Stimulated trans-acting factor of 50 kDa (Staf50) inhibits HIV-1 replication in human monocyte-derived macrophages
Stimulated trans-acting factor of 50 kDa (Staf50) inhibits HIV-1 replication in human monocyte-derived macrophages
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DOI：
10.1016/j.virol.2006.07.025
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发表时间：
2006-12-01
期刊：
影响因子：
3.7
通讯作者：
von Briesen, Hagen
中科院分区：
文献类型：
--
作者：
Bouazzaoui, Abdellatif;Kreutz, Marina;von Briesen, Hagen
为了鉴定干扰单核细胞衍生的巨噬细胞（MAC）中HIV-1复制的细胞基因，用干扰素（IFN）或脂多糖（LPS）刺激细胞，导致这些细胞中HIV-1感染的显著抑制，并分析所得基因表达。使用微阵列技术，我们鉴定了一个名为50 kDa的刺激反式作用因子（Staf 50）的基因，该基因已知抑制HIV-1 LTR的活性。通过实时PCR分析Staf 50表达显示在IFN α（高达20倍）和LPS（高达10倍）刺激的MAC中以及在感染的细胞中过表达（高达3倍）。为了稳定过表达，用Staf 50-IRES-GFP双顺反子假型病毒转导293个T细胞和原代巨噬细胞。转导后，用HIV-1感染293 T CD 4/CCR 5和MAC，并用p24 ELISA监测病毒复制。Staf 50的过表达对293 T CD 4/CCR 5细胞和MAC细胞的HIV-1感染抑制率在50%~ 90%之间。我们的研究结果表明，宿主遗传效应与病毒特性相结合，决定了适当的靶细胞对HIV-1感染的易感性，以及病毒在细胞中的复制潜力，从而导致整体的生产性感染。(c)2006年爱思唯尔公司All rights reserved.
In order to identify cellular genes which interfere with HIV-1 replication in monocyte-derived macrophages (MAC), cells were stimulated with interferon (IFN) or lipopolysaccharide (LPS) leading to a pronounced inhibition of HIV-1 infection in these cells, and the resulting gene expression was analyzed. Using the microarray technology we identified a gene named Stimulated Trans-Acting Factor of 50 kDa (Staf50), which is known to repress the activity of the HIV-1 LTR. Analysis of the Staf50 expression by real-time PCR showed an overexpression in IFN alpha (up to 20-fold) and LPS (up to 10-fold)-stimulated MAC as well as in infected cells (up to 3-fold). For stable overexpression, 293 T cells and primary macrophages were transduced with Staf50-IRES-GFP bicistronic pseudotype viruses. After transduction, 293 T CD4/CCR5 and MAC were infected with HIV-1, and virus replication was monitored by p24 ELISA. Overexpression of Staf50 inhibited the HIV-1 infection between 50% and 90% in 293 T CD4/CCR5 as well as in MAC. Our findings suggest that host genetic effects in combination with viral properties determine the susceptibility of an appropriate target cell for HIV-1 infection as well as the replication potential of the virus in the cell resulting in an overall productive infection. (c) 2006 Elsevier Inc. All rights reserved.