ICAM-1 RECEPTOR ANALOGUES AS ANTI-RHINOVIRUS AGENTS

鼻病毒的主要群体占所有病毒总数的36%到45% 普通感冒，数以千万计的学校和工作日的损失，数十亿 去看医生的钱和非处方药， 继发性细菌感染和呼吸道疾病加重 疾病。我们将对主要人群使用细胞受体 鼻病毒，ICAM-1，开发病毒治疗的新药和 预防措施。ICAM-1是一种单链分子，含有5 免疫球蛋白样结构域和膜锚定。我们之前已经 显示重组的5个结构域的可溶性ICAM-1抑制病毒结合 和体外的细胞病变效应，以及两个N端的Ig样 结构域是正确构象病毒结合部位所必需的。 第一个领域。检测不同的真核生物和原核生物 表达系统中，我们将确定最优截断位置和 重组双结构域ICAM-1片段的获得条件 抗病毒活性，并将测试非糖基化和糖基化 衍生品。最适合结晶的材料将是 为确定其三维尺寸而大量生产 结构。这些信息将对小型药物的设计有用 它模仿ICAM-1并占据其在病毒上的结合位置。多价， 将构建亲和力更高的ICAM-1免疫粘附素嵌合体 将ICAM-1的两个或五个Ig样结构域与IgM，IgA1， 或者是IgG1。这一假设将得到检验，即多价给予高 与病毒的亲和力以及中和和诱导日食的能力 低浓度。电子显微镜将被用来显示 ICAM-1铰链区的位置，免疫粘附素的形状，以及 与病毒的相互作用。病毒进入宿主细胞的机制将 进行了研究，该假说检验了多价受体结合 和酸性pH代表内吞过程中触发病毒的事件 渗透和脱涂层。日食是一种不可逆转的机制 病毒中和，这一假设将得到检验，这是 ICAM-1的失活机制，以及多价ICAM-1是 苏必利尔。鼻病毒的可溶性受体耐药突变株将是 孤立的，并且假设检验了这些发生在低得多的地方 如果有的话，多价重组ICAM-1的频率更高 与显示在细胞表面的受体亲和力非常接近。 讨论了与NIAID工作人员进一步学习和合作的计划。

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.