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.

摘要 该提案的重点是合理设计用于寨卡病毒（ZIKV）血清学诊断的抗原， 与其他人类致病性黄病毒的交叉反应性。寨卡病毒（ZIKV）感染可导致流产， 小头畸形和格林-巴利综合征迫切需要ZIKV感染的特异性血清学诊断， 跟踪ZIKV感染的传播，并诊断在产前、产后和产后等情况下暴露于ZIKV的个体。 诊断和新生儿监测，献血者筛查，从寨卡流行病返回的旅行者筛查 地区ZIKV血清学诊断是高度挑战性的，因为与登革热和其他病原体具有广泛的交叉反应性。 与ZIKV共循环的黄病毒。我们以前和正在进行的工作，确定人类血清型特异性 对登革热包膜（E）蛋白的抗体应答使我们处于独特的位置，以确定 ZIKV E并产生用于血清诊断的候选诊断抗原。详细的结构加固 信息，我们将合理设计和生产具有独特抗原性的Zika E蛋白的亚结构， 在黄病毒初治、黄病毒免疫前和免疫后的ZIKV感染中进行特异性和灵敏性血清学检测的研究中心 或黄病毒疫苗接种人群。对于目标1，我们将产生ZIKV候选抗原并测量它们的表达。 抗体结合反应跨越来自暴露于原发性或继发性免疫缺陷的人的免疫血清的参考组， 继发性黄病毒感染，包括ZIKV和DENV。在目标2中，我们将使用蛋白质工程 重新设计最有效的候选抗原以改变交叉反应性抗原位点的方法， 保留寨卡病毒特异性位点，以提高用于寨卡病毒鉴别血清学诊断的抗原的诊断潜力， 寨卡病毒感染我们将比较和对比表面重新设计的寨卡抗原之一或组合， 最近和先前暴露于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开发简单的检测方法， 护理点和监控项目。