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

行动 7-免疫反应

• 批准号: 8166220
• 负责人: IDALI MARTINEZ-MARTINEZ
• 金额: $ 10.18万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8166220
• 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; pathogen; 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.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 西尼罗河病毒（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，这种病毒引起了12，000多例报告的人类病例，临床症状从轻度发热疾病到可能致命的脑炎不等。关于西尼罗河病毒在登革热流行地区，如中美洲和南美洲以及加勒比岛屿的传播情况知之甚少。动物流行病研究表明，该病毒在马、鸟类和可能的人类中传播，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性，我们的假设是对登革病毒（DENV）免疫的个体可能对WNV疾病具有交叉保护作用。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用DENV或DEN-2 DNA疫苗候选物主动免疫（具体目标2）或用从原发性或继发性DENV感染收集的人血清被动免疫（具体目标3）来实现针对WNV的交叉保护。针对DENV和WNV的免疫应答将使用Bio-plex系统通过间接免疫荧光测定、中和试验和细胞因子测定来测量。WNV攻击后的小鼠发病率和死亡率将监测28天，血液和脑中的病毒载量将通过Taqman测定法测定。如果实现了交叉保护，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DEN病毒蛋白质，可能在交叉保护中发挥重要作用。这项研究和未来猴子研究的结果将为改进广谱疫苗的设计提供有价值的信息，这些疫苗可能保护免受病原体DENV和WNV的侵害。