DISTINCT MODES OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PROVIRAL LATENCY REVEALED BY SUPERINFECTION OF NONPRODUCTIVELY INFECTED CELL-LINES WITH RECOMBINANT LUCIFERASE-ENCODING VIRUSES
DISTINCT MODES OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PROVIRAL LATENCY REVEALED BY SUPERINFECTION OF NONPRODUCTIVELY INFECTED CELL-LINES WITH RECOMBINANT LUCIFERASE-ENCODING VIRUSES
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DOI：
10.1128/jvi.68.2.654-660.1994
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发表时间：
1994-02-01
影响因子：
5.4
通讯作者：
BALTIMORE, D
中科院分区：
文献类型：
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作者：
CHEN, BK;SAKSELA, K;BALTIMORE, D
为了研究人类免疫缺陷病毒（HIV）的细胞潜伏期的基础，我们使用了一种编码HIV的重组蛋白酶（HXB Luc）来超感染非生产性HIV-1感染的人白血病细胞系。HXB-Luc含有Photinus pyralis荧光素酶基因代替nef基因，并为HIV感染和表达提供了高度灵敏、简单的检测方法。为了规避潜伏感染细胞中的任何重复感染阻断，我们还产生了用鼠白血病病毒嗜中性包膜（HXB-Lucampho）假型化的病毒。亲本未感染细胞系U937和A3.01分别衍生出潜伏感染的细胞系U1和ACH-2，它们可以容易地被假型或非假型报告病毒感染。然而，用HXB-Luc或HXB-Lucampho重复感染U1细胞仅导致低水平的荧光素酶活性。与内源性前病毒一样，HXB-Luc前病毒可以被HXB-Lucampho超感染的U1细胞的佛波醇酯处理有效地激活。相比之下，ACH-3细胞的重复感染甚至在未刺激的细胞中也导致二次引入的病毒活跃表达，并且荧光素酶产量高于亲代细胞系A3.01。因此，U1细胞中的前病毒潜伏期似乎是由细胞环境中的缺陷引起的（反式效应），而ACH-2中的潜伏期对整合的前病毒是特异性的，并且可能是由于整合位点的顺式效应。这些结果证明了这两种细胞系模型中前病毒潜伏期的不同模式，并可能对我们理解HIV感染中细胞潜伏期的调节和意义产生影响。
To study the basis of cellular latency of human immunodeficiency virus (HIV), we have used a recombinant luciferase-encoding HIV (HXB-Luc) to superinfect nonproductively HIV-1-infected human leukemic cell lines. HXB-Luc contains the Photinus pyralis luciferase gene in place of the nef gene and provides a highly sensitive, simple assay for HIV infection and expression. To circumvent any superinfection block in latently infected cells, we also generated viruses pseudotyped with murine leukemia virus amphotropic envelope (HXB-Lucampho). The parental uninfected lines, U937 and A3.01, from which the latently infected cell lines U1 and ACH-2, respectively, were derived could he readily infected with pseudotyped or nonpseudotyped reporter viruses. However, superinfection of U1 cells with either HXB-Luc or HXB-Lucampho resulted in only low levels of luciferase activity. Like the endogenous provirus, HXB-Luc provirus could be efficiently activated by phorbol ester treatment of HXB-Lucampho-superinfected U1 cells. In contrast, superinfection of ACH-3 cells resulted in active expression of the secondarily introduced virus even in unstimulated cells and luciferase production higher than in the parental cell line A3.01. Thus, the proviral latency in U1 cells appears to result from a defect in the cellular environment (a trans effect), whereas the latency in ACH-2 is specific to the integrated provirus and is probably a cis effect due to the site of integration. These results demonstrate distinct modes of proviral latency in these two cell Line models and may have implications in our understanding of the regulation and significance of cellular latency in HIV infection.