The dual role of tetraspanin CD63 in HIV-1 replication.
The dual role of tetraspanin CD63 in HIV-1 replication.
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四跨果蛋白CD63在HIV-1复制中的双重作用。
DOI：
10.1186/1743-422x-11-23
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发表时间：
2014-02-08
期刊：
影响因子：
4.8
通讯作者：
Ferguson MR
中科院分区：
文献类型：
--
作者：
Li G;Endsley MA;Somasunderam A;Gbota SL;Mbaka MI;Murray JL;Ferguson MR
以前，我们表明，四跨膜蛋白CD 63介导的HIV-1复制周期的早期和后期整合阶段。使用单克隆抗体和小干扰RNA（siRNA）来阻断CD 63功能，并确定HIV-1复制中哪些顺序步骤被破坏，从而识别CD 63的时间作用。在早期感染过程中出现抑制，提示参与病毒进入或逆转录。此外，我们已经表明，感染后用CD 63 siRNA处理，显著抑制上清液中的病毒产生，表明CD 63在前病毒整合后发生的HIV-1复制事件中以及可能在排出过程中在巨噬细胞中起重要作用。在这项研究中，我们使用CD 63 siRNA研究假型病毒（携带NL 4 -3 Env阴性荧光素酶骨架）在原代人巨噬细胞中的感染性。我们证明了实验室适应的R5和R5 X4嗜性HIV-1毒株被CD 63沉默显著抑制。然而，通过受体介导的内吞作用进入的MLV或VSV假型毒株的感染性不受沉默CD 63的影响。这些结果表明，CD 63可以支持通过CD 4和CCR 5促进的Env介导的进入或融合事件。此外，基于抗体和siRNA的CD 63抑制研究表明CD 63在前病毒整合后的潜在作用。此外，我们表明，CD 63的表达是在原代CD 4 + T细胞中有效复制的关键，补充了我们先前对原代人巨噬细胞和永生化细胞系的研究。总的来说，这些发现表明，CD 63可能支持Env介导的融合以及HIV-1复制周期中的后期（整合后）步骤。
Previously, we showed that the tetraspanin membrane protein CD63 mediates both early and post-integration stages of the HIV-1 replication cycle. The temporal roles of CD63 were discerned using monoclonal antibodies and small interfering RNAs (siRNAs) to block CD63 function, and determining which of the sequential steps in HIV-1 replication were disrupted. Inhibition was shown to occur during early infection, suggestive of involvement in virus entry or reverse transcription. In addition, we have shown that treatment with CD63 siRNA post-infection, significantly inhibited virus production in supernatant, suggesting an important role for CD63 in macrophages during HIV-1 replication events occurring after proviral integration, and possibly during egress. In this study we used CD63 siRNA to investigate the infectivity of pseudotyped viruses (carrying an NL4-3 Env-negative luciferase backbone) in primary human macrophages. We demonstrated that lab adapted R5- and R5X4-tropic HIV-1 strains are significantly inhibited by CD63 silencing. However, the infectivity of MLV or VSV-pseudotyped strains, which enter though receptor-mediated endocytosis, is unaffected by silencing CD63. These results indicate that CD63 may support Env-mediated entry or fusion events facilitated though CD4 and CCR5. Also, antibody and siRNA-based CD63 inhibition studies indicate a potential role for CD63 following proviral integration. Further, we show that CD63 expression is key for efficient replication in primary CD4+ T cells, complementing our prior studies with primary human macrophages and immortalized cell lines. Collectively, these findings indicate that CD63 may support Env-mediated fusion as well as a late (post-integration) step in the HIV-1 replication cycle.