Peptide-vaccine development against Lyme disease

项目摘要 莱姆病是一种地方性蜱传疾病，与关节炎等衰弱表现相关， 肌肉疼痛、心脏炎、脑膜炎和脑脊髓炎。尽管在这一领域作出了广泛努力， 预防这种感染的疫苗可供人类使用。为了解决这个问题，我们的实验室 开发了新的方法来鉴定蜱和哺乳动物细菌感染期间相关的抗原， 基于这些蛋白质的胞外和保守区域设计肽抗原。 我们对这项工作的假设是，基于外膜蛋白的肽在细胞的各个阶段表达， B的地方性循环。伯氏疏螺旋体可用作针对莱姆病的保护性疫苗抗原。测试 因此，我们建议对B进行RNA测序。在感染蜱和小鼠的过程中， 在两种生物体中表达最高的基因（Aim 1）。我们的目标是合成肽基于 这些蛋白质的细胞外区域，并确定其在鼠模型中的抗原性。我们会选择抗原 其是高度保守的、免疫原性的，并且不显示与人抗原的交叉反应性，以避免脱- 目标反应性（目标2）。然后，我们将在小鼠疫苗接种模型中测试这些抗原， 与能够引发特异性体液和细胞免疫应答的各种佐剂组合。做 为此，我们将使用IFN-γ、IL-6和IL-17的各种鼠报告系统来测量细菌清除率， 免疫和攻击期间的T细胞应答（目的3）。为了选择最好的佐剂，我们还将测量 生发中心的形成和记忆B和T细胞的产生。在这些研究结束时，我们将有 配制含有基于外膜蛋白的抗原和佐剂的基于肽的疫苗 能够产生强烈的细胞免疫反应。这些研究将有助于对小说的鉴别 抗原，并为预防莱姆病提供答案。

Project Summary Lyme disease is an endemic tick-borne disease associated with debilitating manifestations such as arthritis, muscle pain, carditis, meningitis, and encephalomyelitis. Despite extensive efforts in the field, there is still no vaccine for the prevention of this infection available for human use. To address this problem, our laboratory has developed novel methodologies to identify antigens relevant during bacterial infection in ticks and mammals and design peptide antigens based on extracellular and conserved regions of these proteins. Our hypothesis for this work is that peptides based on outer-membrane proteins expressed in various phases of the enzootic cycle of B. burgdorferi can be used as protective vaccines antigens against Lyme disease. To test this, we propose to perform RNA sequencing of B. burgdorferi during infection of ticks and mice to identify the genes most highly expressed in both organisms (Aim 1). Our objective is to synthesize peptides based on the extracellular regions of these proteins, and determine their antigenicity in a murine model. We will select antigens that are highly conserved, immunogenic, and do not display cross-reactivity with human antigens to avoid off- target reactivity (Aim 2). We will then test these antigens in a murine model of vaccination and challenge in combination with various adjuvants capable of eliciting specific humoral and cellular immune responses. To do this, we will use various murine reporter systems for IFN-γ, IL-6 and IL-17 to measure bacterial clearance and T-cell response during immunization and challenge (Aim 3). To select the best adjuvant, we will also measure germinal center formation and memory B and T cell production. At the conclusion of these studies, we will have formulated a peptide-based vaccine containing antigens based on outer-membrane proteins with an adjuvant capable of conferring a strong cellular immune response. These studies will help with the identification of novel antigens and provide answers for the prevention of Lyme disease.