MOLECULAR ANALYSIS OF POLIOVIRUS NEUTRALIZING EPITOPES

脊髓灰质炎病毒中和表位的分子分析

• 批准号: 3133969
• 负责人: Marie Chow
• 金额: $ 18.34万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3133969
• 关键词: antibody neutralization test; antiserum; antiviral antibody; genetic mapping; immunogenetics; monoclonal antibody; mutagens; mutant; nucleic acid sequence; oligonucleotides; poliomyelitis vaccine; poliovirus; protein sequence; serotyping; transducin; transfection; viral carcinogenesis; virus antigen; virus classification; virus envelope; virus genetics; virus infection mechanism; virus protein; virus replication

The aim of the proposed research is to contribute to a more detailed molecular description of the poliovirus neutralizing sites and the immune response induced by the virus. A combination of genetic and immunological approaches are proposed to test predictions derived from a structural analysis of the neutralizing sites, thereby A) extending the structural description of the previously identified neutralizing sites for poliovirus serotype 1, B) identifying potentially new neutralizing sites, and C) functionally characterizing regions in the capsid proteins that are critical for the viability of the virus. To achieve these goals, several studies are proposed: 1) Viral mutants will be constructed using site-specific mutagenesis methods. The molecular biology and antigenic characteristics of these mutants will be studied. 2) Regions within the capsid proteins that appear to be hypermutable or mutationally constrained will be identified by sequencing six naturally-occuring poliovirus serotype 1 strains. These strains were sequentially isolated over a 13 month period by the Centers for Disease Control from cases that occurred during an epidemic outbreak of paralytic poliomyelitis within the Amish community. This study will indicate the spectrum of amino acid substitutions and the regions within the virus that viable mutations can occur. 3) The immunogenicity of the VP4 capsid protein will be studied. Although the immunogenicity of the other three capsid proteins have been extensively studied, the antibody response to VP4 is uncharacterized and may play important roles in poliovirus pathogenesis. The studies proposed here provide an unique opportunity to extend our understanding beyond the identification of the amino acids that form a neutralizing determinant and to address issues concerning: how antibodies interact with the neutralizing sites, what functional roles do different regions on the virus surface (including the neutralizing sites) serve for the biology of the virus, how viruses mutate to become neutralization resistant and generate antigenic variants, how are new serotypes formed. These issues are important for a thorough understanding of the molecular mechanisms of viral pathogenesis. In addition, the health relatedness of this proposal derives from its contribution to the understanding the molecular basis of host protection by viral vaccines and the potential application of this knowledge to the rational design of vaccines for viruses (e.g. HTLV and HIV strains, and hepatitis A) and for the improvement of existing viral vaccine (such as poliovirus).

拟议研究的目的是促进更多 脊髓灰质炎病毒中和位点的详细分子描述 以及病毒引起的免疫反应。 的组合 建议采用遗传和免疫学方法来测试 中和结构分析得出的预测 站点，从而A）扩展了结构描述 先前确定的脊髓灰质炎病毒血清型 1 的中和位点， B) 识别潜在的新中和位点，以及 C) 衣壳蛋白中的功能特征区域 对于病毒的生存能力至关重要。 为了实现这些目标， 提出了几项研究： 1) 将使用位点特异性构建病毒突变体 诱变方法。 分子生物学和抗原 将研究这些突变体的特征。 2）衣壳蛋白内的区域似乎是 超可变或突变受限将通过以下方式识别 对六种天然存在的脊髓灰质炎病毒血清型 1 菌株进行测序。 这些菌株是在 13 个月的时间内依次分离出来的 疾病控制中心从期间发生的病例 阿米什人中麻痹性脊髓灰质炎的流行 社区。 这项研究将表明氨基酸的谱 替代和病毒内可行的区域 可能会发生突变。 3)研究VP4衣壳蛋白的免疫原性。 虽然其他三种衣壳蛋白的免疫原性 已被广泛研究，对 VP4 的抗体反应是 未表征且可能在脊髓灰质炎病毒中发挥重要作用 发病。 这里提出的研究提供了一个独特的机会来扩展 我们的理解超越了氨基酸的鉴定 形成中和决定因素并解决问题 涉及：抗体如何与中和位点相互作用， 病毒表面不同区域有何功能作用 （包括中和位点）服务于病毒的生物学， 病毒如何突变以产生中和抗性以及 产生抗原变异，新的血清型是如何形成的。 这些问题对于全面了解 病毒发病机制的分子机制。 此外， 该提案的健康相关性源自其对 了解病毒保护宿主的分子基础 疫苗以及这些知识的潜在应用 合理设计病毒疫苗（例如 HTLV 和 HIV 毒株， 和甲型肝炎）以及改进现有病毒疫苗 （如脊髓灰质炎病毒）。