Neurological Disorder: Role of the UOL in CNS Immune System

神经系统疾病：UOL 在 CNS 免疫系统中的作用

• 批准号: 7047973
• 负责人: GUEY-CHUEN PERNG
• 金额: $ 30.98万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7047973
• 关键词: Adhesives; Adult; Affect; Afferent Neurons; Animals; Apoptosis; Axon; Biological Assay; Blindness; Brain; Cell Communication; Cells; Cerebrum; Cessation of life; Chimeric Proteins; Clinical; Co-Immunoprecipitations; Complex; Condition; Confocal Microscopy; Data; Development; Doxycycline; Encephalitis; Eye; Genes; Genomics; Goals; Herpesvirus 1; Immune; Immune response; Immune system; In Vitro; Incidence; Infection; Intercellular Adhesion Molecules; Lead; Lesion; Leukocytes; Light; Location; Lymphocyte; Molecular; Monoclonal Antibodies; Mus; Neurons; Oligonucleotides; Pathogenesis; Play; Protein Binding; Protein Overexpression; Proteins; Publications; Recurrence; Reporting; Research Personnel; Ribonucleases; Role; Serological; Simplexvirus; Staining method; Stains; Stress; Structure of trigeminal ganglion; Survival Rate; Symptoms; System; Tamoxifen; Testing; Thick; Time; Transcript; Transgenes; Transgenic Organisms; Travel; Viral; Virulence; Virus; Virus Replication; Western Blotting; base; cDNA Library; cell type; corneal scar; cryostat; cytokine; in vivo; mutant; nervous system disorder; neurotropic; neurovirulence; programs; research study; response; telencephalin; yeast two hybrid system

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) infects the eyes, replicates and travels through axons to the sensory neurons, where it becomes latent. Sporadically the virus can reactivate, return to the eye and cause corneal scarring and loss of vision. We have cloned a new HSV-1 gene, designated UOL, that is located in a genomic region previously suggested to be involved in viral virulence. Based upon two-hybrid system preliminary studies, UOL protein interacts strongly with ICAM-5 (intercellular adhesion molecules 5) from mouse brain. ICAM-5 is an important molecule in neuron-leukocyte interactions that is proposed to attract immune cells to the CMS. In pull-down experiments using lysates from mouse brains, UOL is able to pull-down ICAM-5. In addition, delayed immune response in immune lymphocytes infiltrating to CNS in wild type compared to that of UOL deletion virus infected mice. Thus, we hypothesize that UOL-ICAM-5 interactions play an important role in HSV virulence. To test this hypothesis, we propose the following specific aims: (1) Confirm that UOL-protein binds ICAM-5 in vitro and in vivo by protein-protein pull-down assays, co-immunoprecipitation, and confocal microscopy. (2)Confirm that UOL-ICAM-5 interactions are involved in HSV-1 neurovirulence. Several viral mutants will be constructed including anti-sense ICAM-5 oligonucleotides that can block ICAM-5 expression, UOL-shut off, and UOL-turn on mutants. By blocking ICAM-5 will significantly increase virulence of the HSV UOL mutant, while over express UOL should produce increased virulence. (3) Dissect the effects of immune cells by the interaction of UOL with ICAM-5 in CNS. ICAM-5 regulates immune response in CNS on the setting of infection. Immune lymphocytes infiltrating into CNS will be characterized and cytokines profile will be documented. Taken together, theses aims will shed new light on how HSV evades CNS immune system and may lead to the development of new treatments for HSV encephalitis.

描述（由申请人提供）：单纯疱疹病毒（HSV）感染眼睛，复制并通过轴突传播到感觉神经元，在那里它变得潜伏。偶尔，病毒会重新激活，回到眼睛，导致角膜瘢痕和视力丧失。我们克隆了一个新的HSV-1基因，命名为UOL，该基因位于先前认为与病毒毒力有关的基因组区域。基于双杂交系统的初步研究，UOL蛋白与小鼠脑内细胞间粘附分子ICAM-5 （intercell adhesion molecules 5）有较强的相互作用。ICAM-5是神经元-白细胞相互作用中的一个重要分子，被认为可以吸引免疫细胞到CMS。在使用小鼠大脑裂解物的下拉实验中，UOL能够下拉ICAM-5。此外，与UOL缺失病毒感染小鼠相比，野生型小鼠浸润到中枢神经系统的免疫淋巴细胞的免疫反应延迟。因此，我们假设UOL-ICAM-5相互作用在HSV毒力中起重要作用。为了验证这一假设，我们提出了以下具体目标：(1)通过蛋白-蛋白拉下实验、共免疫沉淀和共聚焦显微镜证实uol蛋白在体外和体内与ICAM-5结合。(2)证实UOL-ICAM-5相互作用参与HSV-1的神经毒力。将构建几种病毒突变体，包括可以阻断ICAM-5表达的反义ICAM-5寡核苷酸，uol关闭和uol打开突变体。通过阻断ICAM-5可显著提高HSV UOL突变体的毒力，而过表达UOL则可提高其毒力。(3)通过UOL与ICAM-5在中枢神经系统的相互作用，解剖免疫细胞的作用。ICAM-5在感染时调节中枢神经系统的免疫反应。免疫淋巴细胞浸润中枢神经系统将被表征，细胞因子谱将被记录。综上所述，这些目标将揭示HSV如何逃避中枢神经系统免疫系统，并可能导致HSV脑炎新疗法的发展。