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性反应的可能性导致了这样的概念，即有效的疫苗应该针对无鞭毛体阶段特异性抗原。本研究的目的是表征在无鞭毛体阶段特异性表达的蛋白质，并在无鞭毛体阶段特异性蛋白质中找到潜在的抗原作为疫苗候选物。1）证实了无鞭毛体在皮肤利什曼病发病中的重要作用，即感染部位宿主细胞（主要是巨噬细胞）的浸润和无鞭毛体在巨噬细胞内的增殖是致病的主要原因。2)建立了一种简便的实验小鼠无鞭毛体纯化方法。3)证实了无鞭毛体和前鞭毛体在BALB/cA小鼠中的感染性和蛋白质表达水平方面的差异。4)2D SDS-PAGE分析表明过氧化物酶是一种蛋白质，在无鞭毛体中表达。过氧化物酶可能作为抗氧化应激的最后一道防线发挥重要作用。此外，还制备了与无鞭毛体有较强反应性的无鞭毛体阶段特异性单克隆抗体，用于无鞭毛体的详细研究。这些研究将为利什曼病的免疫诊断、治疗和有效疫苗的研制提供新的思路，即以无鞭毛体阶段特异性抗原为控制措施。

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。

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 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）

Okuno T.et al.: "Applications of recombinant Leishmania amazonensis expressing egfp or the b-galactosidase gene of for drug screening and histopathological analysis."Experimental Animals. 52. 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。