Scanning the P. falciparum proteome for vaccine antigens

描述（由申请人提供）：来自感染性微生物的大量基因组数据提供了丰富的信息，现在可用于确定亚单位疫苗开发的抗原靶点。本文以恶性疟原虫为模型，提出了一种利用生物信息学和高通量蛋白质组学技术从大而复杂的基因组中快速识别靶抗原的策略。接触完整的寄生虫可诱发对疟疾的保护性免疫。然而，在疟原虫的生命周期中有超过5000种蛋白表达，而介导机体免疫诱导的保护性免疫的蛋白抗原在很大程度上是未知的。迄今为止，只确定了少数潜在目标。目前正在开发的亚单位疫苗基于单一或少数抗原，因此可能引起的反应范围过窄，既不能提供最佳保护，也不能对遗传多样性背景提供保护。PCR Express是一种高通量蛋白质表达平台技术，可用于从任何测序的微生物中快速生成完整的蛋白质组。PCR Express允许数千种不同的基因被克隆、表达并打印到蛋白质微阵列芯片上，每周的速度超过300种蛋白质。该芯片可用于扫描接种疫苗或感染的动物和人类的血清以及确定的免疫优势抗原。然后，这些宝贵的知识可以立即用于拟议研究的其他领域：1)用于测量疫苗接种者疟疾免疫力的诊断工具；2)开发用于评估近期疟疾暴露/感染的简单、快速和敏感的免疫工具；3)研制治疗性抗疟疾中和性单克隆人抗体；4)开发安全有效的候选亚单位疟疾疫苗；5)准确评估替代疟疾疫苗引起的免疫反应的质量。特别重要的是，迫切需要一种疟疾疫苗，从这一申请中产生的数据将用于确定可能用于抗疟疾DNA或蛋白质亚基疫苗的保护性抗原候选物。

DESCRIPTION (provided by applicant): The abundance of genomic data from infectious microorganisms provides a wealth of information that may now be used to identify antigen targets for subunit vaccine development. Here we propose a strategy employing bioinformatics together with high throughput proteomics to rapidly identify target antigens from large and complex genomes, using Plasmodium falciparum as a model. Protective immunity to malaria can be induced by exposure to intact parasite. However, more than 5000 proteins are expressed during the life cycle of the Plasmodium spp. parasite, and the protein antigens mediating the protective immunity induced by whole organism vaccination are largely unknown. Only a small number of potential targets have been identified to date. Subunit vaccines currently in development are based on a single or few antigens and may therefore elicit too narrow a breadth of response, providing neither optimal protection nor protection on genetically diverse backgrounds. A high throughput protein _expression platform technology, called PCR Express, has been developed which can be used to rapidly generate the complete proteome from any sequenced microorganism. PCR Express allows thousands of different genes to be cloned, expressed and printed onto protein microarray chips, at a rate of more than 300 proteins per week. The chips can be used to scan serum from vaccinated or infected animals and humans and the immunodominant antigens identified. This valuable knowledge can then be immediately parlayed into additional areas of proposed research: 1) diagnostic tools for measuring malaria immunity in vaccinees; 2) development of simple, rapid, and sensitive immunological tools for assessing recent malaria exposure/ infection; 3) development of therapeutic anti-malaria neutralizing monoclonal human antibodies; 4) development of safe and effective candidate subunit malaria vaccines; and 5) accurate assessment of the quality of the immune responses elicited by alternative malaria vaccines. Of particular importance, there is a serious need for a malaria vaccine and the data generated from this application will be used to identify likely protective antigen candidates for a DNA or protein subunit vaccine against malaria.