Human immunodeficiency virus type 1 replication is blocked prior to reverse transcription and integration in freshly isolated peripheral blood monocytes
Human immunodeficiency virus type 1 replication is blocked prior to reverse transcription and integration in freshly isolated peripheral blood monocytes
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DOI：
10.1128/jvi.70.6.3863-3869.1996
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发表时间：
1996-06-01
影响因子：
5.4
通讯作者：
Crowe, S
中科院分区：
文献类型：
--
作者：
Sonza, S;Maerz, A;Crowe, S
外周血单核细胞在分离后立即在体外对生产性人类免疫缺陷病毒1型（HIV-1）感染具有抵抗力，在分离当天暴露于病毒的单核细胞中通过PCR检测不到病毒cDNA（无论是早期还是晚期转录物）。相比之下，在单核细胞培养短短1天，启动和完成逆转录病毒感染HIV-1（BA-L）和原代分离株的24小时内很容易检测。感染后24小时发现的启动，部分完成和完成的病毒DNA拷贝的水平随着感染前培养时间的推移而逐渐增加。与静止的T淋巴细胞不同，似乎没有阻断或延迟病毒DNA整合到易感培养的单核细胞的基因组中。使用设计用于特异性检测前病毒DNA的ALL-PCR方法，在分离后培养一天或更长时间的单核细胞中，在感染后24小时内发现整合事件。在暴露于病毒后7天内，在新鲜分离的单核细胞中未发现整合信号。经RT-PCR扩增的DNA的克隆和测序证实整合在HIV-1感染的培养单核细胞中。我们的研究结果表明，在体外复制的HIV-1在新鲜分离的外周血单核细胞的逆转录启动之前，显然是封闭的，这表明这些细胞可能不容易感染体内。进一步的研究，以澄清这种可能性和性质的块感染应提供有用的信息，治疗策略对HIV-1。
Peripheral blood monocytes are resistant to productive human immunodeficiency virus type 1 (HIV-1) infection in vitro immediately after isolation, No viral cDNA (either early or late transcripts) was detected by PCR in monocytes exposed to virus on the day of isolation. In contrast, in monocytes cultured for as little as 1 day, initiated and completed reverse transcripts were readily detectable within 24 h of infection with both HIV-1(BA-L) and primary isolates. The levels of initiated, partially completed, and completed viral DNA copies found 24 h after infection increased progressively with time in culture before infection. Unlike quiescent T lymphocytes, there appeared to be no block or delay in the integration of viral DNA into the genome of susceptible cultured monocytes, With an ALL-PCR method designed to specifically detect proviral DNA being used, integration events were found within 24 h of infection in monocytes cultured for a day or more after isolation. No integration signal was found in freshly isolated monocytes up to 7 days following exposure to the virus. Cloning and sequencing of Alu-PCR-amplified DNA confirmed integration in HIV-1-infected cultured monocytes. Our finding that in vitro replication of HIV-1 is clearly blocked prior to the initiation of reverse transcription in freshly isolated peripheral blood monocytes suggests that these cells may not be susceptible to infection in vivo. Further studies to clarify this possibility and the nature of the block to infection should provide useful information for treatment strategies against HIV-1.