Peptide-vaccine development against Lyme disease

项目摘要 莱姆病是一种地方性扁虱传播疾病，与关节炎等衰弱症状有关， 肌肉疼痛、心脏炎、脑膜炎和脑脊髓炎。尽管在实地进行了广泛的努力，但仍然没有 预防这种感染的疫苗可供人类使用。为了解决这个问题，我们的实验室已经 开发了新的方法来识别扁虱和哺乳动物中细菌感染过程中相关的抗原，并 根据这些蛋白质的胞外区和保守区设计多肽抗原。 我们对这项工作的假设是，基于外膜蛋白的多肽在不同时期表达 伯氏杆菌的流行循环可作为莱姆病的保护性疫苗抗原。为了测试 为此，我们建议在感染扁虱和小鼠的过程中进行伯氏杆菌的RNA测序，以确定 在两种生物中表达最高的基因(目标1)。我们的目标是合成基于蛋白质的多肽 这些蛋白的胞外区，并在小鼠模型中确定它们的抗原性。我们将选择抗原 高度保守的，免疫原性的，不与人类抗原发生交叉反应的，以避免脱离 目标反应性(目标2)。然后，我们将在疫苗接种的小鼠模型中测试这些抗原，并在 与各种佐剂相结合，能够诱发特定的体液和细胞免疫反应。去做 为此，我们将使用干扰素-γ、IL-6和IL-17的各种小鼠报告系统来测量细菌清除率和 免疫和挑战期间的T细胞反应(目标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、T细胞的产生。在这些研究结束后，我们将拥有 以外膜蛋白为基础，辅以佐剂的多肽疫苗的研制 能够产生强烈的细胞免疫反应的。这些研究将有助于小说的鉴定 抗原，并为莱姆病的预防提供答案。

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.