Development of vaccine and diagnosis targeted on Leishmania amastigotes specific protein

针对利什曼原虫无鞭毛体特异性蛋白的疫苗开发和诊断

• 批准号: 13557020
• 负责人: MATSUMOTO Yoshitsugu
• 金额: $ 3.71万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557020/
• 关键词: Leishmaniasis; Promastigotes; Serodiagnosis; 無鞭毛型原虫; 有鞭毛型原虫

The possibility of a protective response against Leishmania parasites has led to the concept that an effective vaccine should be aimed at amastigote stage-specific antigens. The aim of this study is to characterize proteins that are expressed specifically in the amastigote stage, and to find potential antigens as vaccine candidates among amastigote stage-specific proteins. In this study, 1) The important role of amastigotes was conformed that the pathogenesis in cutaneous leishmaniasis was mainly attributed to the infiltration of host cells, mainly macrophages, at the site of infection and the intramacrophage propagation of amastigotes. 2) A simple purification method for amastigotes from experimental mice was developed. 3) The differences between amastigotes and promastigotes in terms of infectivity in BALB/cA mice and protein expression levels were confirmed. 4) 2D SDS-PAGE analysis revealed that peroxidoxin is a protein, which is expressed in amastigotes. Peroxidoxin might play an important role as the last line of defense against oxidative stress. In addition, amastigote stage-specific monoclonal antibodies, which had strong reactivity with amastigotes, were produced for detailed studies on amastigotes. These studies will contribute to the development of imunological diagnosis or treatment or effective vaccine against leishmaniasis with the new concept that anlastigote stage-specific antigens should be aimed for controlmeasures.

针对利什曼原虫寄生虫的保护反应的可能性导致了这样一个概念，即应采用有效的疫苗针对变性剂阶段特异性抗原。这项研究的目的是表征在amastigote阶段特别表达的蛋白质，并在amastigote阶段特异性蛋白中找到潜在的抗原作为疫苗候选。在这项研究中，1）amastigotes的重要作用是符合皮肤利什曼病的发病机理的主要归因于宿主细胞（主要是巨噬细胞）在感染部位和合子症的摩托结构繁殖位。 2）开发了一种简单的实验小鼠杂物化方法。 3）确认了BALB/CA小鼠的感染性和蛋白质表达水平的杂物和前肢差异。 4）2D SDS-PAGE分析表明，过氧化物毒素是一种蛋白质，在amastigotes中表达。过氧化物毒素可能是针对氧化应激的最后防御措施的重要作用。此外，产生了具有强烈反应性的合并期特异性单克隆抗体，以详细研究杂种。这些研究将有助于进化Imunological诊断或治疗或对利什曼病的有效疫苗，新概念是应针对特异性的抗原抗原。

项目成果

Okuno, T. et aI.: "Applications of recombinant Leishmania amazonensis expressing egfp or the b-galactosidase gene for drug screening and histopathological analysis."Experimental Animals. 52(2). 109-118 (2003)

Okuno, T. 等人：“表达egfp 或b-半乳糖苷酶基因的重组亚马逊利什曼原虫在药物筛选和组织病理学分析中的应用。”实验动物。

Matsumoto Y. et al.: "New epidemics of parasitic diseases attendant on irrigation projects."Journal of Arid Land Studies. 13(3). 201-208 (2003)

Matsumoto Y. 等人：“灌溉项目带来的寄生虫病新流行。”干旱土地研究杂志。

松本 芳嗣 ら: "動物の感染症 Infectious Diseases of Animals"(株)近代出版. 556 (2001)

松本 Yoshitsugu 等：《动物传染病》近代出版有限公司 556（2001）

Hatabu T. et ai.: "The expression system of biologically active canine interleukin-8 in Leishmania promastigotes."Parasitology International. 51. 63-71 (2002)

Hatabu T.等人：“利什曼原虫前鞭毛体中具有生物活性的犬白细胞介素8的表达系统。”国际寄生虫学。

松本 芳嗣: "動物の感染症Infectious Diseases of Animals"(株)近代出版. 556 (2001)

松本义嗣：《动物传染病》近代出版有限公司556（2001）