Viral receptors of the visual nervous system

描述(申请人提供)：这项建议的目的是扩大我们对呼肠孤病毒3型受体(Reo3R)的理解：一种已知与3型呼肠孤病毒血凝素(HA3)的离散表面结合并参与神经细胞分化和功能的复合体。该受体已被生化分离，并发现由两个亚基组成：35-65kD糖基化蛋白(CD147)和95kD蛋白。95 kD亚基是CD147的复合体，与多个跨膜的单羧酸转运蛋白受体相关。将进行进一步的生物学和生化研究，以检查CD147在体内作为3型呼肠孤病毒(T3D)受体的用途。将对T3D与该受体结合所导致的生化和致病特征进行详细的表征。 我们以前研究受体信号的努力集中在与这个受体复合体结合的两个结构的表征上。这些结构是HA3和低亲和力的抗受体单抗MAb 87.92.6，特别是它的轻链互补决定区域之一。我们已经定义了这些相互作用的许多分子和生物学后果。在这项建议中，我们将集中在呼肠孤病毒受体的分子分析，通过该受体的信号转导，以及它在T3D新生小鼠神经感染发病机制中的作用。我们将使用一套新的试剂，包括多克隆和单克隆抗体，用于这一分析。这些试剂与Reo3R具有高亲和力，并阻止T3D附着和随后的感染。 此外，我们将继续努力确定Reo3R在神经细胞发育中所起的生物学作用。我们将继续使用高亲和力的单抗来确定受体在发育中的视神经系统中的功能以及它在视网膜的Muiller细胞和神经节细胞以及松果体中的特殊作用。我们将研究两种类型的转基因动物：CD147-/-小鼠和使用光受体间结合蛋白(IRBP)启动子与识别Reo3R的单抗轻链耦合创建的转基因动物，87.92.6L。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基因消融。我们将研究这些突变小鼠的T3D神经发病机制，并确定Reo3R复合体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.