副猪嗜血杆菌血清13型菌株免疫原性外膜蛋白的筛选与分析

Screening and analysis of immunogenic outer membrane proteins of Haemophilus parasuis serovar 13

为筛选副猪嗜血杆菌血清13型分离株ZJ1018具有免疫原性的外膜蛋白,本研究从该分离菌提取外膜蛋白进行双向电泳分离,并进行免疫印迹分析。共对21个蛋白点进行质谱分析,通过NCBI数据库搜索,共对应14种蛋白,包括7种外膜蛋白。其中OmpA、palA、D15为与血清5型菌株共有的免疫原性蛋白,D15蛋白在不同血清型菌株中氨基酸序列同源性不低于99.4%;蛋氨酸ABC转运体底物结合蛋白、GS60、外膜脂蛋白A属于在副猪嗜血杆菌中被筛选出的新的免疫原性外膜蛋白。本研究为副猪嗜血杆菌亚单位疫苗的开发提供了新的依据。

To search for common immunogenic proteins from different serotype of Haemophilus parasuis, immunogenic outer membrane proteins （OMPs） were screened from a serovar 13 clinical isolate through immunoproteomics approach. OMPs were extracted, and high-resolution profiles of OMPs were prepered through two-dimensional electrophoresis and immunoblot analysis. A total of 21 protein spots were identified by MALDI-TOF-MS analysis, and searched in NCBI database based on peptide mass fingerprint. It showed that 21 protein spots representing 14 proteins were identified, and 7 out of the 14 proteins were OMPs. OmpA, palA, DI5 were the three immunogenic OMPs which were identified in both serovar 13 and serovar 5 isolate, and the amino acid homology of DI5 was not less than 99.4% among strains of various serovar; Another three proteins （GS60, OmlA and methionine ABC transporter substrate-binding protein） showed to be immunogens for the first time in H.parasuis, and the potential of these proteins to be used as subunit vaccine candidates worth further investigation. The study provides new clues for development of H.parasuis subunit vaccine.