Tick Vaccine Prevention of Lyme Borreliosis

蜱疫苗预防莱姆疏螺旋体病

• 批准号: 7021410
• 负责人: FRED S KANTOR
• 金额: $ 47.85万
• 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 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7021410
• 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tick的喂食能力来防止莱姆毛毛虫病传播。拟议的项目试图识别和表征由ixodes tick虫使用的一系列抗凝剂家族，以便允许哺乳动物寄主。这样的抗凝剂对于滴答作用的成功和tick病原体的传播至关重要。我们将通过与滴答唾液腺和唾液的生化分离以及探测具有适当的底漆片段的cDNA和基因组库来探索ixodes家族抗凝剂的家族。以前的实验室尝试通过用动物的抗体探测cDNA库来分离tick唾液蛋白，从而使tick免疫产生了40个克隆。其中22个由我们称为salp-14的抗凝剂代表。在本提案中，我们将探测以前鉴定出的克隆同源部分的底漆，以表征我们现在认识到的抗凝剂家族，这对于滴答了滴答lyme borreliosis的滴答至关重要。我们将表达和净化重组抗凝剂并表征其活动。为了确定活性位点，我们将产生这些抗凝剂的缺失突变体，并确定有效催化活性以及抗原性所必需的最小区域。目的是干扰针对抗凝分子的功能表位的免疫反应的喂养。为了完成其生命周期和传播其携带的病原体的滴答至关重要。