登革病毒严重威胁人类健康，中英两国面临着共同的风险。重组嵌合登革热疫苗已完成临床III期试验，显示出良好的应用前景。但是嵌合登革疫苗由反向遗传学技术拯救获得，理论上同时含两种黄病毒成分，其毒粒结构及其全蛋白的化学组成尚缺乏直接证据；同时Vero细胞生产的嵌合病毒颗粒的成熟程度上尚不明确，可能对疫苗的免疫反应产生影响。申请人与英方合作伙伴在中英合作交流项目的支持下，开展了不同条件登革病毒感染细胞的蛋白质学分析，在此基础上双方继续深化合作，结合双方优势，旨在建立基于PRM方法的靶向蛋白质组学方法，对嵌合登革热疫苗株的结构蛋白的绝对定量分析，结合动物实验明确嵌合登革疫苗颗粒成熟度对其免疫原性与抗体谱系的影响，比较分析嵌合登革疫苗及其亲本病毒诱导宿主反应的差异。本项目的完成不仅能够提供一种对嵌合登革疫苗进行绝对定量和动态监测的先进技术，而且对于深刻理解黄病毒嵌合成熟机制与宿主相互作用具有重要意义。

Dengue is now well regarded as the most prevalent and severe mosquito-borne viral disease of human beings. China and the UK are both under the risk of dengue transmission. Recently, dengue vaccine development has reached a major milestone: a live chimeric dengue vaccine candidate has shown promising results in the final stage of clinical trials. However, as a chimeric flavivirus generated by advanced DNA recombination technology, the real structure and chemical composition of chimeric dengue vaccine candidate produced in Vero cells, either in mature or immature status, is still not fully understood. Especially, the specific host cell response to the chimeric and parental viruses has been profiled. The OVERALL AIM of the proposed project is to provide advanced training in the development and application of targeted proteomics approaches for the study of a chimeric dengue vaccine candidate and the host response. The Specific Aims are to 1) Develop a targeted proteomics approach to quantitatively profile the stoichiometry of chimeric dengue vaccine virus; 2) Investigate the impact of viron maturity on antibody repertoire to each viral protein; 3) Analyse the levels of specific cellular proteins in response to infection with chimeric and the corresponding parental viruses. The project will not only be of benefit for controlling the quality of chimeric flavivirus vaccine production, but will also provide new insights into flavivirus maturation and host cell interactions. The technology developed in this proposal will be applicable to the study of other viruses or cellular systems.