Vela Eric M, Zhang Lihong, Colpitts Tonya M, Davey Robert A, Aronson Judith F
Department of Pathology, Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555, USA.
Virology. 2007 Dec 5;369(1):1-11. doi: 10.1016/j.virol.2007.07.014. Epub 2007 Aug 14.
Arenaviruses are important causes of viral hemorrhagic fevers in humans. Arenavirus infection of cells occurs via a pH-dependent endocytic route, but detailed studies of entry pathways have not been done. We investigated the role of cell membrane cholesterol, caveolae, and clathrin coated pits in infection by Lassa virus (LASV), which utilizes alpha-dystroglycan (alpha-DG) as a receptor, and Pichindé virus (PICV), which does not. Depletion of cellular cholesterol by treatment with methyl betacyclodextrin (MbetaCD) or nystatin/progesterone inhibited PICV replication and transfer of packaged marker gene by LASV or PICV pseudotyped retroviral particles. In cells lacking caveolae due to silencing of the caveolin-1 gene, no inhibition of PICV infection or LASV pseudotype transduction was observed. However, PICV infection and LASV and PICV pseudotype transduction was inhibited when an Eps15 dominant negative mutant was used to inhibit clathrin-mediated endocytosis. Altogether, the results indicate that diverse arenaviruses have a common requirement for cell membrane cholesterol and clathrin mediated endocytosis in establishing infection.
沙粒病毒是人类病毒性出血热的重要病因。沙粒病毒感染细胞是通过pH依赖的内吞途径发生的，但尚未对其进入途径进行详细研究。我们研究了细胞膜胆固醇、小窝和网格蛋白包被小窝在拉沙病毒（LASV）感染中的作用，拉沙病毒利用α- dystroglycan（α-DG）作为受体；同时也研究了皮钦德病毒（PICV）感染中的作用，皮钦德病毒不利用α-DG作为受体。用甲基β-环糊精（MbetaCD）或制霉菌素/孕酮处理使细胞胆固醇耗竭，可抑制PICV复制以及LASV或PICV假型逆转录病毒颗粒包装的标记基因的转移。在因小窝蛋白-1基因沉默而缺乏小窝的细胞中，未观察到对PICV感染或LASV假型转导的抑制作用。然而，当使用Eps15显性负性突变体抑制网格蛋白介导的内吞作用时，PICV感染以及LASV和PICV假型转导均受到抑制。总之，结果表明，不同的沙粒病毒在建立感染过程中对细胞膜胆固醇和网格蛋白介导的内吞作用有共同需求。
