ACT 7-IMMUNE RESPONSE & OUTCOME OF SEQUENTIAL FLAVIVIRUS INFECTIONS MOUSE MODEL

行动 7-免疫反应

• 批准号: 7959199
• 负责人: IDALI MARTINEZ-MARTINEZ
• 金额: $ 14.65万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959199
• 关键词: Active Immunization; Animals; Antigens; Biological Assay; Birds; Blood; Brain; Clinical; Computer Retrieval of Information on Scientific Projects Database; Country; DNA Vaccines; Dengue; Dengue Virus; Encephalitis; Equus caballus; Fever; Flavivirus Infections; Funding; Future; Grant; Human; Immune; Immune response; Indirect Fluorescent Antibody Technique; Individual; Infection; Institution; Kinetics; Measures; Monitor; Monkeys; Morbidity - disease rate; Mouse Strains; Mus; Neutralization Tests; Outcome; Passive Immunization; Play; Predisposition; Reporting; Research; Research Personnel; Resources; Role; Serum; Source; South America; Symptoms; System; Testing; United States; United States National Institutes of Health; Vaccines; Viral Load result; Viral Proteins; Virus; Virus Diseases; West Indies; West Nile virus; cross reactivity; cytokine; design; epizootic; improved; mortality; mouse model; multidisciplinary; pathogen; programs; research study; transmission process; vaccine candidate

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. West Nile (WNV) is an emerging virus that was introduced in the United States on 1999 and has expanded rapidly throughout the country. This virus caused more than 12,000 reported human cases in the last two years with clinical symptoms that ranged from a mild febrile illness to a potentially fatal encephalitis. Little is known about WNV transmission in dengue-endemic regions, such as Central and South America and the Caribbean islands. Epizootic studies demonstrated that the virus is circulating in horses, birds and probably humans but symptomatic WN cases have not yet been reported in these countries. Since there is extensive cross reactivity between DEN and WNV antigens, our hypothesis is that individuals that are immune to dengue viruses (DENV) may be cross-protected against WNV illnesses. To prove this hypothesis, we will perform animal studies using Balb/c mice. We will first evaluate the susceptibility and kinetics of WNV infection in this mice strain (specific aim 1). Then, two separate experiments will be performed to test if cross-protection against WNV can be achieved by active immunization with DENV or a DEN-2 DNA vaccine candidate (specific aim 2) or passive immunization with human serum collected from primary or secondary DENV infections (specific aim 3). Immune responses against DENV and WNV will be measured by indirect immunofluorescence assays, neutralization tests and cytokine assays using the Bio-plex system. Mice morbidity and mortality after WNV challenge will be monitored for 28 days and viral loads in blood and brain will be determined by Taqman assays. If cross-protection is achieved, these experiments will allow us to determine the type and duration of the immune responses that may correlate with protection. Future studies will be aimed to determine the DEN viral proteins that may play an important role in cross-protection. Results from this and future studies in monkeys will provide valuable information to improve the design of broad spectrum vaccines that could potentially protect against both pathogens, DENV and WNV.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 西尼罗河 (WNV) 是一种新兴病毒，于 1999 年引入美国，并在全国迅速蔓延。在过去两年中，这种病毒导致了超过 12,000 例报告的人类病例，其临床症状包括从轻度发热性疾病到可能致命的脑炎。人们对西尼罗河病毒在登革热流行地区（例如中美洲、南美洲以及加勒比岛屿）的传播知之甚少。动物流行病研究表明，该病毒在马、鸟类和可能人类中传播，但这些国家尚未报告有症状的 WN 病例。由于 DEN 和 WNV 抗原之间存在广泛的交叉反应，我们的假设是，对登革热病毒 (DENV) 免疫的个体可能受到交叉保护，免受 WNV 疾病的侵害。为了证明这一假设，我们将使用 Balb/c 小鼠进行动物研究。我们将首先评估该小鼠品系中 WNV 感染的易感性和动力学（具体目标 1）。然后，将进行两个单独的实验，以测试是否可以通过使用 DENV 或 DEN-2 DNA 候选疫苗进行主动免疫（具体目标 2）或使用从原发性或继发性 DENV 感染收集的人血清进行被动免疫（具体目标 3）来实现针对 WNV 的交叉保护。针对 DENV 和 WNV 的免疫反应将通过使用 Bio-plex 系统的间接免疫荧光测定、中和试验和细胞因子测定来测量。将监测 WNV 攻击后的小鼠发病率和死亡率 28 天，并通过 Taqman 检测确定血液和大脑中的病毒载量。如果实现交叉保护，这些实验将使我们能够确定可能与保护相关的免疫反应的类型和持续时间。未来的研究将旨在确定可能在交叉保护中发挥重要作用的 DEN 病毒蛋白。这项研究和未来的猴子研究结果将为改进广谱疫苗的设计提供有价值的信息，这些疫苗可能会预防登革热病毒和西尼罗河病毒这两种病原体。