Functional screening for neutralizing antibodies targeting CMV entry factors

描述（由申请方提供）：人巨细胞病毒（CMV）是一种在人群中血清阳性率为60-90%的疱疹病毒，可导致免疫功能低下个体的发病率和死亡率。发生在免疫功能低下宿主中的CMV疾病的常见表现包括婴儿中的神经元缺陷和移植受者中的胃肠道疾病、肺炎、CMV综合征和终末器官疾病。值得注意的是，巨细胞病毒是影响全球2.5%新生儿的出生缺陷的主要原因，并且与早期血管损伤有关，因为它可以感染内皮细胞和巨噬细胞。CMV感染多种细胞类型如内皮细胞、上皮细胞和单核细胞的能力对于宿主内的病毒传播和增殖是必不可少的。CMV进入需要病毒与细胞表面结合，结合后受体聚集，随后病毒与细胞膜融合并在胞质溶胶中释放衣壳。病毒融合事件可以发生在细胞表面以及以pH依赖性方式发生在上皮细胞和内皮细胞的内体内。尽管表皮生长因子受体和血小板源性生长因子受体与病毒进入有关，但CMV进入的复杂性将需要许多额外的细胞因子来确保成功的病毒感染。我们假设，独特的细胞表面蛋白是成功的病毒感染所必需的病毒结合，内吞作用，融合，运输到细胞核，和病毒基因表达中发挥重要作用。因此，我们计划利用最近开发的中和抗体的功能筛选，以发现对CMV进入重要的上皮细胞和内皮细胞因子。抗体靶点的鉴定和表征将揭示新的进入因子，并提供对病毒感染早期阶段的分子理解。此外，这些试剂将允许同时开发抗CMV治疗剂，其可用于限制CMV相关病症高风险的免疫受损患者中的CMV疾病。

DESCRIPTION (provided by applicant): Human cytomegalovirus (CMV) is a ¿-herpesvirus with a seroprevalance of 60-90% among the population that can cause morbidity and mortality in immuno-compromised individuals. Common manifestations of CMV disease that occur in immunocompromised hosts include neuronal defects in infants and gastrointestinal disorders, pneumonia, CMV syndrome, and end-organ disease in transplant recipients. Remarkably, CMV is the leading cause of birth defects affecting up to 2.5% of newborns worldwide and is linked to early vascular damage because it can infect endothelial cells and macrophages. The ability of CMV to infect multiple cell types such as endothelial cells, epithelial cells, and monocytes is essential for viral dissemination and proliferation within the host. CMV entry requires viral binding to the cell surface, post-binding receptor clustering followed by viral fusion with cell membranes and release of capsid in the cytosol. The viral fusion event can occur at the cell surface as well as in a pH-dependent manner within endosomes of epithelial and endothelial cells. Even though the epidermal growth factor receptor and the platelet-derived growth factor receptor-¿ have been implicated in virus entry, the given complexity of CMV entry would require many additional cell factors to ensure a successful virus infection. We hypothesize that unique cell surface proteins are essential for a successful virus infection by playing important roles in virus binding, endocytosis, fusion, trafficking to nucleus, and viral gene expression. Thus, we plan to utilize a recently developed functional screen for neutralizing antibodies to discover epithelial and endothelial cell factors important for CMV entry. The identification and characterization of the antibody's target will uncover novel entry factors and provide a molecular understanding of the early phase of a virus infection. Also, these reagents will allow for the simultaneous development of anti-CMV therapeutics that can be used to limit CMV disease in immuno- compromised patient at high-risk for CMV-associated disorders.