CELLULAR RECEPTORS FOR HERPES SIMPLEX VIRUS

单纯疱疹病毒（HSV）是一种主要的人类病原体， 通过pH非依赖性的病毒-细胞级联， 交互. 虽然许多研究都集中在了解 病毒成分，最近的发展允许调查的细胞 有助于HSV进入的成分。 参与的细胞成分 硫酸乙酰肝素（HS）。 组件 负责稳定的附着，无论是HS还是假定的蛋白质 受体，仍有待鉴定。 本提案的目的是 分离、鉴定和表征稳定附着受体 HSV-1在感染进入时使用。 研究设计利用了 最近的鉴定和定性提供了独特的机会 由于进入缺陷而对HSV不敏感的细胞。 其具体目的是：（1）将易感性从高度 易感的人类细胞到低或不易感的细胞来克隆 编码HSV稳定附着受体的基因，（2）以确认 并将来自病毒-细胞印迹的110 kd蛋白表征为生物化学 方法来分离和鉴定蛋白质， 受体，以及（3）使用目的1和 2以产生抗受体抗体来表征受体， 其在HSV进入和向性中作用。 这些不同但互补的， 各种办法应与总体目标协同增效。 他们是 由于缺乏功能性非细胞的细胞的可用性， HSV进入所需的肝素样受体。 确定稳定 HSV-1的附着受体对于a）的长期目标是重要的 了解病毒如何穿过细胞膜启动基因 在自然感染期间表达，B）使用疱疹病毒作为载体， c）设计用于避免HSV和其他病毒感染的策略; 导致发病率和死亡率的主要人类病毒病原体，以及d） 确定跨物种融合和交流的分子机制 生物膜

Herpes simplex virus (HSV) is a major human pathogen that enters susceptible cells through a pH independent cascade of virus-cell interactions. While many studies focus on understanding the function of viral components, recent development allow investigation of the cellular components that contribute to HSV entry. The cellular component involved in initial attachment of virus is heparan sulfate (HS). The component(s) responsible for stable attachment, whether HS or a putative protein receptor, remains to be identified. The objective of this proposal is to isolate, identify and characterize the stable attachment receptor(s) used by HSV-1 during infectious entry. The research design exploits the unique opportunity provided by recent identification and characterization of cells that are poorly susceptible to HSV due to a defect in entry. The specific aims are (1) to transfer susceptibility from highly susceptible human cells to low or not susceptible cells to clone the gene(s) encoding the HSV stable attachment receptor(s), (2) to confirm and characterize a 110 kd protein from virus-cell blots as a biochemical approach to isolate and identify the protein(s) which serve as the receptor on human cells, and (3) to use reagents produced in Aims 1 and 2 to generate anti-receptor antibodies to characterize the receptor and its role in HSV entry and tropism. These different, but complementary, approaches should be synergistic to the overall objective. They are feasible because of availability of cells that lack a functional non- heparin-like receptor required in HSV entry. Identifying a stable attachment receptor for HSV-1 is important to long-term goals of a) understanding how viruses cross the cell membrane to initiate gene expression during natural infection, b) using herpesviruses as vectors, c) designing strategies for circumventing infection of HSV and other major human virus pathogens which cause morbidity and mortality, and d) determining molecular mechanisms for fusion and communication across biological membranes.