Structure based design of recombinant Zika virus antigens for serodiagnosis

用于血清诊断的重组寨卡病毒抗原的基于结构的设计

• 批准号: 9404101
• 负责人: Aravinda M. DeSilva
• 金额: $ 23.33万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-05 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9404101
• 关键词: Acute; Adult; Antibodies; Antibody Response; Antigens; Area; Binding; Biological Assay; Blood; Blood Tests; Blood donor screening; Childhood; Clinical Management; Congenital Abnormality; Contrast Sensitivity; Dangerousness; Dengue; Dengue Virus; Detection; Diagnosis; Diagnostic; Diagnostic tests; Disease; Encephalitis Viruses; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Epidemiology; Epitopes; Exposure to; Family; Female of child bearing age; Flavivirus; Flavivirus Infections; Follow-Up Studies; Future; Glycoproteins; Guillain-Barré Syndrome; Human; Immune; Immune Sera; Immune response; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Laboratories; Lead; Length; Measures; Methods; Microcephaly; Molecular; Molecular Biology; Monitor; Neonatal; Neurologic; North Carolina; Pathogenesis; Pathogenicity; Pattern; Performance; Persons; Population; Positioning Attribute; Prenatal Diagnosis; Protein Engineering; Public Health; Reagent; Recombinant Proteins; Recombinants; Recording of previous events; Risk; Risk Factors; Serological; Serotyping; Serum; Site; Spontaneous abortion; Structure; Surface; Surveillance Program; Symptoms; System; Universities; Vaccinated; Viral; Viral Antigens; Viral Envelope Proteins; Viral Proteins; Virus; Virus Diseases; West Nile virus; Work; Yellow fever virus; Zika Virus; base; cross reactivity; design; env Gene Products; experience; improved; knowledge base; point of care; protein structure; response; screening; transmission process; viral RNA; virus envelope

Abstract This proposal focuses on the rational design of antigens for serodiagnosis of Zika virus (ZIKV) with minimal cross-reactivity to other human pathogenic flaviviruses. Zika virus (ZIKV) infection can cause miscarriage, microcephaly, and Guillain-Barré syndrome. A specific serodiagnosis of ZIKV infection is urgently needed to track the spread of ZIKV infection and to diagnose individuals exposed to ZIKV in scenarios such as prenatal diagnosis and neonatal monitoring, blood donor screening, screening of travelers returning from Zika-endemic regions. ZIKV serodiagnosis is highly challenging because of extensive cross-reactivity with dengue and other flaviviruses that co-circulate with ZIKV. Our previous and ongoing work defining the human serotype specific antibody responses to dengue envelope (E) protein place us in a unique position to define similar regions in ZIKV E and produce candidate diagnostic antigens for serodiagnostics. Fortified with detailed structural information, we will rationally design and produce substructures of Zika E protein bearing unique antigenic sites for specific and sensitive serological detection of ZIKV infections in flavivirus naïve, flavivirus pre-immune or flavivirus-vaccinated populations. For aim 1, we will produce ZIKV candidate antigens and measure their antibody binding response across a reference panel of immune sera from people exposed to primary or secondary flavivirus infections, including ZIKV and DENV. In Aim 2, we will use protein engineering approaches to redesign most efficacious candidate antigens to change cross-reactive antigenic sites, while preserving Zika specific sites, to improve the diagnostic potential of antigens for differential serodiagnosis of Zika infection. We will compare and contrast one or a combination of surface redesigned Zika antigens for serological detection of recent and prior exposure to ZIKV. In follow up studies, we will partner with reference laboratories and commercial partners to use recombinant protein antigens to develop simple assays for use at the point of care and in surveillance programs.

抽象的 该提案重点介绍了Zika病毒（ZIKV）的抗原的合理设计， 与其他人类病原体的交叉反应性。 Zika病毒（ZIKV）感染会导致流产， 小头畸形和Guillain-Barré综合征。迫切需要对ZIKV感染的特定血清诊断 跟踪ZIKV感染的传播并诊断出在产前等情况下暴露于ZIKV的个体 诊断和新生儿监测，献血者筛查，从Zika-Endimimic返回的旅行者筛查 地区。 ZIKV的血清诊断非常具有挑战性，因为与粉丝和其他 与ZIKV共同循环的黄病毒。我们以前的和正在进行的定义人类血清型的工作 对风扇的抗体反应（E）蛋白将我们置于独特的位置，以定义类似区域 zikv e并产生血清诊断的候选诊断抗原。加强详细的结构 信息，我们将合理设计并生产具有独特抗原的寨卡蛋白的子结构 在黄病毒幼稚的，黄病毒之前的ZIKV感染的特定和敏感的血清学检测部位 或类疫苗接种的人群。对于AIM 1，我们将生产ZIKV候选抗原并测量其 暴露于原发性的人或 继发黄病毒感染，包括Zikv和Denv。在AIM 2中，我们将使用蛋白质工程 重新设计最有效候选者以改变交叉反应性抗原部位的方法，而 保留寨卡病毒特定部位，以提高抗原对差差鉴定血清诊断的诊断潜力 寨卡感染。我们将比较和对比表面重新设计的Zika抗原的一个或组合 近期和事先暴露于ZIKV的血清学检测。在后续研究中，我们将与参考合作 实验室和商业合作伙伴使用重组蛋白抗原来开发简单的测定法 护理点和监视计划。