LEISHMANIA ANTIGENS--BIOCHEMICAL AND IMMUNOLOGIC STUDIES

这项提议的目的是继续并扩大调查范围 在上一期支持中启动，重点是GP46/M-2基因 利什马尼亚家族。目标应确定免疫学 GP46/M-2诱导保护性免疫的机制 膜表面糖蛋白。基于这样一个事实，即使用 GP46/M-2似乎影响早期感染阶段(S)(导致 无论是发病延迟还是完全保护)，这些研究将集中于 日本血吸虫病早期免疫应答事件的研究 免疫/未免疫小鼠。此外，这项提案试图利用 最新的遗传和生化方法来确定 GP46/M-2家族在体内的生物学作用具体地说 建议的试验方法如下： 1.确定负责的效应细胞、介体和机制 预防因接种GP46/M-2疫苗而引起的感染 佐剂，短小棒状杆菌。试验性方法将采用 细胞重建(使用克隆的T细胞系)和 感染部位的免疫组织化学分析。表位映射 将确定在多肽亚基中潜在使用的保护性表位 疫苗。 2.确定涉及的机制(S)(细胞亚群、细胞因子) 由于使用GP46/M-2-重组痘苗病毒免疫而产生保护作用 病毒。重组GP46-痘苗病毒诱导表达能力的研究 对梅杰乳杆菌和多诺瓦尼乳杆菌感染的保护也将是 已评估。此外，重组非致病基因的潜力 牛痘和金丝雀痘病毒诱导保护性免疫 将调查对利什曼原虫感染的反应。 3.研究结构(生化)、亚细胞分布、 基因调控(在信使核糖核酸水平)和不同物种的生物学功能 GP46/M-2基因家族的成员。功能研究，雇用一名 分子遗传学方法，将重点放在寄生虫的发育上 在血吸虫沙蝇媒介内。

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.