STUDIES OF THE AMASTIGOTE STAGE OF TRYPANOSOMA CRUZI

几乎完全没有关于使用 克氏锥虫无鞭毛体阶段作为抗原物质的来源 用于抗寄生虫的免疫和用于免疫诊断测定。 我们建议进行研究，以提供这些资料。 到 检查抗性的诱导，无鞭毛体的制备， 化学方法，通过超声处理裂解，或通过使用紫外线灭活 光，将用于免疫小鼠。 不同阶段的准备工作 中医cruzi，epimastigote，同样获得的无鞭毛体 将用于比较结果。 无鞭毛体阶段抗原 以及它们对于免疫诊断测定和 免疫检查。 无鞭毛体抗原的分离将是 通过亲和色谱法用不同的凝集素进行， 特异性和单克隆抗体。 许多单克隆 仅与无鞭毛体阶段存在的抗原反应的抗体 在我们的实验室里。 分离的抗原的能力， 用于诊断恰加斯病的免疫测定将通过 使用免疫诊断技术。 将特别强调 酶免疫测定和抗原吸附的凝集试验 乳胶颗粒 这些分析可以通过使用 纯化的抗原制剂。 免疫反应的类型由 从无鞭毛体中分离的抗原将通过技术 被设计来测量细胞介导的和抗体的反应。 的能力 的分离的抗原，以诱导对T. 将通过用纯化的 单独的抗原和与生物体或化合物的组合， 佐剂，并且已知可增强免疫应答。 拟议 研究将提供有关无鞭毛体阶段的重要且所需的数据 中医cruzi，它积极参与的发病机制， 寄生虫感染，但没有得到足够的重视， 科学家与T. cruzi和恰加斯病。

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.