The importance of nef in the induction of human immunodeficiency virus type 1 replication from primary quiescent CD4 lymphocytes.
The importance of nef in the induction of human immunodeficiency virus type 1 replication from primary quiescent CD4 lymphocytes.
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DOI：
10.1084/jem.179.1.115
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发表时间：
1994-01-01
期刊：
影响因子：
--
通讯作者：
Richman DD
中科院分区：
文献类型：
--
作者：
Spina CA;Kwoh TJ;Chowers MY;Guatelli JC;Richman DD
病毒调控基因nef是人类免疫缺陷病毒(HIV)及其相关灵长类慢病毒所特有的。在猴免疫缺陷病毒(SIV)感染的动物模型中，nef基因的表达对于维持病毒的高水平复制和发病机制是必不可少的。与这种体内模型相比，使用标准的T细胞培养系统在体外研究nef功能已经产生了一系列相互矛盾的结果，并且未能证明nef对病毒生命周期具有显著的积极影响。我们已经开发了一个细胞模型来研究HIV-1复制的调控，我们认为它更准确地反映了病毒与细胞之间的相互作用，因为它们发生在体内。我们的实验系统使用急性病毒感染纯化的静止的CD4淋巴细胞，然后通过T细胞激活诱导病毒复制。利用该细胞模型，比较了具有开放和突变nef阅读框的NL4-3病毒克隆的复制能力。带有nef突变的克隆在生长速度和最大滴度方面都显示出可重复性和显著的降低。复制减少的程度取决于最初的病毒接种和T细胞激活的时间。在静息状态下，NEF对病毒潜伏状态的病毒复制的诱导作用非常显著。对于完全增殖的CD4细胞的病毒感染，其作用不那么明显。这项研究表明，NEF赋予HIV-1一个积极的生长优势，在通过T细胞激活诱导病毒的初级细胞环境中很容易辨别出来。我们所描述的实验细胞模型不仅为体外研究nef功能提供了手段，也为体内nef在HIV感染中的作用提供了重要线索。
The viral regulatory gene, nef, is unique to the human immunodeficiency viruses (HIV) and their related primate lentiviruses. Expression of the nef gene has been shown to be essential to the maintenance of high levels of virus replication and the development of pathogenesis in the animal model of simian immunodeficiency virus (SIV) infection. In contrast to this in vivo model, the use of standard T cell culture systems to study nef function in vitro has produced a spectrum of contradictory results, and has failed to demonstrate a significant positive influence of nef on viral life cycle. We have developed a cell model to study regulation of HIV-1 replication that we believe reflects more accurately virus-cell interactions as they occur in vivo. Our experimental system used acute virus infection of purified, quiescent CD4 lymphocytes and subsequent induction of viral replication through T cell activation. With this cell model, NL4-3 virus clones with open and mutated nef reading frames were compared for replication competence. The clones with nef mutations showed reproducible and significant reductions in both rates of growth and maximal titers achieved. The degree of reduced replication was dependent on initial virus inoculum and the timing of T cell activation. The influence of nef was highly significant for induction of virus replication from a latent state within resting CD4 cells. Its effect was less apparent for virus infection of fully proliferating CD4 cells. This study demonstrates that nef confers a positive growth advantage to HIV-1 that becomes readily discernable in the primary cell setting of virus induction through T cell activation. The experimental cell model, which we describe here, provides not only a means to study nef function in vitro, but also provides important clues to the function of nef in HIV infection in vivo.