耐药性甲型流感病毒H3N2神经氨酸酶免疫原性的研究

摘要

To determine whether the immunogenicity of the neuraminidase (NA) in drug-resistance influenza virus H3N2 changed by comparison with the wild type NA,two key glycopeotein,hemagglutinin (HA) and NA,of wild type influenza virus H3N2 were obtained by prokaryotic expression in E coli.The immunogenicity of the neuraminidase was analyzed by Western blotting,ELISA detection and neutralizing antibody assay of H3N2 wild type and mutation types against HA and NA immuned serum.The results indicated that the wild type and E119D type exhibit high immunoreactivity against HA and NA immuned serum while much lower in the E119D+I222L type and I222L type.It first determined that the immunogenicity of the neuraminidase in drug-resistance influenza virus H3N2 had significantly difference from the wild type and I222L type,which allowed H3N2 to escape the immune attack and acquire the drug-resistance activity.This would provide a solid base for the development of the modern H3N2 diagnostic reagents and vaccine in the near future.%为了确定耐药性甲型流感病毒H3N2神经氨酸酶的抗原性质较野生型甲型流感病毒是否发生改变,利用原核表达方法在大肠杆菌中大量表达并纯化野生型甲型流感病毒H3N2的红细胞凝集素(HA)和神经氨酸酶(NA)；取HA和NA免疫后的小鼠血清分别对野生型和突变型甲型流感病毒H3N2进行Westernblotting、ELISA以及中和抗体检测.结果表明,野生型甲型流感病毒以及第119位氨基酸单点突变型甲型流感病毒对HA和NA免疫后的小鼠血清都有很好的免疫反应性,而第222位氨基酸单点突变和双点耐药突变型甲型流感病毒对免疫血清的免疫反应性要低很多.该研究首次确定了甲型流感病毒H3N2的野毒株与耐药突变株的神经氨酸酶的抗原性质存在很大的差异,进一步表明耐药突变株病毒除了具有抗药物活性之外也具有逃逸机体所产生的抗体攻击的能力,可为将来新一代甲型流感病毒H3N2诊断试剂和疫苗的开发等进一步的研究提供重要参考.