CD1d/NKT-binding Glycolipids as an Adjuvant for a T cell-based Malaria Vaccine

项目摘要 根除造成地方病和大流行病的全球病原体取决于 研制有效的疫苗。疟疾的情况肯定就是这样。然而，我们无法 为了引起强烈和持久的保护性T细胞反应，特别是CD 8 + T细胞反应， 这是成功开发疫苗的主要障碍。因此，佐剂技术将 可能不仅对克服对病毒疫苗载体的预先存在的免疫力至关重要， 进一步增强疫苗免疫原性。我们以前的研究表明，CD 1d 分子结合的自然杀伤T（NKT）细胞配体α-半乳糖神经酰胺（α-GalCer）可以增强 鼠疟疾疫苗引起的保护性CD 8 + T细胞应答，包括重组 表达疟疾抗原的腺病毒。与两个著名的化学团体合作， 在黄志辉博士和理查德·弗兰克博士的指导下，我们成功地鉴定了几种a- 作为NKT细胞配体的GalCer类似物。 在这个提议中，我们的目标是首先筛选一百个α-GalCer类似物的聚焦文库， 我们最近产生的，然后选择一个较小的小组的候选糖脂的基础上， 它们在与鼠或人NKT细胞一起培养时引起的体外细胞因子产生谱。 我们还将确定由所选糖脂引起的体内细胞因子产生概况 给小鼠施用后。这些体外和体内筛选过程将引导我们选择一种 十几种有希望的候选糖脂，显示出强烈的Th 1偏向性、Th 2偏向性或双极性 活动我们的第二个目标是确定每种药物的佐剂效应的大小， 新鉴定的糖脂有助于疟疾疫苗的免疫原性。我们将 随后表征抗疟疾CD 8 + T细胞应答， 一个老鼠模型。我们的第三个也是最后一个目标将是确定器官和细胞类型， 疟疾抗原的CD 8 + T细胞，并随后揭示机制， α-GalCer及其类似物的佐剂作用。这项工作将涉及利用重组 腺病毒，共表达疟疾抗原和绿色荧光蛋白（GFP）。 项目叙述 疟疾继续对全球社会、特别是对成年人构成严重威胁 以及前往或生活在世界热带和亚热带地区的儿童。的 该提案的目的是开发一种新的战略，以提高疟疾的疗效， 通过使用糖脂作为免疫增强化合物或佐剂来制备疫苗。的 开发合适的疟疾疫苗/糖脂佐剂组合将具有 减少疟疾发病率和降低发病率和死亡率的潜力 是由这种病原体引起的

Project Summary The eradication of global pathogens responsible for endemic and pandemic diseases hinges upon the development of effective vaccines. This is certainly the case for malaria. However, our inability to elicit ¿strong and long-lasting¿ protective T cell responses, particularly CD8+ T cell responses, has been a major obstacle to successful vaccine development. Accordingly, adjuvant technologies will likely be critical not only to overcome pre-existing immunity to viral vaccine vectors but also to further enhance vaccine immunogenicity. Our previous studies have demonstrated that a CD1d molecule-binding, natural killer T (NKT) cell ligand, a-galactosylceramide (a-GalCer), can enhance protective CD8+ T cell responses elicited by murine malaria vaccines, including a recombinant adenovirus expressing a malarial antigen. In collaboration with two eminent chemistry groups directed by Dr. Chi-Huey Wong and Dr. Richard Franck, we have successfully identified several a- GalCer analogs that act as NKT cell ligands. In this proposal, we aim to first screen a focused library of one hundred a-GalCer analogs that we have recently generated, and then select a smaller panel of candidate glycolipids based on the in vitro cytokine production profiles they elicit upon cultivation with murine or human NKT cells. We will also determine the in vivo cytokine production profiles elicited by the selected glycolipids upon administration to mice. These in vitro and in vivo screening processes will lead us to choose a dozen of promising candidate glycolipids that display strong Th1-biased, Th2-biased, or bipolar activities. Our second aim will be to determine the magnitude of adjuvant effect that each of these newly identified glycolipids contributes to the immunogenicity of a malaria vaccine. We will subsequently characterize the anti-malarial CD8+ T cell responses augmented by the glycolipids in a mouse model. Our third and final aim will be to determine the organs and cell types that present the malarial antigen to CD8+ T cells, and subsequently to uncover the mechanisms underlying the adjuvant effects of a-GalCer and its analogs. This work will involve the utilization of a recombinant adenovirus, co-expressing a malarial antigen and green fluorescent protein (GFP). Project Narrative Malaria continues to pose a grave threat to the global community and in particular to adults and children traveling to or living in tropical and subtropical regions of the world. The purpose of this proposal is to develop a novel strategy to enhance the efficacy of malaria vaccines by employing glycolipids as immuno-enhancing compounds or ¿adjuvants.¿ The development of a suitable malaria vaccine/glycolipid adjuvant combination would have the potential to decrease the incidence of malaria and diminish the morbidity and mortality attributable to this pathogen.