Tick Vaccine Prevention of Lyme Borreliosis

蜱疫苗预防莱姆疏螺旋体病

• 批准号: 6606563
• 负责人: FRED S KANTOR
• 金额: $ 44.84万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-15 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6606563
• 关键词: Borrelia; Drosophilidae; Escherichia coli; Ixodes; Lyme disease; active immunization; active sites; anticoagulants; bacterial antigens; bacterial vaccines; biotechnology; communicable disease transmission; cooperative study; genetic library; guinea pigs; laboratory mouse; molecular cloning; proteomics; recombinant proteins; saliva; salivary glands; vaccine development

DESCRIPTION (provided by applicant): Our strategy is to prevent transmission of Lyme borreliosis by interfering with the tick's ability to feed to repletion. The proposed project attempts to identify and characterize a family of anticoagulants used by Ixodes ticks needed to permit feeding on their mammalian hosts. Such anticoagulants are vital to the success of tick feeding and in the transmission on tick borne pathogens. We shall explore the family of Ixodes tick anticoagulants by biochemical separation from tick salivary gland and saliva, and by probing cDNA and genomic libraries with appropriate primer fragments. Previous attempts in our laboratories to isolate tick salivary proteins by probing cDNA libraries with antibodies from animals made tick immune resulted in 40 clones; 22 of which were represented by an anticoagulant that we have called SALP-14. In the present proposal we will probe libraries with primers derived from homologous portions of the clones previously identified in order to characterize what we now recognize as a family of anticoagulants vital to tick feeding and transmission of Lyme borreliosis. We will express and purify the recombinant anticoagulants and characterize their activities. To determine active sites, we will generate deletion mutants of these anticoagulants and identify the smallest region essential for effective catalytic activity, as well as antigenicity. The goal is to interfere with feeding by the immune response directed against the functional epitope(s) of the anticoagulant molecules. Feeding to repletion is essential for ticks to complete their life cycle and for the transmission of the pathogens they harbor.

描述(由申请人提供)：我们的策略是通过干扰扁虱的喂养能力来防止莱姆疏螺旋体病的传播。这项拟议的项目试图确定和描述硬蜱所使用的一系列抗凝血剂，以允许它们以哺乳动物宿主为食。这类抗凝剂对成功喂养硬虱和传播硬虱传播病原体至关重要。我们将通过从硬蜱的唾液腺和唾液中进行生化分离，并用合适的引物片段探测cDNA库和基因组文库，来探索硬蜱抗凝剂家族。我们实验室以前的尝试是通过用动物的抗体探测cdna文库来分离壁虱唾液蛋白，结果得到了40个克隆；其中22个克隆由一种我们称为Salp-14的抗凝剂代表。在本提案中，我们将使用从先前鉴定的克隆的同源部分获得的引物来探测文库，以便鉴定我们现在认识到的一类抗凝剂，这些抗凝剂对蜱类饲养和莱姆疏螺旋体病的传播至关重要。我们将对重组抗凝剂进行表达和纯化，并对其活性进行鉴定。为了确定活性部位，我们将产生这些抗凝血剂的缺失突变体，并确定有效催化活性和抗原性所必需的最小区域。其目的是通过针对抗凝剂分子的功能表位(S)的免疫反应来干扰喂养。摄食以补充食物对于硬蜱完成其生命周期和它们所携带的病原体的传播是至关重要的。