VARICELLA-ZOSTER--RECEPTORS AND INFECTIVE MECHANISMS

水痘带状疱疹——受体和感染机制

• 批准号: 3141336
• 负责人: Anne A. Gershon
• 金额: $ 14.38万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-12-01 至 1991-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3141336
• 关键词: Betaherpesvirinae; Herpes simplex disease; Herpesviridae; Vesiculovirus; affinity chromatography; aniline dye; asparagine; autoradiography; beta glucuronidase; biological signal transduction; biomarker; cell membrane; chloroquine; conformation; enzyme linked immunosorbent assay; exocytosis; fibroblasts; fluorescent dye /probe; glucosamine; glycoprotein biosynthesis; glycoproteins; glycosylation; human tissue; hydrolase; immunocytochemistry; immunoelectron microscopy; mannose 6 phosphate isomerase; membrane permeability; monoclonal antibody; monosaccharides; nuclear membrane; oligosaccharides; radiotracer; skin; thymidine monophosphate; tritium; varicella zoster virus; virus envelope; virus infection mechanism; virus protein; virus receptors; virus replication

The intracellular assembly, maturation, and transport of varicella-zoster virus (VZV) will be studied. VZV, the highly contagious etiologic agent of chickenpox (varicella) and shingles (zoster) is spread by infectious virions that are present in fluid that fills vesicular skin lesions. Virions secreted by cells in vitro, however, are not infectious and yield infectious particles only when the cells are disrupted. EM observations suggest that nucleocapsids of VZV are enveloped while exiting from the nucleus through the inner nuclear membrane. Virions then travel within the cisternal compartment of the Golgi apparatus where they are diverted from the secretory pathway to prelysosomal structures. Diverted virions accumulate in acid phosphatase-containing vacuoles in which the particles appear to be degraded. These observations are consistent with the hypothesis that the intracellular route of transport of newly synthesized VZV particles is similar to that of newly synthesized acid hydrolases. We have found that VZV envelope glycoproteins (gps), like the acid hydrolases, contain mannose 6-phosphate (Man 6-P) residues. We postulate that Man 6-P on the viral envelope leads VZV to bind to Man 6-P receptors (MPRs) in the Golgi, which, in turn, direct newly synthesized VZV to endosomes. MPRs also cycle to the cell surface; therefore, if VZV were to escape degradation in the epidermis of infected patients, then Man 6-P on the viral envelope might bind to MPRs on the plasma membrane of target cells, facilitation infection. We will test the hypotheses that VZV binds to MPRs, that intracellular binding of VZV to MPRs in cultured cells diverts virions from the secretory to the lysosomal pathway, that binding of intact VZV to surface MPRs facilities viral infection of target cells, and that cells in suprabasal layers of the epidermis of infected patients lack intracellular MPRs and thus secrete infectious virions into vesicular fluid. Preliminary data indicate that: (i) Man 6-P is present on complex oligosaccharides of viral envelope gps. (ii) phosphorylated monosaccharides protect cells from infection by cell-free VZV in vitro; (iii) the rank order of efficacy (Man 6-P>>Glu 6-P>Glu 1-P) is the same as that for affinity for MPRs; (iv) chloroquine, which prevents recycling of free MPRs to the cell surface, reversibly protects cells from infection by VZV; (v) VZV accumulates in endosomes and the trans Golgi network of infected cells in culture and VZV gp immunoreactivity is co-distributed in cells with that of MPRs. We propose to definitively identify the compartments of the intracellular pathway of viral maturation. We will ascertain where VZV gps become incorporated into the viral envelope. We will determine the structures of VZV-associated oligosaccharides and how they are synthesized. We will establish if VZV-associated Man 6-P bearing oligosaccharides are sufficient to mediate binding of VZV to MPRs, we will assay the presence of Man 6-P on intact VZV virions obtained from vesicular fluid of cutaneous lesions, we will test the role of MPRs in mediating VZV infection of cultured cells, and we will investigate how VZV escapes intracellular degradation and reaches vesicular fluid in cutaneous lesions of patients with varicella.

水痘-带状疱疹的细胞内组装、成熟和转运 病毒（VZV）的研究。 VZV是一种高度传染性的病原体， 水痘（水痘）和带状疱疹（带状疱疹）是通过传染性传播的 存在于充满水疱性皮肤损伤的液体中的病毒体。 然而，由细胞在体外分泌的病毒粒子不具有感染性， 只有当细胞被破坏时才有感染性颗粒。 EM观察 表明VZV的核衣壳在离开细胞时被包裹， 通过内核膜进入细胞核。 然后病毒粒子在 高尔基体的池状隔室，它们从 前溶酶体结构的分泌途径。 转向病毒粒子 在含有酸性磷酸酶的液泡中积累， 似乎被降级了。 这些观察结果与 假设新合成的细胞内转运途径 VZV颗粒类似于新合成的酸性水解酶。 我们 已经发现VZV包膜糖蛋白（GPS），像酸性水解酶， 含有甘露糖6-磷酸（Man 6-P）残基。 我们假设Man 6-P 病毒包膜上的VZV导致VZV结合到细胞膜上的Man 6-P受体（MPR）。 高尔基体，而高尔基体又将新合成的VZV引导至内体。 MPRS 也循环到细胞表面;因此，如果VZV逃脱， 在感染患者的表皮降解，然后曼6-P对 病毒包膜可能与靶细胞质膜上的MPR结合， 易化感染 我们将测试VZV与 MPRs，即VZV与培养细胞中MPRs的细胞内结合转移 从分泌途径到溶酶体途径的病毒体， VZV与表面MPR的结合促进了靶细胞的病毒感染， 感染患者表皮的基底上层细胞缺乏 从而将感染性病毒体分泌到囊泡中 液 初步数据表明：（i）Man 6-P存在于复合物上， 病毒包膜GPS的低聚糖。 (ii)磷酸化 单糖在体外保护细胞免受无细胞VZV的感染; (iii)功效的等级顺序（Man 6-P>> Glu 6-P> Glu 1-P）与 （iv）氯喹，其防止MPRs的再循环。 游离MPR到细胞表面，可逆地保护细胞免受感染， VZV;（v）VZV在核内体和高尔基体网络中积累。 感染的细胞培养和VZV gp免疫反应性共分布在 细胞与MPR的细胞。 我们建议明确确定 病毒成熟的细胞内途径的区室。 我们将 确定VZV GPS在哪里被并入病毒包膜。 我们 将确定VZV相关寡糖的结构以及如何 它们是合成的。 我们将确定VZV相关的Man 6-P方位 寡糖足以介导VZV与MPR的结合，我们将 测定从囊泡中获得的完整VZV病毒体上Man 6-P的存在 我们将测试MPR在介导VZV中的作用， 感染培养细胞，我们将研究VZV如何逃脱 细胞内降解并到达皮肤病变的囊液 水痘的患者。