Novel malaria transmission-blocking vaccine development using cell-free protein synthesis system

利用无细胞蛋白质合成系统开发新型疟疾传播阻断疫苗

基本信息

批准号：
16017273
负责人：
TSUBOI Takafumi
金额：
$ 9.6万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16017273/
关键词：
malaria transmission-blocking vaccine cell-free protein synthesis system international exchange Thailand

项目摘要

Although not providing direct protection for vaccinees, vaccines targeting antigens expressed on the surface of the sexual stages of the parasites are considered one promising strategy for malaria control, and these malaria transmission-blocking vaccines (TBV) have received increased attention over the past decade. This blocks parasite developmental cycles within the mosquito midgut, consequently reducing or preventing parasite transmission to other individuals. TBVs are also expected to prevent the spread of escape mutants emerged during the course of anti-malaria drug treatment or other prophylactic vaccines targeting asexual stages of the parasites. Only four candidate molecules of this type of vaccine are studied for their potential to induce protective immunity, to date. Since the genomic sequence of P falciparum was completed in October 2002, we have now free access to the genome data to search for novel vaccine candidates. However, one of the bottlenecks for the vaccine development is the high AT content in exons of P. falciparum, which will considerably inhibit the recombinant protein expression using conventional methods. Our strategy is to express recombinant proteins for the characterization of each protein based on the genome sequences of P. falciparum without using synthetic gene. In order to identify the novel TBV candidates, we selected 190 genes, which are expected to be expressed only in gametocyte stage of P falciparum. These genes were cloned into plasmids and templates were prepared for transcription through PCR-based procedures, followed by high throughput recombinant protein synthesis by wheat germ cell-free system. Using this approach, we succeeded in obtaining 120 recombinant proteins. After the screening of these recombinant proteins to identify novel TBV candidates with transmission-blocking human sera, we have identified 15 novel gametocyte antigens as TBV candidates.

尽管没有为疫苗提供直接保护，但靶向在寄生虫性阶段表面表达的抗原的疫苗被认为是疟疾控制的一种有前途的策略，而这些疟疾传播阻断疫苗（TBV）在过去十年中受到了越来越多的关注。这阻断了蚊子中肠内的寄生虫发育周期，从而减少或防止寄生虫向其他人传播。预计TBV还可以防止在抗马拉里亚药物治疗过程中出现逃生突变体的扩散或针对寄生虫无性阶段的其他预防性疫苗。迄今为止，仅研究了这种类型的疫苗的四个候选分子。由于Palciparum的基因组序列于2002年10月完成，因此我们现在可以自由访问基因组数据来搜索新型疫苗候选物。但是，用于疫苗发育的瓶颈之一是恶性疟原虫外显子的含量很高，这将大大抑制使用常规方法的重组蛋白表达。我们的策略是在不使用合成基因的情况下根据恶性疟原虫的基因组序列来表达每种蛋白质的重组蛋白。为了鉴定新的TBV候选者，我们选择了190个基因，预计仅在P valciparum的配子细胞阶段表达。将这些基因克隆到质粒中，并通过基于PCR的程序制备模板进行转录，然后通过无小麦生殖细胞系统进行高吞吐量重组蛋白合成。使用这种方法，我们成功获得了120种重组蛋白。在筛选了这些重组蛋白以鉴定具有传播障碍的人血清的新型TBV候选者之后，我们将15种新型的Gametocyte抗原确定为TBV候选者。

项目成果

期刊论文数量（38）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Plasmodium ookinete-secreted proteins secreted through a common micronemal pathway are targets o blocking malaria transmission.

通过共同的微线体途径分泌的疟原虫动蛋白分泌蛋白是阻断疟疾传播的目标。

DOI：
发表时间：
2004
期刊：
Journal of Biological Chemistry 279
影响因子：
0
作者：
Toyooka;K.;et al.;Kaneko O;Yano K;Kaneko 0;Yano K;Arakawa T;Sawasaki T;Arakawa T;Sawasaki T;Arakawa T;Kaneko O;Rungruang T;Sattabongkot J;Li F;Li F
通讯作者：
Li F

Plasmmodium ookinete-secreted proteins secreted through a common micronemal pathwaay are targets of blocking malaria transmission.

通过共同的微线体途径分泌的疟原虫动蛋白分泌蛋白是阻断疟疾传播的目标。

DOI：
发表时间：
2004
期刊：
Journal of Biological Chamistry 279
影响因子：
0
作者：
Toyooka;K.;et al.;Kaneko O;Yano K;Kaneko 0;Yano K;Arakawa T;Sawasaki T;Arakawa T;Sawasaki T;Arakawa T;Kaneko O;Rungruang T;Sattabongkot J;Li F
通讯作者：
Li F

Nasal immunization with a malaria transmission-blocking vaccine candidate, Pfs25, induces complete protective immunity in mice against field isolates of Plasmodium fakiflarum.

使用阻断疟疾传播的候选疫苗 Pfs25 进行鼻腔免疫，可诱导小鼠针对现场分离的恶性疟原虫产生完全的保护性免疫力。