ROLE OF CELL INTEGRINS IN ADENOVIRAL CONJUNCTIVITIS

腺病毒（Ad）是引起结膜炎的主要原因， 伙计 虽然通常是自限性的，但Ad眼部感染具有很高的 传染性水平和传播率， 由于大量工作日损失而导致的发病率和社会经济问题 每年.在眼部感染期间发生的免疫反应也可以 导致严重视觉障碍，包括失明。 旨在阻断病毒进入细胞的抗病毒策略已经被广泛应用。 由于缺乏精确的知识， 介导进入事件的病毒和细胞成分。细胞 表面整合素α-v-β 3和α-v-β 5介导腺病毒 通过特异性相互作用内化而不是病毒附着 在病毒五邻体基础衣壳蛋白中具有RGD序列。此外，委员会认为， 我们最近的研究表明，整合素α-M-β 2和α-L- β 2促进腺病毒附着于人单核细胞和淋巴细胞 细胞本建议旨在利用这些研究结果， 体内阻断腺病毒眼部感染的合理方法。 由于几种不同的腺病毒血清型使用α-V整联蛋白用于 感染时，使用这些受体的强效拮抗剂可赋予 针对多种病毒血清型的保护。 一系列详细的分子和生物化学研究将用于 阐明Ad五邻体碱基与不同细胞的精确相互作用 整合素，以获得对腺病毒细胞进入的进一步了解。 五邻体碱结合的动力学、热力学和化学计量学 将使用自动化生物传感器测定细胞整合素 系统五邻体碱基中精确的氨基酸序列 介导的与不同细胞整联蛋白的结合将通过 使用激光分析肽与固定化整联蛋白的结合， 解吸质谱法该提案的第二个目标是 确定β 2整合素在腺病毒感染中的总体作用 人单核细胞/巨噬细胞和淋巴细胞，两种细胞类型， 有助于Ad眼部发病机制。这些细胞也可能是 在宿主中持续性Ad感染的位点。最后，一个强大的 细胞整联蛋白或功能阻断剂的合成肽拮抗剂 单克隆抗体，其识别多个 腺病毒血清型将用于预防腺病毒感染， NZW兔眼模型。 这些研究为阻断Ad的眼部作用提供了一条合理的途径 体内感染，也可能导致改善的策略， 用于眼部基因治疗的复制缺陷型腺病毒载体。

Adenoviruses (Ad) represent a major cause of viral conjunctivitis in man. Although usually self-limiting, Ad ocular infections have a high level of infectivity and transmission rate causing significant morbidity and socioeconomic problems due to numerous lost working days each year. Immune responses occurring during ocular infections can also lead to severe visual disturbances including blindness. Antiviral strategies aimed at blocking virus entry into cells have been relatively underdeveloped due to a lack of knowledge of the precise viral and cellular components that mediate the entry events. The cell surface integrins alpha-v-beta3 and alpha-v-beta5 mediate adenovirus internalization rather than virus attachment via a specific interaction with an RGD sequence in the virus penton base capsid protein. Moreover, our recent studies indicate that integrins alpha-M-beta2 and alpha-L- beta2 promote adenovirus attachment to human monocytic and lymphoid cells. This proposal seeks to capitalize on these findings to develop a rational approach to blocking adenovirus ocular infections in vivo. Since several different adenovirus serotypes use alpha-v integrins for infection, the use of potent antagonists of these receptors may confer protection against multiple virus serotypes. A series of detailed molecular and biochemical studies will be used to elucidate the precise interactions of Ad penton base with distinct cell integrins in order to gain further insights into adenovirus cell entry. The kinetics, thermodynamics and stoichiometry of penton base binding to cell integrins will be determined using an automated biosensor system. The precise amino acid sequences in the penton base that mediate binding to different cell integrins will be identified by analyzing peptide binding to immobilized integrins using laser- desorption mass spectrometry. A second goal of the proposal is to determine the overall role of beta2 integrins in adenovirus infection of human monocyte/macrophages and lymphocytes, two cell types that contribute to Ad ocular pathogenesis. These cells are also a likely site for persistent Ad infection in the host. Finally, a potent synthetic peptide antagonist of cell integrins or a function-blocking monoclonal antibody that recognizes the RGD domain of multiple adenovirus serotypes will be used to prevent adenovirus infection in the NZW rabbit ocular model. These studies represent a rational approach to block Ad ocular infection in vivo and may also lead to improved strategies for using replication-defective adenovirus vectors for ocular gene therapy.