HSV-Receptor Interaction in Entry and Pathogenesis

描述（由申请人提供）：单纯疱疹病毒（hsv）引起多种人类疾病，包括唇疱疹、眼睛和生殖器感染、新生儿感染和脑炎。两种血清型，HSV1和HSV2，在感觉神经节内建立终身潜伏感染。因为HSV与宿主有复杂的相互作用，所以包膜包含11种病毒编码的糖蛋白并不奇怪。这些蛋白在病毒进入、传播、免疫逃避和抑制细胞凋亡中起作用。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）和连接素-1 （HveC）。HveA在T淋巴细胞中含量丰富，在其他细胞类型中含量较少。Nectin-1是一种细胞粘附分子，在上皮细胞和神经元细胞中含量丰富。我们的目标是增加我们对连接蛋白-1与HSV受体结合蛋白gD之间相互作用的理解，并确定这种相互作用对细胞和体内HSV感染的意义。对gD单独或与HveA结合的结构的分析表明，gD与HveA结合时发生了两种构象变化。我们假设，当gD与连接蛋白-1结合时，这些过程中至少有一个也会发生，并且可能对gD在随后的进入步骤中所起的作用很重要。因此，解决gD/nectin-1复合物的结构是本项目的主要目标。Nectin-1可以反式与自身以及细胞粘附连接处的其他连接蛋白相互作用。我们推测新合成的gD在相邻未感染细胞上作为连接素-1的配体，从而确保受体可用于病毒传播到下一个细胞。我们进一步假设nectin-1在受感染细胞表面被下调，因此不会阻止病毒向下一个细胞扩散。最后，我们构建了一个具有明确受体使用的gD突变体面板。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性和带状虫状疾病的能力。这项资助的目的是：(1)描述gD和nectin-1之间的相互作用；(2)探讨gD和HSV感染对nectin-1介导功能的影响；(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.