STUDIES OF THE AMASTIGOTE STAGE OF TRYPANOSOMA CRUZI

克氏锥虫无鞭毛体阶段的研究

基本信息

批准号：
3128221
负责人：
FAUSTO G ARAUJO
金额：
$ 15.5万
依托单位：
PALO ALTO MEDICAL FOUNDATION RES INST
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-04-01 至 1986-03-31
项目状态：
已结题

项目摘要

There is an almost complete absence of information on the use of the amastigote stage of Trypanosoma cruzi as source of antigenic material for immunization against the parasite and for use in immunodiagnostic assays. We propose to conduct research which will provide such information. To examine the induction of resistance, preparations of amastigotes, killed by chemical means, lysed by sonication, or inactivated by using ultraviolet light, will be used to immunize mice. Preparations of the different stage of T. cruzi, the epimastigote, similarly obtained as for the amastigotes will be used for comparison of results. Antigens of the amastigote stage will be isolated and their usefulness for immunodiagnostic assays and for immunization examined. Isolation of the amastigote antigens will be performed by affinity chromatography with lectins of different specificities and with monoclonal antibodies. A number of monoclonal antibodies that react with antigens present in the amastigote stage only are available in our laboratory. The capacity of the isolated antigens for use in immunoassays for diagnosis of Chagas' disease will be evaluated by employing immunodiagnostic techniques. Particular emphasis will be given to enzyme immunoassays and to the agglutination test with antigen-adsorbed latex particles. These assays can be improved considerably by using purified antigen preparations. The type of the immune response elicited by the antigens isolated from the amastigotes will be determined by techniques devised to measure both cell-mediated and antibody responses. The capacity of the isolated antigens to induce resistance against infection with T. cruzi will be examined by immunization of animals with the purified antigens alone and in combination with organisms or compounds that act as adjuvants and are known to enhance the immune responses. The proposed research will provide significant and needed data on the amastigote stage of T. cruzi which participates actively in the pathogenesis of the infection with the parasite but has not received adequate attention from scientists working with T. cruzi and Chagas' disease.

几乎完全没有关于使用的信息克氏锥虫无鞭毛体阶段作为抗原物质的来源针对寄生虫的免疫并用于免疫诊断测定。我们建议进行研究以提供此类信息。到检查抵抗力的诱导，无鞭毛体的准备，被杀死化学方法，通过超声波裂解，或通过使用紫外线灭活光，将用于免疫小鼠。不同阶段的准备 T. cruzi 的上鞭毛体，与无鞭毛体类似地获得将用于结果比较。无鞭毛体阶段的抗原将被分离并且它们对于免疫诊断测定和用于进行了免疫检查。无鞭毛体抗原的分离将是采用不同凝集素的亲和层析法进行特异性和单克隆抗体。多个单克隆仅与无鞭毛体阶段存在的抗原发生反应的抗体我们的实验室提供。分离的抗原的能力在诊断恰加斯病的免疫测定中的应用将由采用免疫诊断技术。将特别强调酶联免疫分析和抗原吸附凝集试验乳胶颗粒。这些测定可以通过使用显着改进纯化的抗原制剂。引起的免疫反应类型从无鞭毛体中分离的抗原将通过技术确定旨在测量细胞介导的反应和抗体反应。容量分离的抗原诱导对 T 感染的抵抗力。 cruzi 将通过用纯化的动物免疫接种来进行检查单独的抗原以及与充当佐剂并已知可增强免疫反应。拟议的研究将提供有关无鞭毛体阶段的重要且所需的数据 T. cruzi 积极参与该病的发病机制感染了寄生虫，但没有得到足够的重视科学家们致力于研究克氏锥虫和南美锥虫病。