Maldarelli F, Sato H, Berthold E, Orenstein J, Martin M A
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892, USA.
J Virol. 1995 Oct;69(10):6457-65. doi: 10.1128/JVI.69.10.6457-6465.1995.
The kinetics of human immunodeficiency virus type 1 (HIV-1)-induced cell death were investigated in cell-to-cell and cell-free models of virus transmission. Cocultivation of HIV-1 chronically infected H9 donor cells with uninfected H9 recipient cells resulted in rapid induction of programmed cell death. Within 8 h, apoptotic chromatin condensation was identified by histologic staining. In addition, many single cells with apoptotic nuclei were observed, indicating that stable cell fusion was not a requirement for apoptosis to occur. By 12 to 18 h of coculture, a DNA fragmentation ladder characteristic of apoptosis was detected by agarose gel electrophoresis. Quantitation of apoptosis by measurement of nuclear DNA content revealed that at least 20 to 30% of the nuclei were undergoing apoptosis by 24 h after cocultivation. The appearance of condensed nuclei and fragmented DNA occurred as HIV reverse transcription was completed, and it was not inhibited by zidovudine, suggesting that induction of apoptosis did not require new HIV replication. Soluble CD4 inhibited apoptosis, demonstrating that Env-CD4 interactions were required for apoptosis. In contrast to that in cell-to-cell transmission, apoptosis in cell-free HIV infections was markedly inefficient and was not observed until 70 to 90 h after infections were initiated. These findings indicate that HIV-1 induction of programmed destruction of the nucleus is initiated at the time of cell-cell cocultivation by a mechanism which requires CD4-Env interactions but not new HIV replication.
在病毒传播的细胞间和无细胞模型中研究了1型人类免疫缺陷病毒(HIV-1)诱导的细胞死亡动力学。将HIV-1慢性感染的H9供体细胞与未感染的H9受体细胞共培养,可快速诱导程序性细胞死亡。8小时内,通过组织学染色可识别出凋亡染色质浓缩。此外,观察到许多具有凋亡细胞核的单个细胞,表明凋亡的发生并不需要稳定的细胞融合。共培养12至18小时后,通过琼脂糖凝胶电泳检测到凋亡特有的DNA片段梯带。通过测量核DNA含量对凋亡进行定量分析表明,共培养24小时后,至少20%至30%的细胞核正在发生凋亡。浓缩细胞核和DNA片段化的出现发生在HIV逆转录完成时,且不受齐多夫定抑制,这表明凋亡的诱导不需要新的HIV复制。可溶性CD4可抑制凋亡,表明Env-CD4相互作用是凋亡所必需的。与细胞间传播不同,无细胞HIV感染中的凋亡明显低效,直到感染开始70至90小时后才观察到。这些发现表明,HIV-1通过一种需要CD4-Env相互作用但不需要新的HIV复制的机制,在细胞间共培养时启动对细胞核的程序性破坏。