ROLE OF TICK SALIVA IN LYME DISEASE AND VACCINE STRATEGY

描述（由申请人提供）：载体-宿主界面处的相互作用 可能是许多节肢动物传播的最关键的 传播感染。我们的研究表明， 黑脚硬蜱（肩突硬蜱）利用唾液操纵宿主 免疫反应的方式，既确保血液喂养的成功，并有利于 莱姆病螺旋体（伯氏疏螺旋体）的存活和传播。 我们已经了解到，这些细菌从蜱唾液中获得线索， 它们的蛋白质表达，可能导致增强的侵袭性或存活 在主机。我们已经成功地发现了几种新的分子， 包括我。肩胛肌唾液抗补体蛋白（Isac）和a因子 Xa抑制性抗凝剂（Ixolaris）和重组蛋白在 生产总的来说，这一进展现在使我们能够开始测试我们的 莱姆病的有效预防策略，以及其他 I.肩胛骨传播的感染，最好是通过操作 宿主对载体唾液组分的免疫应答或唾液诱导的免疫应答 微生物产品 在继续这个项目，我们将测试是否一个协议集成向量 唾液腺基因组学和蛋白质组学可以加速发现和 回收潜在重要的免疫原性分子。大量cDNA 一个I.肩胛肌唾液腺cDNA文库， 全长克隆已经揭示了近1,200个序列和至少476个 基因.我们将开始在重组杆状病毒中表达这些序列 表达系统或将它们克隆到细菌质粒中以产生 候选蛋白质和DNA疫苗。此外，最近的技术进步 在B。burgdorferi基因组学使研究取得了快速进展， 检查B。在存在和不存在的情况下， 没有蜱唾液，或在其他饥饿应激条件下。我们将 通过DNA芯片和二维凝胶电泳测试全基因组分析是否 可以帮助发现潜在的免疫原。候选疫苗将 筛选其中断蜱虫进食或阻断病原体的能力 白足鼠（Peromyscus leucopus）模型中的传播。我们预计 这些研究将引导我们开发出联合收割机 蜱和细菌元素，用于预防莱姆病，并可能更广泛 一系列蜱虫传播的感染

DESCRIPTION (Provided by Applicant): Interactions at the vector-host interface are likely to be the most critical to transmission of many arthropod transmitted infections. Our studies have demonstrated that through the action of their saliva, black-legged ticks (Ixodes scapularis) manipulate the host immune response in a manner that both assures blood feeding success, and favors survival and transmission of Lyme disease spirochetes (Borrelia burgdorferi). We have learned that these bacteria receive cues from tick saliva to regulate their protein expression, perhaps leading to enhanced invasiveness or survival in the host. We have been successful in discovering several novel molecules, including I. scapularis' salivary anti-complement protein (Isac) and a Factor Xa-inhibiting anticoagulant (Ixolaris), and recombinant proteins are in production. Taken together, this progress now allows us to begin testing our hypothesis, that an effective prevention strategy for Lyme disease, and other I. scapularis-transmitted infections, can best be developed by manipulating host immune responses to components of vector saliva or saliva-induced microbial products. In continuing this project, we will test whether a protocol integrating vector salivary gland genomics and proteomics can accelerate both discovery and recovery of potentially important immunogenic molecules. Massive cDNA sequencing of an I. scapularis salivary gland cDNA library containing full-length clones has already revealed nearly 1,200 sequences and at least 476 genes. We will begin expressing these sequences in recombinant baculovirus expression systems or cloning them into bacterial plasmids to generate candidate protein and DNA vaccines. In addition, recent technological advances in B. burgdorferi genomics are allowing rapid progress to be made on studies examining B. burgdorferi gene and protein expression in the presence and absence of tick saliva, or under other starvation-stress conditions. We will test whether whole genome analysis by DNA arrays and 2-D gel electrophoresis can facilitate discovery of potential immunogens. Candidate vaccines will be screened for their ability to interrupt tick feeding or block pathogen transmission in a white-footed mouse (Peromyscus leucopus) model. We expect that these studies will lead us to develop vaccination strategies that combine tick and bacterial elements for preventing Lyme disease, and possibly a broader range of tick-transmitted infections.