权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Comprehensive screening and identification of the novel malaria transmission-blocking vaccine candidate antigens

新型疟疾传播阻断疫苗候选抗原的综合筛选和鉴定

基本信息

批准号：
18390129
负责人：
TSUBOI Takafumi
金额：
$ 10.49万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390129/
关键词：
malaria vaccine infectious diseases biotechnology genome International scientist exchange Thailand

项目摘要

Malaria transmission-blocking vaccines (TBVs) prevent the transmission of malaria by inducing antibodies against antigens specifically expressed on the sexual stage parasites. Since well-characterized TBV candidates are only four, it would be necessary to identify novel TBV candidates for a successful TBV development. In order to identify the novel TBV candidates, we searched a combined dataset from genome and transcriptome databases and selected 192 genes, which are expected to be expressed only in gametocyte stage of P. falciparum. These genes were cloned into plasmids and 170 of the template cDNA clones 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 148 recombinant proteins. After the screening of these recombinant proteins to identify novel TBV candidates with malaria patient sera harboring parasite transmission-blocking antibodies, we have identified 27 novel antigens. After subclone these candidate genes into pEU plasmid for the large-scale expression using the wheat germ cell-free system, we have expressed and affinity purified 22 targets. In order to obtain antisera, we immunized these proteins into mice. After the confirmation of the immunoreactivity of these antisera against parasites, we examined the transmission-blocking efficacy of the sera. Finally we identified the two novel parasite antigens, which induced transmission-blocking antibodies in mice. Accordingly, this approach will be useful for the novel transmission-blocking vaccine candidate discovery.

疟疾传播阻断疫苗（TBVs）通过诱导针对性期寄生虫特异性表达的抗原的抗体来预防疟疾的传播。由于特征明确的TBV候选只有四个，因此有必要确定新的TBV候选，以便成功地开发TBV。为了鉴定新的TBV候选者，我们从基因组和转录组数据库中搜索组合数据集，并选择了192个基因，这些基因预计仅在恶性疟原虫配子体阶段表达。将这些基因克隆到质粒中，并通过基于PCR的程序制备170个模板cDNA克隆用于转录，然后通过麦胚无细胞系统进行高通量重组蛋白合成。利用这种方法，我们成功地获得了148个重组蛋白。在筛选这些重组蛋白以鉴定具有寄生虫传播阻断抗体的疟疾患者血清的新型TBV候选物之后，我们鉴定了27种新抗原。将这些候选基因亚克隆到pEU质粒中，利用麦胚无细胞系统进行大规模表达，共表达并亲和纯化了22个目的基因。为了获得抗血清，我们将这些蛋白免疫小鼠。在确认这些抗寄生虫的抗血清的免疫反应性后，我们检查了血清的传播阻断效力。最后，我们鉴定了两种新的寄生虫抗原，它们在小鼠中诱导了传播阻断抗体。因此，这种方法将是有用的新的传播阻断疫苗候选人的发现。