LEISHMANIA ANTIGENS--BIOCHEMICAL AND IMMUNOLOGIC STUDIES

利什曼原虫抗原——生物化学和免疫学研究

• 批准号: 2062052
• 负责人: Diane McMahon Pratt
• 金额: $ 31.75万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-06-01 至 1998-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2062052
• 关键词: Leishmania; active immunization; cell population study; clone cells; cytokine; epitope mapping; genetic manipulation; genetic regulation; glycoproteins; immunocytochemistry; immunomodulators; intracellular parasitism; laboratory mouse; laboratory rabbit; leishmaniasis; life cycle; membrane proteins; microorganism genetics; microorganism immunology; molecular genetics; protozoal antigen; protozoal vaccine; vaccinia virus

The purpose of this proposal is to continue and extend the investigations initiated in the previous period of support, focusing on the GP46/M-2 gene family of Leishmania. The objectives should determine the immunological mechanisms involved in protective immunity elicited by the GP46/M-2 membrane surface glycoprotein. Based on the fact that immunization with GP46/M-2 appears to effect an early phase(s) of infection (resulting in either a delay in onset or complete protection), these studies will focus on the early immunological events occurring in response to infection in immunized/non-immunized mice. Further, this proposal seeks to employ recent genetic as well as biochemical approaches to determine the biological role of the GP46/M-2 family in vivo. Specifically the experimental approaches proposed are: 1. To determine the effector cells, mediators and mechanisms responsible for protection against infection induced by immunization with GP46/M-2 and the adjuvant, Corynebacterium parvum. Experimental approaches will employ cellular reconstitution (using cloned T cell lines) and immunohistochemical analyses at the site of infection. Epitope mapping will determine protective epitopes for potential use in a peptide subunit vaccine. 2. To determine the mechanism(s) (cell subpopulations, cytokines) involved in protection due to immunization using GP46/M-2-vaccinia recombinant viruses. The ability of recombinant GP46-vaccinia virus to induce protection against infection with L. major and L. donovani will also be evaluated. In addition, the potential of recombinant non-pathogenic vaccinia and canary pox viruses in the induction of a protective immune response against infection with Leishmania species will be investigated. 3. To investigate the structure (biochemical), subcellular distribution, gene regulation (at the mRNA level) and biological function of various members of the GP46/M-2 gene family. Functional studies, employing a molecular genetic approach, will focus on development of the parasite within the phlebotomine sand fly vector.

这项建议的目的是继续和扩大调查 在上一个支持期启动，重点是GP 46/M-2基因 利什曼原虫家族目标应确定免疫学 GP 46/M-2诱导的保护性免疫机制 膜表面糖蛋白基于免疫接种 GP 46/M-2似乎影响感染的早期阶段（导致感染）。 无论是延迟发作或完全保护），这些研究将重点 对感染后发生的早期免疫反应， 免疫/非免疫小鼠。此外，该提案旨在利用 最近的遗传学和生物化学方法来确定 GP 46/M-2家族在体内的生物学作用。特别是 提出的实验方法是： 1.确定效应细胞、介质和机制 用于保护免受由GP 46/M-2免疫诱导的感染，和 佐剂，短小棒状杆菌。实验方法将采用 细胞重建（使用克隆的T细胞系）和 免疫组织化学分析在感染部位。表位作图 将确定在肽亚基中潜在使用的保护性表位 疫苗 2.确定涉及的机制（细胞亚群、细胞因子） 在由于使用GP 46/M-2-牛痘重组体免疫而产生的保护中， 病毒重组GP 46-牛痘病毒诱导人胚肺成纤维细胞增殖的能力 预防感染L. major和L.多诺万尼也将是 评估。此外，重组非致病性 牛痘和金丝雀痘病毒诱导保护性免疫 将研究抗利什曼原虫属感染的应答。 3.研究其结构（生化）、亚细胞分布， 基因调控（在mRNA水平）和生物学功能的各种 GP 46/M-2基因家族的成员。功能研究，采用 分子遗传学方法，将集中在寄生虫的发展， 在白蛉媒介中。