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Trial of recombinant MSP-1 vaccine combined with T lymphocyte triggering factor(TLTF)

重组MSP-1疫苗联合T淋巴细胞触发因子（TLTF）的试验

基本信息

批准号：
12670244
负责人：
FUKUMA Toshihide
金额：
$ 1.28万
依托单位：
Kurume University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670244/
关键词：
malaria vaccine MSP-1 Trypanosoma brucei VSG TLTF

项目摘要

We examined the basic research of malaria vaccicne development using African trypanosome (Trypanosoma brucei). Outlines of the results are as follows.1 : The trypanosome expression vectors were constructed with MSP-1 gene (msp-1). These vectors were transfected into T. brucei procyclic form, and expressions were analysed. However, we could not confirm the expression of MSP-1.It is suggested that msp-1 or MSP-1 may have some responsibility in molecular folding or conformation, for example, because the drug resistance gene included in the vectors were functioned and the other reporter genes constructed into the vectors were expressed as expected.Then, the policy of this project was converted to malaria vaccine development using the adjuvant effect of variable surface glycoproteins (VSG) of the T brucei bloodstream form.2 : The fusion proteins of [Histidine TagIVSG N-terminal conserved domain/MSP-1/VSG C-terminal conserved domain] (VSG-MSP) and [Histidine Tag/MSP-1] were tried to express using vaculovirusinsect cell system. The recombinant fusion proteins were solubilized with guanidine, and were purified using the Ni-agarose affinity column.3 : Mice were immunized by subcutaneous injection of recombinant proteins and were subsequently boosted 3 weeks and 6 weeks after the initial immunization. Three weeks after the final inoculation, all mice were challenged with the parasitized 106 erythrocytes by intraperitoneal injection. As the result, slight preventive effects on 9arasitemia were obserbed when mie were immunized with VSGMSP, though they could not survived the infection.

本文综述了利用非洲布鲁氏锥虫开发疟疾疫苗的基础研究。结果概要如下。1：构建MSP-1基因（MSP-1）的锥虫表达载体。将这些载体转染到布氏弧菌原环型中，分析其表达量。然而，我们无法确认MSP-1的表达。提示msp-1或msp-1可能与分子折叠或构象有关，例如，载体中包含的耐药基因功能正常，载体中构建的其他报告基因表达正常。然后，该项目的政策被转化为利用布鲁氏疟原虫血流形式的可变表面糖蛋白（VSG）的佐剂作用开发疟疾疫苗。2 .利用空泡病毒-昆虫细胞系统，尝试表达[组氨酸TagIVSG n端保守域/MSP-1/VSG c端保守域]（VSG- msp）和[组氨酸Tag/MSP-1]融合蛋白。用胍溶解重组融合蛋白，用ni -琼脂糖亲和柱纯化。3：小鼠皮下注射重组蛋白免疫，并在初次免疫后3周和6周增强免疫。最后一次接种3周后，所有小鼠腹腔注射被寄生的106个红细胞。结果表明，小鼠经VSGMSP免疫后，对寄生虫病有轻微的预防作用，但不能存活。