HSV-Receptor Interaction in Entry and Pathogenesis

描述(申请人提供)：单纯疱疹病毒(HSV)可引起多种人类疾病，包括唇疱疹、眼部和生殖器感染、新生儿感染和脑炎。HSV1和HSV2这两种血清型都会在感觉神经节内建立终生潜伏感染。由于单纯疱疹病毒与宿主有复杂的相互作用，因此包膜包含11种病毒编码的糖蛋白也就不足为奇了。这些蛋白质在病毒入侵、传播、免疫逃避和抑制细胞凋亡方面发挥作用。此外，病毒可以通过几种不同的受体进入细胞。许多HSV1和2毒株可以使用其中的两种，HveA(疱疹病毒进入介质A)和Nectin-1(HveC)。HveA在T淋巴细胞上大量存在，在其他类型的细胞上也有少量存在。Nectin-1是一种细胞黏附分子，在上皮细胞和神经细胞上大量存在。我们的目标是增加我们对Nectin-1和HSV受体结合蛋白gD之间相互作用的了解，并确定这种相互作用在细胞和体内HSV感染中的意义。对Gd单独或与HveA结合的结构的解表明，当Gd与HveA结合时，Gd发生了两个构象变化。我们假设，当GD与Nectin-1结合时，至少有一种情况也会发生，这可能对GD在以后的进入步骤中的作用很重要。因此，解决GD/Nectin-1复合体的结构是该项目的主要目标。Nectin-1可以与自身以及在细胞贴壁连接处与其他Nextin相互作用。我们推测，新合成的GD作为Nectin-1在邻近未感染细胞上的配基，从而确保该受体可用于病毒传播到下一个细胞。我们进一步假设，Nectin-1从受感染的细胞表面下调，因此不会阻止病毒传播到下一个细胞。最后，我们构建了一组具有明确受体用途的gD突变体。我们将把它们重新组合到病毒基因组中，并在小鼠模型中测试它们引起原发带状疱疹病毒病的能力。这项资助的目的是：(1)描述GD和Nextin-1之间的相互作用；(2)探索GD和HSV感染对Nectin-1介导的功能的影响；以及(3)确定Nectin-1和其他HSV进入受体在病毒感染小鼠模型中的作用。

DESCRIPTION (provided by applicant): Herpes simplex viruses (HSVs) cause a variety of human diseases, including cold sores, eye and genital infections, neonatal infections and encephalitis. Both serotypes, HSV1 and HSV2, establish lifelong latent infections within sensory ganglia. Because HSV has a complex interaction with its host, it is not surprising that the envelope contains eleven virus-encoded glycoproteins. These proteins function in virus entry, spread, immune evasion and inhibition of apoptosis. In addition, the virus can enter cells via several different receptors. Many strains of HSV1 and 2 can use two of these, HveA (herpes virus entry mediator A) and nectin-1 (HveC). HveA is found in copious amounts on T lymphocytes and to a lesser extent on other cell types. Nectin-1 is a cell adhesion molecule that is abundant on epithelial and neuronal cells. Our goal is to increase our understanding of the interaction between nectin-1 and gD, the receptor binding protein of HSV and to determine the significance of this interaction for HSV infection in cells and in vivo. Solution of the structure of gD alone or bound to HveA revealed that two conformational changes in gD occur when it binds HveA. We hypothesize that at least one of these also occurs when gD binds nectin-1 and may be important for the role of gD in later steps of entry. Therefore, solving the structure of the gD/nectin-1 complex is a major goal of this project. Nectin-1 can interact in trans with itself as well as with other nectins at cellular adherens junctions. We speculate that newly synthesized gD acts as a ligand for nectin-1 on adjacent uninfected cells, thereby ensuring that receptor is available for the virus to spread to the next cell. We further hypothesize that nectin-1 is down regulated from the infected cell surface and therefore does not impede virus spread to the next cell. Finally, we have constructed a panel of gD mutants with defined receptor usage. We will recombine these into the viral genome and test their ability to cause primary and zosteriform disease in a mouse model. The aims of this grant are: (1) to characterize the interaction between gD and nectin-1; (2) to explore the effects of gD and HSV infection on nectin-1 mediated functions; and (3) to determine the role of nectin-1 and other HSV entry receptors in a mouse model of virus infection.