A dominant block to HIV-1 replication at reverse transcription in simian cells
A dominant block to HIV-1 replication at reverse transcription in simian cells
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DOI：
10.1073/pnas.212400099
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发表时间：
2002-10-15
影响因子：
11.1
通讯作者：
Landau, NR
中科院分区：
文献类型：
--
作者：
Münk, C;Brandt, SM;Landau, NR
尽管非人类灵长类动物在基因上与人类接近，但它们的 T 细胞不支持 HIV-1 的高效复制。相比之下，HIV-1 在激活的人类 CD4+ T 细胞、单核细胞和多种细胞类型的代谢活跃的人类细胞中复制，当转导表达 CD4 和 CCR5 或 CXCR4 时，它们变得允许 HIV-1 复制。通过使用稳定转导表达人类 CD4 和 CCR5 的非洲绿猴和恒河猴细胞系，研究了该物种限制 HIV-1 复制的分子基础。这些细胞支持同源病毒[来自非洲绿猴（SIV-AGM）和猕猴（SIVmac239）的猿猴免疫缺陷病毒]的复制，但不支持R5亲性细胞病变HIV-1的复制。基于 β-内酰胺酶的 HIV-1 进入测定表明该病毒有效地进入非人类灵长类动物细胞。与类似感染的人类细胞相比，原病毒的形成减少了 50 倍。实时 PCR 定量表明，猴细胞中的逆转录未能有效启动。当感染复数 >1 时，或通过将非人灵长类细胞与病毒预孵育，逆转录的阻断被覆盖，这一特征让人想起 Friend 病毒抗性基因 1 (FV-1)，限制小鼠白血病病毒在小鼠细胞中的复制。人类和猿细胞融合的异核分析表明，块是占主导地位的。这些发现表明灵长类动物细胞含有一种主要抑制剂，可以阻止 HIV-1 逆转录。
Although nonhuman primates are genetically close to humans, their T cells do not support productive replication of HIV-1. In contrast, HIV-1 replicates in activated human CD4+ T cells, monocytes, and metabolically active human cells of a variety of cell types become permissive for HIV-1 replication when transduced to express CD4 and CCR5 or CXCR4. The molecular basis of this species restriction to HIV-1 replication was investigated by using African green monkey and rhesus macaque cell lines that were stably transduced to express human CD4 and CCR5. The cells supported replication of cognate viruses [simian immunodeficiency virus from African green monkeys (SIV-AGM) and macaques (SIVmac239)] but did not support replication of an R5-tropic cytopathic HIV-1. A beta-lactamase-based HIV-1 entry assay was used to show that the virus efficiently entered the nonhuman primate cells. Provirus formation was reduced 50-fold compared with similarly infected human cells. Real-time PCR quantitation demonstrated that reverse transcription failed to initiate efficiently in the simian cells. The block to reverse transcription was overridden at multiplicity of infection >1 or by preincubation of the nonhuman primate cells with virus, a feature reminiscent of the Friend virus resistance gene-1 (FV-1), restriction to murine leukemia virus replication in mouse cells. Heterokaryon analysis in which human and simian cells were fused demonstrated that the block was dominant. These findings suggested that the primate cells contain a dominant inhibitor that prevents HIV-1 reverse transcription.