CELLULAR BIOLOGY AND IMMUNOLOGY OF LEISHMANIASIS

我们建议研究利什曼病的宿主-寄生虫相互作用， 长期目标是鉴定可用于治疗的寄生虫抗原， 疫苗 这些研究的重要性不仅来自公众 利什曼病对某些发展中国家的健康意义，但 也是理解免疫学的极好模型， 细胞内感染的细胞生物学。 我们将测试我们的假设（基于令人信服的初步 利什曼原虫附着于单核吞噬细胞， 纤维连接蛋白（FN）吸附在其表面。 我们将尝试识别FN 通过1）表面碘化，然后免疫沉淀 用抗FN抗体和SDS-PAGE进行免疫荧光染色， 抗FN抗体; 3）免疫电镜。 我们会寻找证据 通过测量125 I-FN与寄生虫的结合以及 寄生虫到FN涂覆的表面。 我们将防止寄生虫与 单核细胞和其他细胞与抗FN抗体，外源性FN，并通过 去除FN受体。 将开始研究，以分离FN 前鞭毛体的受体。 细胞介导的免疫将通过检查引流能力来研究。 淋巴结淋巴细胞诱导L. tropica感染的巨噬细胞。 待评估的变量包括 抗原特异性、遗传限制和效应物鉴定 细胞表型 我们将开发单克隆抗体来识别抗原 通过在体外阻断效应细胞功能在该反应中识别。 这些抗原的后续分离和纯化可用于免疫学中。 疫苗

We propose to study host-parasite interactions in Leishmaniasis, with the long range goal of identifying parasite antigens which could be used in a vaccine. The importance of these studies derives not only from the public health significance of leishmaniasis to certain developing countries, but also as an excellent model for understanding the immunology and cell-biology of intracellular infections generally. We will test our hypothesis (based upon compelling preliminary observations) that Leishmania attach to mononuclear phagocytes via fibronectin (FN) absorbed on their surface. We will attempt to identify FN on the parasites by 1) surface iodination followed by immunoprecipitation with anti-FN antibody and SDS-PAGE; 2) immunofluorescence with specific anti-FN antibody; and 3) immune electron microscopy. We will seek evidence for a FN receptor by measuring 125I-FN binding to parasites and binding of parasites to FN-coated surfaces. We will prevent binding of parasites to monocytes and other cells with anti-FN antibody, exogenous FN, and by removal of FN receptors. Studies will be initiated to isolate a FN receptor from promastigotes. Cell-mediated immunity will be studied by examining the ability of draining lymph node lymphocytes to induce leishmanistatic/'cidal mechanisms in L. tropica infected macrophages in vitro. Variables to be assessed include antigen specificity, genetic restriction, and identification of effector cell phenotype. We will develop monoclonal antibodies to identify antigens recognized in this reaction by blocking effector cell function in vitro. Subsequent isolation and purification of these antigens could be used in a vaccine.