Viral receptors of the visual nervous system

描述（由申请人提供）：本提案的目的是扩展我们对呼肠孤病毒3型受体（Reo 3R）的理解：已知与呼肠孤病毒3型血凝素（HA 3）的离散表面结合并参与神经细胞分化和功能的复合物。该受体已被生物化学分离，并发现由两个亚基组成：35-65 kD的糖基化蛋白（CD 147）和95 kD的蛋白质。95 kD亚基是与多跨膜单羧酸转运蛋白受体相关的CD 147复合物。将进行进一步的生物学和生物化学研究以检查CD 147作为呼肠孤病毒3型（T3 D）的体内受体的用途。将进行详细的生化特性和T3 D与该受体结合导致的发病机制的特征。 我们以前研究受体信号传导的努力集中在与这种受体复合物结合的两种结构的表征上。这些结构是HA 3和低亲和力单克隆抗受体抗体MAb 87.92.6，特别是其来自轻链的互补决定区之一。我们已经确定了这些相互作用的许多分子和生物学后果。本课题将重点研究呼肠孤病毒受体的分子分析、通过该受体的信号转导以及其在T3 D新生小鼠神经感染发病机制中的作用。我们将采用一套新的试剂，包括多克隆和单克隆抗体，用于这种分析。这些试剂以高亲和力结合Reo 3R并阻断T3 D附着和随后的感染。 此外，我们将继续努力确定Reo 3R在神经细胞发育中的生物学作用。我们将继续使用高亲和力单克隆抗体来确定受体在发育中的视神经系统中的功能及其在视网膜的Muller和神经节细胞以及在松果体中的特殊作用。我们将研究两种类型的转基因动物：CD 147-/-小鼠和通过使用感光细胞间结合蛋白（IRBP）启动子与识别Reo 3R的MAb轻链（87.92.6L）偶联产生的转基因动物。在表达IRBP-87.92.6L的啮齿动物中，松果体和一些视网膜细胞被遗传消融。我们将在这些突变小鼠中研究T3 D神经发病机制，并定义Reo 3R复合物在影响松果体和视网膜神经发育中的作用原理。

DESCRIPTION (provided by applicant): The objective of this proposal is to extend our understanding of the reovirus type 3 receptor (Reo3R): a complex known to bind to a discrete surface of the reovirus type 3 hemagglutinin (HA3) and to be involved in neural cell differentiation and function. The receptor has been biochemically isolated and found to be composed of two subunits: a 35-65 kD glycosylated protein (CD147) and a 95 kD protein. The 95 kD subunit is a complex of CD147 associated with a multiple membrane-spanning monocarboxylate transporter receptor. Further biological and biochemical studies will be undertaken to examine the use of CD147 as a receptor for reovirus type 3 (T3D) in vivo. Detailed characterization of the biochemistry and features of pathogenesis resulting from binding of T3D to this receptor will be undertaken. Our previous efforts to study receptor signaling focused on the characterization of two structures that bind to this receptor complex. These structures are HA3 and the low affinity monoclonal anti-receptor antibody MAb 87.92.6, particularly one of its complementarity-determining regions from the light chain. We have defined many of the molecular and biological consequences of these interactions. In this proposal, we will focus on the molecular analysis of the reovirus receptor, signal transduction through the receptor, as well as its role in the pathogenesis of neural infection of newborn mice with T3D. We will employ a new set of reagents, including polyclonal and monoclonal antibodies, for this analysis. These reagents bind with high affinity to the Reo3R and block T3D attachment and subsequent infection. In addition, we will continue our efforts to determine the biological role the Reo3R plays in development of neural cells. We will continue to define the receptor's function in the developing optic nervous system and its special role in MuIler and ganglion cells of the retina and in the pineal gland using the high affinity MAb species. We will study two types of transgenic animals: the CD147-/- mouse and a transgenic created by using the interphotoreceptor binding protein (IRBP) promoter coupled with the MAb light chain that recognizes the Reo3R, 87.92.6L. The pineal gland and some retinal cells are genetically ablated in rodents expressing IRBP-87.92.6L. We will study T3D neural pathogenesis in these mutant mice and also define principles of the role of the Reo3R complex in affecting pineal and retinal nerve development.