BACKGROUND Immunization with attenuated malaria sporozoites protects humans from experimental malaria challenge by mosquito bite. Protection in humans is strongly correlated with the production of T cells targeting a heterogeneous population of pre-erythrocyte antigen proteoforms, including liver stage antigens. Currently, few T cell epitopes derived from Plasmodium falciparum, the major aetiologic agent of malaria in humans are known. METHODS In this study both in vitro and in vivo malaria liver stage models were used to sequence host and pathogen proteoforms. Proteoforms from these diverse models were subjected to mild acid elution (of soluble forms), multi-dimensional fractionation, tandem mass spectrometry, and top-down bioinformatics analysis to identify proteoforms in their intact state. RESULTS These results identify a group of host and malaria liver stage proteoforms that meet a 5% false discovery rate threshold. CONCLUSIONS This work provides proof-of-concept for the validity of this mass spectrometry/bioinformatic approach for future studies seeking to reveal malaria liver stage antigens towards vaccine development.

中文翻译：

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从肝阶段模型鉴定恶性疟原虫的蛋白形式。

背景技术减弱的疟疾子孢子的免疫保护人类免受蚊子叮咬的实验性疟疾攻击。对人类的保护与靶向异种前红细胞抗原蛋白形式（包括肝阶段抗原）的T细胞的产生密切相关。目前，很少有人知道源自疟疾的主要病原体恶性疟原虫的T细胞表位。方法在本研究中，使用体外和体内疟疾肝阶段模型对宿主和病原体蛋白形式进行测序。对来自这些不同模型的蛋白形式进行温和的酸洗脱（可溶形式），多维分馏，串联质谱和自上而下的生物信息学分析，以鉴定其完整状态的蛋白形式。结果这些结果确定了一组符合5％错误发现率阈值的宿主和疟疾肝阶段蛋白形式。结论这项工作为该质谱/生物信息学方法的有效性提供了概念验证，可用于未来的研究，以揭示疟疾肝阶段抗原用于疫苗开发。