV genome which can be induced in the presence of TPA, TNFa, IL-6 and a few other lymphokines. The effect of various chemical carcinogens and environmental mutagens were examined for their ability to induce virus from this cell line with the objective of identifying risk factors of physiologic relevance that may affect disease progression in AIDS. U1 cells were treated with 2 different concentrations of the various carcinogens and at different times after infection, cell-free supernatant was collected for measurement of p24 and RT levels, and cell pellets for DNA and RNA preparation. Cells were treated with TPA as a positive control for viral induction. Virus production, as measured by p24 antigen assays, was observed in the supernatants of cells treated with TPA at the earliest time point, 3 days. Interestingly, 2 of the agents examined, benzopyrene (BP) and alpha hexachlorohexane (aHCH) induced virus production comparable to that of TPA, although at a later time point (6-12 days of treatment). Enhanced levels of RNA were observed in BP and aHCH treated cultures. A ten fold increase in virus production was observed using transmission electron microscopy. Other environmental mutagens that were analyzed included dimethyl nitrosamines (DMNA), various isomers of nicotine (NNN, NNK) and aflatoxin B. p24 antigen levels increased 5 fold with DMNA, 10- fold with aflatoxin B and 100 fold with NNK at five days post treatment. Among the various drugs of abuse analyzed, cocaine, heroine, LSD and PCP showed 7-20 fold induction of viral antigen. Viral antigen levels showed a 10 fold increase in cells treated with Aflatoxin B. Hormonal induction of virus is also being assessed in this cell system. We are currently investigating the mechanism of virus induction, the pattern of RNA and protein synthesis in the induced U1 cells, as well as the role of cellular and viral factors that may control activation of the viral genome.