Bird Sara Whitney, Maynard Nathaniel D, Covert Markus W, Kirkegaard Karla
Departments of Microbiology and Immunology and.
Bioengineering, Stanford University School of Medicine, Stanford, CA 94305.
Proc Natl Acad Sci U S A. 2014 Sep 9;111(36):13081-6. doi: 10.1073/pnas.1401437111. Epub 2014 Aug 25.
The cell-to-cell spread of cytoplasmic constituents such as nonenveloped viruses and aggregated proteins is usually thought to require cell lysis. However, mechanisms of unconventional secretion have been described that bypass the secretory pathway for the extracellular delivery of cytoplasmic molecules. Components of the autophagy pathway, an intracellular recycling process, have been shown to play a role in the unconventional secretion of cytoplasmic signaling proteins. Poliovirus is a lytic virus, although a few examples of apparently nonlytic spread have been documented. Real demonstration of nonlytic spread for poliovirus or any other cytoplasmic constituent thought to exit cells via unconventional secretion requires demonstration that a small amount of cell lysis in the cellular population is not responsible for the release of cytosolic material. Here, we use quantitative time-lapse microscopy to show the spread of infectious cytoplasmic material between cells in the absence of lysis. siRNA-mediated depletion of autophagy protein LC3 reduced nonlytic intercellular viral transfer. Conversely, pharmacological stimulation of the autophagy pathway caused more rapid viral spread in tissue culture and greater pathogenicity in mice. Thus, the unconventional secretion of infectious material in the absence of cell lysis is enabled by components of the autophagy pathway. It is likely that other nonenveloped viruses also use this pathway for nonlytic intercellular spread to affect pathogenesis in infected hosts.
诸如无包膜病毒和聚集蛋白等细胞质成分在细胞间的传播通常被认为需要细胞裂解。然而,已经描述了非常规分泌机制,该机制绕过分泌途径用于细胞质分子的细胞外递送。自噬途径(一种细胞内循环过程)的成分已被证明在细胞质信号蛋白的非常规分泌中起作用。脊髓灰质炎病毒是一种裂解性病毒,尽管已经记录了一些明显非裂解性传播的例子。要真正证明脊髓灰质炎病毒或任何其他被认为通过非常规分泌离开细胞的细胞质成分的非裂解性传播,需要证明细胞群体中的少量细胞裂解与胞质物质的释放无关。在这里,我们使用定量延时显微镜来展示在没有裂解的情况下感染性细胞质物质在细胞间的传播。siRNA介导的自噬蛋白LC3缺失减少了非裂解性细胞间病毒转移。相反,自噬途径的药理学刺激导致病毒在组织培养中传播更快,在小鼠中致病性更强。因此,自噬途径的成分使得在没有细胞裂解的情况下感染性物质能够进行非常规分泌。很可能其他无包膜病毒也利用这一途径进行非裂解性细胞间传播,从而影响感染宿主的发病机制。