COCCIDIOIDES IMMITIS ANTIGENS AS VACCINES

该计划的总体目标是定义免疫球虫抗原，这些抗原是 在实验性球孢子菌病中的有效疫苗，以及可能的 球孢子菌病。由于T细胞介导的免疫在 球孢子菌病，我们建议测试那些能引起强烈T- 细胞反应。这个项目的作用本质上是双重的： 在小鼠身上进行免疫学测试，选择候选抗原 免疫保护实验；并测试这些抗原作为疫苗在 老鼠。我们将使用双管齐下的方法来寻找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 刺激弓形虫特异的小鼠T细胞系，以及C. 两种热休克蛋白HSP60和DNAJ的亚基同源物。这些遗嘱 被检测是否有能力刺激T细胞在淋巴结内的增殖 来自免疫小鼠的细胞和几个球体特异性T细胞系。这些 还将测试它们为小鼠免疫一种增殖剂的能力 对免疫球囊线虫的反应。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 它们会产生淋巴因子。第二种补充方法将是测试 球体来源的T细胞反应性的cDNA库。克隆人 表达的免疫球虫蛋白将在“裸载体”中表达，并且 被DNA“感染”的小鼠。外周血淋巴细胞的增殖反应 小鼠将被检测，那些能引起T细胞反应的将被检测 以重组蛋白的形式表达，并进行如上所述的检测。这个 T细胞反应的大小，引起良好反应的能力 以及产生的淋巴因子将是 决定对哪些抗原进行免疫保护测试。 将在两个遗传菌株中进行免疫保护测试 易受感染的小鼠。将首先测试单个蛋白质，然后再测试 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。

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.