ICAM-1 RECEPTOR ANALOGUES AS ANTIRHINOVIRUS AGENTS

鼻病毒的主要组是负责36至45%的所有 普通感冒，数以千万计的学校和工作日，数十亿美元， 去看医生和非处方药的花费， 继发性细菌感染和呼吸道感染加重 疾病 我们将使用细胞受体的主要群体， 鼻病毒，ICAM-1，开发用于病毒治疗的新药， 预防。 ICAM-1是一种单链分子， 免疫球蛋白样结构域和膜锚。 我们先前已经 显示了重组5结构域可溶性ICAM-1抑制病毒结合 和细胞病变效应，并且两个N-末端Ig样 结构域是病毒结合位点正确构象所必需的， 第一域。 测试不同的真核生物和原核生物 表达系统，我们将确定最佳截断位置， 获得重组双结构域ICAM-1片段的条件， 抗病毒活性，并将检测非糖基化和糖基化 衍生物. 最适合结晶的材料将是 大量生产用于测定其三维 结构 这些信息将有助于小分子药物的设计 模拟ICAM-1并占据其在病毒上的结合位点。 多价， 将构建更高亲合力的ICAM-1免疫粘附素嵌合体， 将ICAM-1两个或五个Ig样结构域剪接到IgM、IgA 1 或IgG 1。 假设将被测试，多价赋予高 与病毒的亲和力结合以及中和和诱导蚀变的能力。 低浓度。 电子显微镜将被用来观察 ICAM-1铰链区的位置，免疫粘附素的形状， 与病毒的相互作用。 病毒进入宿主细胞的机制将 研究，假设测试，多价受体结合 和酸性pH代表了内吞过程中触发病毒的事件 渗透和脱膜。 Eclipse是一种不可逆的机制， 病毒中和，假设将被测试，这是 ICAM-1的失活机制，多价ICAM-1是 上级。 鼻病毒的可溶性受体抗性突变体将是 孤立的，假设测试，这些发生在低得多的 频率，如果有的话，多价重组ICAM-1， 紧密模拟细胞表面上展示的受体的亲合力。 讨论了进一步研究和与NIAID工作人员合作的计划。

The major group of rhinoviruses are responsible for 36 to 45% of all common colds, tens of millions of lost school and work days, billions of dollars of doctor's office visits and over-the-counter remedies, secondary bacterial infections, and exacerbations of respiratory diseases. We will use the cellular receptor for the major group of rhinoviruses, ICAM-1, to develop novel drugs for viral therapy and prophylaxis. ICAM-1 is a single chain molecule with 5 immunoglobulin-like domains and a membrane anchor. We have previously shown that recombinant, 5 domain, soluble ICAM-1 inhibits virus binding and cytopathic effect in vitro, and that the two N-terminal Ig-like domains are required for proper conformation of the virus-binding site in the first domain. Testing different eukaryotic and prokaryotic expression systems, we will determine the optimal truncation position and conditions for obtaining a recombinant two domain ICAM-1 fragment with anti-viral activity, and will test both nonglycosylated and glycosylated derivatives. Material that is optimal for crystallization will be produced in large quantity for determination of its three dimensional structure. This information will be useful for design of small drugs that mimic ICAM-1 and occupy its binding site on the virus. Multivalent, higher avidity ICAM-1 immunoadhesin chimeras will be constructed that splice two or five Ig-like domains of ICAM-1 to Fc regions of IgM, IgA1, or IgG1. The hypothesis will be tested that multivalenty confers high affinity binding to virus and ability to neutralize and induce eclipse at low concentration. The electron microscope will be used to visualize the location of the hinge region of ICAM-1, the shape of immunoadhesins, and interaction with virus. The mechanism of virus entry of host cells will be studied, and the hypothesis tested that multivalent receptor binding and acidic pH represent the events during endocytosis that trigger virus penetration and uncoating. Eclipse is an irreversible mechanism for virus neutralization, and the hypothesis will be tested that this is the mechanism of inactivation by, ICAM-1, and that multivalent ICAM-1 is superior. Soluble receptor-resistant mutants of rhinovirus will be isolated, and the hypothesis tested that these occur at much lower frequency, if at all, with multivalent recombinant ICAM-1 that more closely mimics the avidity of receptor displayed on the cell surface. Plans for further study and cooperation with NIAID staff are discussed.