The function of microtubules for the secondary envelopment of Herpes Simplex Virus in neurons

单纯疱疹病毒1型（HSV1）在外周神经节粘膜潜伏感染中初始复制后建立。罕见的是，HSV1侵入中枢神经系统并引起疱疹性脑炎，如果不治疗，70%的病例是致命的。HSV1衣壳在细胞核内组装。它们通过与外核膜融合而脱落的内核膜出芽获得初级包膜。细胞质衣壳随后被运送到膜细胞器，在那里它们通过出芽获得二级包膜。由此产生的传染性病毒粒子通过分泌或细胞死亡释放出来。我们将研究HSV1在神经元细胞培养模型中的继发包膜，以及它与感染轴突传播的关系。利用(1)荧光和电子显微镜下的特异性抗体，(2)同步感染HSV1时，同时在体内标记病毒结构和细胞出芽室的数字定时视频显微镜，以及(3)我们将在HSV1- bac中产生的内被膜蛋白VP1/3 （UL36）和UL37的病毒突变体，我们将确定对神经元中HSV1继发包膜发育至关重要的病毒和宿主因素。我们将讨论微管网络在亚病毒颗粒和感染性病毒的神经元内运输中的作用。介导微管转运和二次包膜的病毒和宿主蛋白可为开发新的HSV1和其他疱疹病毒抗病毒化疗药物提供特异性靶点结构。

Herpes Simplex Virus type l (HSV1) establishes after initial replication in the mucosa latent infections in peripheral nervous ganglia. Rarely, HSV1 invades the central nervous system and causes Herpes encephalitis that if untreated is lethal in 70% of all cases. HSV1 capsids assemble in the cell nuclei. They acquire a primary envelope by budding at the inner nuclear membrane that is shed by fusion with the outer nuclear membrane. Cytosolic capsids are then transported to a membrane organelle where they acquire a secondary envelope by budding. The resulting infectious virions are released by secretion or cell death. We will study the secondary envelopment of HSV1 in neuronal cell culture models, and how that is related to axonal spread of an infection. Using (1) specific antibodies in fluorescence and electron microscopy, (2) digital time-lapse video microscopy with simultaneously in vivo labelling of viral structures and the cellular budding compartment during a synchronized HSV1 infection, and (3) viral mutants of the inner tegument proteins VP1/3 (UL36) and UL37 that we will generate in our HSV1-BAC, we will identify the viral and host factors that are crucial for HSV1 secondary enve-lopment in neurons. We will address the role of the microtubule network in the intraneuronal transport of subviral particles and infectious virus. The viral and host proteins that mediate microtubule transport and secondary envelopment could provide specific target structures for the development of new antiviral chemotherapy of HSV1 and other herpesviruses.