COCCIDIOIDES IMMITIS ANTIGENS AS VACCINES

犬球孢子菌抗原作为疫苗

• 批准号: 6493572
• 负责人: Theo N Kirkland
• 金额: $ 15.71万
• 依托单位: VETERANS MEDICAL RESEARCH FDN/SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6493572
• 关键词: Coccidioides immitis; T lymphocyte; cellular immunity; clinical research; coccidioidomycosis; disease /disorder model; fungal antigens; fungal proteins; fungal vaccines; human tissue; laboratory mouse; leukocyte activation /transformation; lymphokines; recombinant proteins; tissue /cell culture; vaccine development; vector vaccine

The overall goal of this program is to define C. immitis antigens which are effective vaccines in experimental coccidioidomycosis, and presumably, hman coccidioidomycosis. Since T- cell mediated immunity is critical in coccidioidomycosis, we propose to test antigens which elicit vigorous T- cell responses. The role of this project is essentially twofold: to perform the immunologic testing in mice, selecting candidate antigens for immunoprotection experiments; and to test those antigens as vaccines in mice. We will use a two-pronged approach to find T-cell reactive C. immitis antigens. We have already identified and cloned one protein which stimulated a C. immitis-specific murine T-cell line, as well as the C. immitis homologs of two heat-shock proteins, hsp 60 and dnaJ. These will be tested for ability to stimulate T-cell proliferation in lymph node T- cells from immune mice and several spherule-specific T-cell lines. These will also be tested for their ability to immunize mice for a proliferative response to C. immitis spherules. In collaboration with Project 3, we will determine what type of T-cells are activated by these antigens and what lymphokines they produce. A second, complementary approach will be to test cDNA libraries derived from spherules for T-cell reactivity. The clones expressing C. immitis proteins will be expressed in "naked vectors" and the mice "infected" with the DNA. The T-cell proliferative responses of the mice will be determined, and those that elicit T-cell responses will be expressed as recombinant proteins and tested as outlined above. The magnitude of the T-cell responses, the ability to elicit good responses against intact spherules, and the lymphokines produced will be factors in deciding which antigens will be tested for immunoprotection. Immunoprotection assays will be done in two strains of genetically susceptible mice. Single proteins will be tested first and then combinations. We anticipate that these studies will provide information critical to the development of a human vaccine.

这个程序的总体目标是定义C。免疫性抗原， 有效的疫苗在实验球孢子菌病，并推测，人类 球孢子菌病 由于T细胞介导的免疫对于 球孢子菌病，我们建议测试抗原，引起强烈的T- 细胞反应。 该项目的作用基本上是双重的： 在小鼠中进行免疫测试，选择候选抗原， 免疫保护实验;并测试这些抗原作为疫苗， 小鼠 我们将使用双管齐下的方法来发现T细胞反应性C。 免疫性抗原 我们已经鉴定并克隆了一种蛋白质， 刺激了C. immitis特异性鼠T细胞系，以及C. 两种热休克蛋白hsp 60和dnaJ的immitis同源物。 这些将 测试刺激淋巴结T细胞增殖的能力， 来自免疫小鼠的细胞和几种球特异性T细胞系。 这些 还将测试它们免疫小鼠的增殖能力， 回应C。肠炎小球。 与项目3合作，我们将 确定哪些类型的T细胞被这些抗原激活， 它们产生的淋巴因子。 第二种补充方法是测试 用于T细胞反应性的源自小球的cDNA文库。 克隆人 表达C.免疫蛋白将在“裸载体”中表达， 被DNA“感染”的老鼠 T细胞增殖反应的研究 小鼠将被确定，那些引发T细胞反应的小鼠将被 表达为重组蛋白并如上所述进行测试。 的 T细胞反应的大小，引发良好反应的能力 对抗完整的小球，产生的淋巴因子将是 决定哪些抗原将被测试用于免疫保护。 免疫保护试验将在两种遗传性 易感小鼠 首先测试单个蛋白质，然后 组合。 我们预计这些研究将提供信息， 对人类疫苗的发展至关重要。