Large Scale systematic priorization of Plasmodium vivax blood stage vaccine antigens

摘要 尽管最近在减少撒哈拉以南非洲的严重疾病方面取得了进展，但疟疾仍然 全球最大的公共卫生负担之一，也是#年儿童死亡的主要原因 低收入和中等收入国家。迫切需要一种有效的疟疾疫苗 因为所有已公开释放的抗疟疾药物都出现了抗药性 到目前为止。针对恶性疟原虫的疫苗，是非洲疟疾的主要原因，已经 得到了广泛的支持，并显示出令人鼓舞的迹象，但疫苗研究 间日疟原虫是非洲以外疟疾的主要原因，到目前为止， 有限的。这项提议将利用恶性疟原虫疫苗开发的经验教训，用于 太长时间专注于有限数量的候选人，没有充分利用深度 现有的基因组序列信息。中心目标是实现第一个 间日疟原虫血液期抗原的全面反向疫苗学评估， 将先进的基因组和细胞技术与体外表型分析相结合。一个 所建立的真核蛋白表达系统将用于表达&gt；200P. 间日疟原虫血液期疫苗候选疫苗，将根据基因表达进行选择， 生物物理特征和基因组多样性标准。所有表达式构造都将是 免费提供给研究界，以帮助全球间日疟原虫疫苗和生物学 研究努力。将对表达的蛋白进行抗体结合筛选，以确认 正确的折叠，并用于红细胞和受体结合分析，以确定靶点 进一步研究。将针对100个靶点制备多克隆抗体，并用于体外P。 间日疟原虫侵袭表型分析，以确定阻止寄生虫入侵或 成长。来自南美和东南亚的间日疟原虫将在这次会议上使用 筛选阶段，在目标优先排序的最早阶段引入抗原多样性， 恶性疟原虫疫苗开发的另一个重要教训是针对特定毒株的疫苗 抑制反应让几个有希望的候选人脱轨。抗体提供了交叉- 将联合进行菌株抑制试验，以鉴定&gt；4高效和协同作用 未来开发和潜在临床测试的候选者。这项提议将从根本上 改变间日疟原虫疫苗开发的规模和步伐，并将产生 对疟疾研究界具有广泛的实用价值。

Summary Despite recent progress in reducing severe disease in sub-Saharan Africa, malaria remains one of the largest global public health burdens and a leading cause of mortality in children in low and middle income countries. There is an urgent need for an effective malaria vaccine because resistance has emerged to every antimalarial drug that has been publicly released to date. Vaccines against Plasmodium falciparum, the major cause of malaria in Africa, have received extensive support and are showing encouraging signs, but vaccine research for Plasmodium vivax, the major cause of malaria outside of Africa, has to date been extremely limited. This proposal will use the lessons from P. falciparum vaccine development, which for too long focussed on a limited number of candidates and did not make use of the full depth of available genomic sequence information. The central objective is to carry out the first comprehensive reverse vaccinology assessment of P. vivax blood stage antigens, combining advanced genomic and cellular techniques with ex vivo phenotyping assays. An established eukaryotic protein expression system will be used to express a library of >200 P. vivax blood stage vaccine candidates, which will be selected based on gene expression, biophysical characteristics and genomic diversity criteria. All expression constructs will be made freely available to the research community to aid global P. vivax vaccine and biology research efforts. The expressed proteins will be screened for antibody binding to confirm correct folding, and used in erythrocyte and receptor binding assays to prioritise targets for further study. Polyclonal antibodies will be raised against 100 targets and used in ex vivo P. vivax invasion phenotyping assays, to identify antibodies that block parasite invasion or growth. P. vivax parasites from both South America and Southeast Asia will be used at this stage of screening, introducing antigenic diversity at the earliest stage of target prioritization, another important lesson from P. falciparum vaccine development where strain-specific inhibitory responses derailed several promising candidates. Antibodies that provide cross- strain inhibition will be tested in combination to identify >4 highly effective and synergistic candidates for future development and potential clinical testing. The proposal will radically change the scale and pace of P. vivax vaccine development, and will produce resources of broad utility to the malaria research community.