血吸虫抗独特型抗体与细胞因子合成蛋白抗病免疫比较研究

This study demonstrated that IL-2 and IL-6 could significantly enhance the protective immunity of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum in mice，and worm reduction was elevated from 40.6% (NP30 alone) to 53.5% and 55.7% respectively. The variable region genes of light and heavy chains of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum were separated, amplified and sequenced. It was verified that a full-length VL gene was 318 bp and encoded 109 amino acids, and the full-length VH gene was 357 bp and encoded 119 amino acids. In addition, there were no original and terminal codons in the two genes, and the VH gene was an open read frame. The VL and VH gene sequences were registered by Gene Bank (accession No. AF206720 and AF282622). Then the ScFv gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum was constructed and expressed. The expressed product proved to be antigen binding activity detected with ELISA. Analysed by molecular biology computer programme, the 10 amino acids of VH CDR3 region are thought to be antigenic epitope of monoclonal antibody NP30. The 6xVH CDR3 gene of monoclonal antibody NP30 was artificially designed and constructed, then fused with IL-2 gene. The fusion protein was expressed and the protective immunity induced by fusion protein in Balb/c mice was 48.5%. This study provides a new strategy for developing glycoprotein antigenic epitope vaccine and a basis for further developing human anti-idiotypic antibody vaccine of schistosomiasis.

本项目试图制备日本血吸虫单克隆抗独特型抗体NP30与细胞因子的偶联蛋白，并应用基因工程技术构建NP30的单链抗体，将NP30 单链抗体基因与细胞因子基因拼接，表达融合蛋白。观察偶联蛋白和融合蛋白对日本血吸虫感染的保护率，并研究比较二者对血吸虫娣⒂⒋瞥娌选⒊媛雅咛シ⒂⒊媛讶庋恐仔纬珊拖宋髌诘挠跋臁Ｎ夜娌∫呙缪兄瓶匾惶跣滤悸贰