Adaptations of Human Herpesvirus-7 to Salivary Glands

人类疱疹病毒 7 号对唾液腺的适应

• 批准号: 6361154
• 负责人: Stephen Dewhurst
• 金额: $ 30.31万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6361154
• 关键词: Alphaherpesvirinae; Betaherpesvirinae; cell cell interaction; cell membrane; chemokine; cytomegalovirus; epithelium; epitope mapping; gene expression; human tissue; laboratory mouse; laboratory rat; membrane fusion; monoclonal antibody; neutralizing antibody; protein structure function; proteoglycan; recombinant virus; salivary glands; tissue /cell culture; virus antigen; virus genetics; virus infection mechanism; virus protein; virus replication

Human herpesvirus (HHV)-7 infection is associated with persistence of viral genomes in salivary gland (SG) tissues, and chronic expression of viral antigens in these sites. The virus is generally thought to be spread by a salivary route, and there is lifelong shedding of large amounts of infectious virions in saliva. These observations suggest the following hypothesis: that HHV-7 has evolved specific mechanisms to gain entry to SG cells and to evade host immune responses in SG tissues. In this proposal, we will examine the role of specific viral proteins in virus attachment and entry to SG epithelial cells. In the first two specific aims, the unique viral glycoprotein, gp65, will be studied. Gp65 is a component of the virus particle, and polyclonal antisera directed against gp65 neutralize virus infectivity; HHV-7 gp65 also binds to heparan sulfate proteoglycans (HSPGs). These data strongly suggest that gp65 plays a role in cellular attachment and entry by HHV-7; this hypothesis will be tested experimentally. First, we will examine the interaction of purified recombinant HHV-7 gp65 with cell surface HSPGs found on cultured human SG cells. Second, a gp65- deleted recombinant virus will be constructed, and its ability to attach to and enter cultured SG cells will be examined. Third, the molecular architecture of gp65, and its interaction with host macromolecules will be studied. In the third aim, we will study two putative 7-transmembrane (7- tm) receptors encoded by HHV-7, U12 and U51. Homologous genes encoded by rat and mouse cytomegalovirus (CMV) are essential for efficient viral replication in salivary glands, and a related gene in human CMV has been shown to contribute to membrane fusion events that may be involved in virus entry or spread. Experiments will therefore be conducted, to determine whether HHV-7 U12 and U51 can enhance membrane fusion events mediated by different viral proteins in cultured SG cells. It is expected that a greater understanding of the molecular pathways exploited by HHV-7 will contribute to the future design of enhanced gene delivery vehicles for SG gene therapy; such vector systems may incorporate components of HHV-7.

人类疱疹病毒（HHV）-7感染与病毒基因组在唾液腺（SG）组织中的持续存在以及病毒抗原在这些部位的慢性表达有关。 该病毒通常被认为是通过唾液途径传播的，并且唾液中存在大量感染性病毒粒子的终身脱落。 这些观察结果表明以下假设：HHV-7已经进化出特定的机制，以进入SG细胞并逃避SG组织中的宿主免疫反应。在这个提议中，我们将研究特定的病毒蛋白在病毒附着和进入SG上皮细胞中的作用。在前两个具体目标中，将研究独特的病毒糖蛋白gp 65。 Gp 65是病毒颗粒的组分，针对gp 65的多克隆抗血清中和病毒感染性; HHV-7 gp 65还结合硫酸乙酰肝素蛋白聚糖（HSPG）。 这些数据有力地表明，gp 65在细胞附着和HHV-7进入细胞中起作用;这一假设将通过实验进行验证。 首先，我们将研究纯化的重组HHV-7 gp 65与在培养的人SG细胞上发现的细胞表面HSPG的相互作用。 其次，将构建gp 65缺失的重组病毒，并检查其附着和进入培养的SG细胞的能力。 第三，研究gp 65的分子结构及其与宿主大分子的相互作用。在第三个目标，我们将研究两个假定的7-跨膜（7- TM）受体编码的HHV-7，U12和U 51。 大鼠和小鼠巨细胞病毒（CMV）编码的同源基因对于唾液腺中的病毒有效复制至关重要，而人类巨细胞病毒中的一个相关基因已被证明有助于膜融合事件，该事件可能参与病毒进入或传播。因此，将进行实验，以确定HHV-7 U12和U 51是否可以增强培养的SG细胞中由不同病毒蛋白介导的膜融合事件。预计对HHV-7所利用的分子途径的更深入了解将有助于未来设计用于SG基因治疗的增强型基因递送载体;此类载体系统可能包含HHV-7的组分。