Enhanced immunogenicity and protection study of lipid modified protein vaccine candidates of Nontypeable Haemophilus influenzae

摘要： 在结合疫苗时代，非分型流感嗜血杆菌(NTHi)已成为 年急性中耳炎、复发性中耳炎、急性鼻窦炎和结膜炎的主要病因 年儿童和成人与慢性阻塞性肺疾病(COPD)急性加重 成年人。有必要开发一种针对NTHi的疫苗。各种候选疫苗已经被 探索但仍需加强NTHi抗体和Th17免疫应答 将有助于防止鼻咽定植和感染的疫苗。在这个项目中，我们将 重组蛋白P6和OMP26及其融合构建体免疫原性的比较 在NTHi-AOM小鼠混合感染模型中，我们建立了它们的脂化和非脂化形式。 根据它们不同的功能选择了两种蛋白质，并考虑到P6是自然的 脂化的天然OMP26不是但已知能诱导Th17免疫。我们将测试我们的 假设脂化蛋白抗原也会引起更高的血液和粘膜抗体水平 作为Th17免疫反应(通过Toll样受体2激活)，而不是非脂蛋白。理想化 与蛋白质抗原相比，蛋白质抗原将更大程度地减少耳朵和鼻腔的细菌负荷 抗NTHi的非脂化抗原。抗体介导的免疫球蛋白的作用及机制 在幼年和成年小鼠中，将比较Th17介导的免疫对NTHi的保护作用 在使用脂化、非脂化和融合构建的疫苗的TLR2基因敲除小鼠中也是如此 考生P6和OMP26。我们将测试脂化蛋白是否会产生更强大的IL-17A 鼻咽部记忆Th17细胞的反应，帮助运输中性粒细胞和显示 与非脂化蛋白抗原相比，NTHi清除增强。整体而言，建议的 研究将极大地促进我们对重组人免疫原性增强的理解 TLR-2依赖的蛋白质及其脂肪调节的分子机制 Th17-NTHi疫苗研制中的免疫。

Abstract: In the conjugate vaccine era, Nontypeable Haemophilus influenzae (NTHi) has become the leading cause of acute otitis media (AOM), recurrent AOM, acute sinusitis and conjunctivitis in children and adults and acute exacerbations of chronic obstructive pulmonary disease (COPD) in adults. There is need to develop a vaccine against NTHi. Various vaccine candidates have been explored but still there is need for enhancement of antibody and Th17 immunity response for NTHi vaccines that will help prevent nasopharyngeal colonization and infection. In this project we will compare immunogenicity of recombinant proteins P6 and OMP26 and their fusion constructs in their lipidated and non-lipidated form in a mouse coinfection model of NTHi-AOM, we developed. Two proteins were selected based on their different function and considering P6 is naturally lipidated and native OMP26 is not but is known to induce Th17 immunity. We will test our hypothesis that lipidated protein antigens elicit higher blood and mucosal antibody levels as well as Th17 immune response (via toll-like receptor 2 activation) than non lipidated proteins. Lipidated protein antigens will elicit greater reduction in ear and nasal bacterial loads compared to nonlipidated antigens against NTHi. The contribution and mechanism of antibody mediated and TH17-mediated immunity in protection against NTHi will be compared in infant and adult mice as well as in TLR2-knock out mice using lipidated, nonlipidated and fusion constructs of vaccine candidates P6 and OMP26. We will test whether lipidated proteins produce a more robust IL-17A response from memory Th17 cells in the nasopharynx that helps traffic neutrophils and show enhanced NTHi clearance compared to nonlipidated protein antigens. Overall, the proposed studies will significantly advance our understanding of enhanced immunogenicity of recombinant proteins and molecular mechanisms underlying the lipidation regulation of TLR-2 dependent Th17- immunity in NTHi vaccine development.