【作者】 徐红卫；

【导师】 王天厚；

【作者基本信息】 华东师范大学 ， 生态学， 2011， 硕士

【摘要】 为了制备和筛选针对甲型和乙型流感病毒具有广谱和强抗病毒活力的抗体和多肽,我们应用杂交瘤技术分别制备了针对甲型流感病毒M2和乙型流感病毒BM2离子通道的单克隆抗体,同时应用噬菌体展示技术筛选出与乙型流感病毒M2离子通道膜外区高特异结合的噬菌体多肽,并分别从细胞水平上研究了它们的抗病毒活性。结果分别筛选到针对甲型和乙型流感病毒M2膜外区N末端8个保守氨基酸序列的单克隆抗体M2e8-7和BM2e-2,其中M2e8-7抗体高特异性结合已知21种甲型流感病毒M2突变型中的19种,是已报道的M2单克隆抗体中识别病毒种类最多的一株抗体。病毒中和实验显示M2e8-7能有效抑制所识别的不同甲型流感病毒突变型在MDCK细胞里的复制。与此类似,BM2e-2抗体有效抑制了不同乙型流感病毒株在MDCK细胞里的复制。此外,还筛选到了能与乙型流感病毒M2膜外区8个保守氨基酸序列高度特异结合的短肽VSFTPSF,其抗病毒活性还有待进一步研究。因此,M2离子通道的独特结构对于抗病毒感染具有重要的作用,本研究所筛选到的针对甲型和乙型流感病毒特异的单克隆抗体和短肽为临床检测和抗病毒药物的开发提供了重要的理论依据。更多还原

【Abstract】 To investigate the broad spectrum antiviral activities in vitro, we have prepared and characterized the monoclonal antibodies against M2 protein channel of influenza A and B viruses. Not only have we applied the hybridoma technique to get the monoclonal antibodies against M2 protein of influenza A and B viruses, but also screened the phage displayed peptide specifically binding to BM2. We characterized the M2e8-7and BM2e-2 recognizing the N terminus eight highly conserved epitomes respectively. The mAb M2e8-7 exhibited high affinity reacted with 19 of the current known 21 influenza A M2e variants. A cytopathic assay showed that the mAb M2e8-7 potently inhibited the replication of variety influenza A virus. Seemly, the monoclonal antibody BM2e-2 potently protected MDCK cells from homologous, but not heterologous, virus infections. Besides, specific peptide sequence VSFTPSF secreting anti-BM2 mAbs were obtained and the mechanism of the inhibition was further discussed. These results indicate that antibody targeting the M2 proton channel is a promising therapeutic candidate for treating influenza virus infections, and that brought new insight in developing vaccine.更多还原