Myosin-X is essential to the intercellular spread of HIV-1 Nef through tunneling nanotubes.
Myosin-X is essential to the intercellular spread of HIV-1 Nef through tunneling nanotubes.
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肌球蛋白-X 对于 HIV-1 Nef 通过隧道纳米管在细胞间传播至关重要。
DOI：
10.1007/s12079-018-0493-z
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发表时间：
2019
影响因子：
4.1
通讯作者：
Gousset,Karine
中科院分区：
文献类型：
--
作者：
Uhl,Jaime;Gujarathi,Shivalee;Waheed,AbdulA;Gordon,Ana;Freed,EricO;Gousset,Karine
纳米管是一种细胞间结构，可供囊泡、细胞器、基因组物质、致病蛋白和病原体通过。非常规肌动蛋白分子马达蛋白Myosin-X（Myo 10）是神经元细胞中TNTs的已知诱导剂，但其在其他细胞类型中的作用尚未被研究。Nef HIV-1辅助蛋白对于HIV-1发病机制至关重要，并且可以通过TNT在培养物中自我传播。了解它的细胞间传播机制可以揭示在HIV-1感染期间控制其破坏作用的方法。本研究的目的是探讨Nef从巨噬细胞到T细胞的细胞间转运机制。我们证明，Nef在巨噬细胞中以Myo 10依赖的方式增加TNT，并观察到Nef通过TNT从巨噬细胞转移到T细胞。为了定量这种转移机制，我们建立了间接流式细胞术测定。由于Nef在T细胞中的表达下调表面受体CD 4，我们将CD 4的减少与Nef在这些细胞之间的转移相关联。因此，我们将表达不同水平Nef的巨噬细胞与表达高水平CD 4的T细胞系共培养，并定量由Nef转移引起的CD 4表面表达的变化。我们证明，Nef转移发生通过细胞到细胞的依赖性机制，直接与Myo 10依赖性TNT的存在。因此，我们表明Nef可以调节Myo 10的表达，从而诱导TNT的形成，导致其自身从巨噬细胞转移到T细胞。此外，我们证明，上调Myo 10诱导Nef也发生在人类单核细胞衍生的巨噬细胞在HIV-1感染。
Tunneling nanotubes (TNTs) are intercellular structures that allow for the passage of vesicles, organelles, genomic material, pathogenic proteins and pathogens. The unconventional actin molecular motor protein Myosin-X (Myo10) is a known inducer of TNTs in neuronal cells, yet its role in other cell types has not been examined. The Nef HIV-1 accessory protein is critical for HIV-1 pathogenesis and can self-disseminate in culture via TNTs. Understanding its intercellular spreading mechanism could reveal ways to control its damaging effects during HIV-1 infection. Our goal in this study was to characterize the intercellular transport mechanism of Nef from macrophages to T cells. We demonstrate that Nef increases TNTs in a Myo10-dependent manner in macrophages and observed the transfer of Nef via TNTs from macrophages to T cells. To quantify this transfer mechanism, we established an indirect flow cytometry assay. Since Nef expression in T cells down-regulates the surface receptor CD4, we correlated the decrease in CD4 to the transfer of Nef between these cells. Thus, we co-cultured macrophages expressing varying levels of Nef with a T cell line expressing high levels of CD4 and quantified the changes in CD4 surface expression resulting from Nef transfer. We demonstrate that Nef transfer occurs via a cell-to-cell dependent mechanism that directly correlates with the presence of Myo10-dependent TNTs. Thus, we show that Nef can regulate Myo10 expression, thereby inducing TNT formation, resulting in its own transfer from macrophages to T cells. In addition, we demonstrate that up-regulation of Myo10 induced by Nef also occurs in human monocyte derived macrophages during HIV-1 infection.