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

单纯疱疹病毒I型（HSV 1）在粘膜中初始复制后在周围神经节中潜伏感染。HSV 1很少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。HSV 1衣壳在细胞核中组装。它们通过在内核膜上发芽获得初级被膜，内核膜通过与外核膜融合而脱落。胞质衣壳然后被运输到膜细胞器，在那里它们通过出芽获得次级包膜。产生的感染性病毒体通过分泌或细胞死亡释放。我们将研究神经元细胞培养模型中HSV 1的继发性增殖，以及它与感染的轴突扩散的关系。使用（1）荧光和电子显微镜中的特异性抗体，（2）在同步HSV 1感染期间同时体内标记病毒结构和细胞出芽区室的数字延时视频显微镜，和（3）我们将在我们的HSV 1-BAC中产生的内层蛋白VP 1/3（UL 36）和UL 37的病毒突变体，我们将确定病毒和宿主因子，这些因子对神经元中HSV 1的次级包膜至关重要。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。介导微管转运和次级介导的病毒和宿主蛋白质可为开发新的抗HSV 1和其他疱疹病毒的抗病毒化疗提供特异性靶结构。

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.