CELLULAR BIOLOGY AND IMMUNOLOGY OF LEISHMANIASIS

我们建议研究利什曼病中宿主和寄生虫的相互作用， 识别寄生虫抗原的长期目标可用于 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 也是理解免疫学和 一般的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 观察)利什曼原虫通过附着单核巨噬细胞 纤维连接蛋白(FN)吸附在其表面。我们将尝试确定FN 用1)表面碘化免疫沉淀法检测寄生虫 抗FN抗体和SDS-PAGE；2)特异性免疫荧光 抗FN抗体；3)免疫电子显微镜检查。我们会寻找证据 通过测量125I-FN与寄生虫的结合和与FN受体的结合 寄生到FN涂层表面。我们将防止寄生虫与 单核细胞和其他具有抗FN抗体的细胞，外源性FN，并通过 移除FN受体。将启动分离FN的研究 前鞭毛体的受体。 将通过检测排出能力来研究细胞免疫。 淋巴结细胞诱导利什曼病/杀伤机制。 托皮卡在体外感染巨噬细胞。要评估的变量包括 抗原特异性、遗传限制和效应器的鉴定 细胞表型。我们将开发单抗来识别抗原 在体外通过阻断效应细胞的功能在该反应中被识别。 随后对这些抗原的分离和纯化可用于 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.