Defining Molecular Epitopes of Protective Antibodies to Flaviviruses

项目摘要/摘要： 本次K08临床科学家导师职业奖申请的目标是为Alena Markmann博士提供 具有雄厚的人体免疫学基础和以前研究人类免疫细胞的最新方法 开始了她在人类对RNA病毒的免疫反应方面的独立研究生涯。马克曼博士是 北卡罗来纳大学教堂山分校医学助理教授，终身教职 医学博士，微生物学和免疫学系的联合任命。候选人有 在抗体结构、抗体-抗原相互作用和 病毒的抗体中和。通过加强她对RNA病毒的适应性免疫和 掌握了分析抗原特异性B细胞的最新方法，马克曼博士将研究人类 对登革热和寨卡病毒等新出现的黄病毒的免疫反应可以抑制病毒感染和疾病或 加强病毒复制，加剧疾病。这位候选人和她的导师阿拉文达·德席尔瓦博士 设计了一项培训计划，其中包括严格的研究部分以及教学指导、网络 和演示机会、团队管理技能以及手稿写作和资质。 这些技能将共同确立成功的职业发展所必需的方法和原则。 黄病毒是一种包膜的正链RNA病毒，对人类种群构成日益增长的威胁。 每年造成数百万人感染。它们是由媒介传播的，由扁虱和蚊子传播，并引起 一系列疾病表现，包括发烧疾病，可导致终生的脑炎 并发症、先天性寨卡综合征、登革热出血热和休克综合征。最多的 部分，黄病毒感染导致对感染病毒的中和保护性抗体反应，这可以 感染后可持续数年。然而，这四种登革热病毒在抗原性上非常相似，在许多情况下， 导致对个体具有的登革热病毒血清型产生广泛的交叉中和抗体反应 还没有接触到。到目前为止，还没有人确定这些广泛中和登革热的病毒靶点。 存在于血清中的抗体。此外，尽管我们知道寨卡病毒的几个主要保护目标 从研究记忆B细胞的抗体反应来看，我们不知道血清的靶标是什么。初步结果 提示在寨卡病毒血清抗体反应的情况下，结合靶点与已知的强相关 中和记忆B细胞来源的抗体靶点。因此，我们在目标1中假设，血清抗体 反应将反映记忆B细胞的反应，但可能来自较少数量的B细胞克隆和 减少与其他黄病毒的交叉反应。在目标2中，我们将确定广泛交叉的病毒结合靶点 中和血清中的登革热抗体。这些研究将导致对关键保护的认同 针对登革热病毒和寨卡病毒的血清抗体靶点，以告知成功 疫苗设计。

Project Summary/Abstract: The objective of this K08 Mentored Clinical Scientist Career Award Application is to provide Dr. Alena Markmann with a strong foundation in human immunology and the latest methods for studying human immune cells before launching her independent research career on the human immune response to RNA viruses. Dr. Markmann is an Assistant Professor of Medicine, tenure track, at the University of North Carolina -Chapel Hill (UNC) School of Medicine with a joint appointment in the Department of Microbiology and Immunology. The candidate has expertise in and a strong track record of publications on antibody structure, antibody-antigen interactions and antibody neutralization of viruses. By strengthening her knowledge of adaptive immunity to RNA viruses and mastering the latest methods for analyzing antigen specific B cells, Dr. Markmann will study how the human immune response to emerging flaviviruses like dengue and Zika can suppress viral infection and disease or enhance viral replication and exacerbate disease. The candidate and her mentor Dr. Aravinda de Silva have designed a training plan that includes a rigorous research component along with didactic instruction, networking and presentation opportunities, team management skills as well as manuscript writing and grantsmanship. Together these skills will establish the methods and principles necessary for successful career development. Flaviviruses are enveloped positive stranded RNA viruses that pose a growing threat to the human population and cause millions of infections annually. They are vector-borne, transmitted by ticks and mosquitoes, and cause a spectrum of disease manifestations including febrile illness, encephalitides that can cause lifelong complications, Congenital Zika Syndrome and dengue hemorrhagic fever and shock syndromes. For the most part, flaviviral infection result in a neutralizing protective antibody response against the infecting virus, that can last for years after infection. The four dengue viruses however are very similar antigenically and in many cases, result in generating a broad cross-neutralizing antibody responseto dengue virus serotypes that individuals have not yet been exposed to. Thus far, no one has identified the viral target site of these broadly neutralizing dengue antibodies that exist in the serum. Furthermore, though we know of a few major targets of the Zika protective antibody response from studying memory B cells, we do not know what the serum targets are. Preliminary results suggest that in the case of Zika serum antibody responses, binding targets are correlated with known strongly neutralizing memory B cell-derived antibody targets. Thus, we hypothesize in Aim 1, that serum antibody responses will mirror memory B cell responses, but will likely come from a smaller number of B cell clones and be less cross-reactive with other flaviviruses. In Aim 2 we will identify the viral binding targets of broadly cross- neutralizing dengue antibodies from the serum. These studies will result in the ident ification of critical protective viral targets to both dengue and Zika viruses from the serum antibody compartment in order to inform successful vaccine design.