MECHANISMS AND EVALUATION OF VACCINE NEUROVIRULENCE IN VIVO AND IN VITRO

疫苗体内外神经毒力的机制和评价

• 批准号: 6161229
• 负责人: K. M CARBONE
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6161229
• 关键词: brain disorders; developmental neurobiology; disease /disorder model; genetic strain; microorganism growth; mumps virus; neurotropic virus; tissue /cell culture; viral vaccines; virulence; virus replication

Identification of intracellular and cell surface molecules responsible for susceptibility to neurovirulent wild type and vaccine viruses will provide information to help genetically engineer non-neurovirulent vaccines. Using a human cell line that differentiates from fibroblast to neuron (NT2 ) we will examine neuron-specific components linked to neurovirulence. Before and after differentiation, NT2 cells will be infected with parental or neurovirulent variants of non-neurovirulent (e.g., Jeryl Lynn vaccine), or parental or non-neurovirulent variants of neurovirulent (e.g., Kilham wild type) vaccine and wild type mumps virus. Differences in replication (e.g., virus binding, titer of infectious virions, and viral proteins and RNA) will be compared between parental wild type and vaccine viruses and their variants before (fibroblastic) and after (neuronal) differentiation. Cell surface analysis, cell fractionation and gel shift analysis will identify specific components interacting with neurovirulent viruses after neuronal differentiation of cells. Brain damage due to intrauterine virus infections is the most common congenital defect in children. In addition, the infant's brain continues to develop during the first year of postnatal life. Since the developing nervous system is uniquely sensitive to damage following virus infection, administering neurovirulent vaccines to infants also places the child's CNS at increased risk for vaccine related injury. Mumps virus, and certain strains of mumps vaccine (Urabe Am9, Leningrad 3), are among the most neurotropic of the early childhood viruses; New MMR combinations continue to be proposed that include new strains of mumps vaccine virus. Thus, identification of the specific neuronal components that lead to enhanced susceptibility to neurovirulent viruses can lead to sensitive tests for neurovirulence potential during pre-licensing studies. In addition, the development of molecular, in vitro or small animal models of neurovirulence will lead to cost saving and improved predictability of neurovirulence testing.

负责细胞内和细胞表面分子的鉴定 对神经毒性野生型和疫苗病毒的易感性将提供 帮助基因工程非神经毒性疫苗的信息。 使用从成纤维细胞分化为神经元（NT 2）的人细胞系 我们将研究与神经毒力相关的神经元特异性成分。 在分化之前和之后，NT 2细胞将感染亲本 或非神经毒性的神经毒性变体（例如，Jeryl林恩疫苗）， 或神经毒性的亲本或非神经毒性变体（例如，基勒姆 野生型）疫苗和野生型腮腺炎病毒。 复制的差异 (e.g.,病毒结合、感染性病毒粒子的滴度和病毒蛋白， RNA）将在亲本野生型和疫苗病毒之间进行比较， 它们的变体在（成纤维细胞）分化之前和之后（神经元）分化。 细胞表面分析、细胞分级分离和凝胶位移分析将 鉴定与神经毒性病毒相互作用的特定组分， 细胞的神经元分化。 由于宫内病毒感染引起的脑损伤是最常见的 儿童先天性缺陷。 此外，婴儿的大脑继续 在出生后的第一年发育。 由于显影 神经系统对病毒感染后的损伤特别敏感， 给婴儿接种神经毒性疫苗也会使孩子的 CNS发生疫苗相关损伤的风险增加。 腮腺炎病毒， 流行性腮腺炎疫苗的某些菌株（Urabe Am 9，Leningrad 3）是其中之一， 儿童早期病毒中最具嗜神经性的病毒;新的MMR组合 继续建议包括新的腮腺炎疫苗病毒株。 因此，识别导致神经元损伤的特定神经元成分， 增强对神经毒性病毒的易感性可导致敏感的 在许可前研究期间进行神经毒力潜力测试。在 此外，分子，体外或小动物模型的发展， 神经毒力将导致成本节约和改善的可预测性， 神经毒性测试