流感嗜血杆菌(Hi)是社区获得性肺炎、儿童中耳炎和上感的常见病原，当前不定型Hi(NTHi)已成为主流菌型，现有Hib疫苗对其无保护性，至今尚无NTHi疫苗产品。外膜蛋白P6和PD是NTHi的优势表面抗原，在动物中具有良好保护性，但在人中免疫原性弱，而通过筛选有效的抗原表位可研制出高效安全的基因工程疫苗。本项目中，我们预测了NTHi外膜P6和PD的T-B联合表位，采用噬菌体展示、重复肽文库及ELISA技术，确定P6、PD中有效T-B联合表位；采用流式细胞及ELISA法检测不同抗原表位诱导Th1型和Th2型细胞因子的水平及其在患者中的抗体滴度，确定诱导抗感染免疫应答的Th1型T-B联合表位；制备P6和PD有效T-B联合表位的多靶点MAP候选疫苗，并通过动物实验研究其保护性。本研究为NTHi MAP疫苗产业化开发提供依据，具有较高学术价值、创新性和应用前景。

Haemophilus influenzae (Hi) is a common pathogen in children, causing community-acquired pneumoniae, otitis media and upper respiratory infections. Most common Hi infections are caused by non-typeable Hi (NTHi) strains whereas the current Hi vaccine (Hib) provides no protection because NTHi strains are noncapsulated, and there is no other vaccine for NTHi available. Outer-membrane protein P6 (OMP6) and PD are two important surface antigens in NTHi and have showed to be protective in animal models. However, they are not very immunogenic when tested in human trials. The immunogenicity of these two proteins can be improved through screening effective antigen epitopes and making multi-antigen presentation. In the current proposal, the T and B cell (T-B) common epitopes for OMP6 and PD were analyzed by software predictions. We will screen these epitopes through a phage display library and an overlap peptide library. Positive epitopes will be further validated by their ability to induce Th1 and Th2 cytokines in mice, measured by flow cytometry and ELISA. MAP will then be made from selected epitopes and tested in animal models for its protection. Successful completion of this project will pave the road for developing a MAP vaccine for NTHi and have significant implications for vaccine development for other bacteria infections.