基于流感病毒HA柄部和M2e融合基因的DNA疫苗  

An Influenza DNA Vaccine Based on a Fusion Gene of HA Stalk and M2e Domains

作　　者：贺彪[1] 常海艳[1] 李小曼[1] 陈则[1] 方芳[1] 

机构地区：[1]湖南师范大学生命科学学院,中国湖南长沙410081

出　　处：《生命科学研究》2013年第4期325-330,341,共7页Life Science Research

基　　金：湖南省高校创新平台开放基金资助项目(11K040);湖南省科技厅基金资助项目(2011SK3109)

摘　　要：由于流感病毒容易突变,流感通用疫苗的研究势在必行.流感病毒血凝素(HA)柄部和基质蛋白2的胞外域(M2e)都是流感病毒通用疫苗的重要候选靶点.通过重叠PCR的方法用A/PR/8/34(H1N1)(简称PR8)流感病毒的M2e或者4个甘氨酸取代HA的头部,分别获得HAM2e和HA4G,然后将两种重组基因插入真核表达载体pCAGGS-P7中,制得pHAM2e和pHA4G两种DNA疫苗.通过电击免疫的方法对小鼠分别免疫pHAM2e和pHA4G,免疫3次,每次免疫间隔2周.第3次免疫2周后用5LD50的同源流感病毒感染小鼠.结果表明HAM2e组和HA4G组的小鼠均产生了特异性抗体,HAM2e组比HA4G组具有更好的抗PR8流感病毒的能力,这提示可以用M2e替换HA头部用于疫苗研发.Universal influenza vaccines have been extensively studied due to the antigenic variation of influenza virus. The haemagglutinin （HA） stalk region and the matrix protein 2 （M2） extracellular domain （M2e） are two important candidates in developing universal vaccines. In this study, the head region of influenza virus A/PR/8/34 （H1N1） HA was replaced by M2e and 4 glycines, respectively, through overlapping PCR. The plasmids pHAM2e and pHA4G were constructed by inserting recombinant genes HAM2e and HA4G into the expression vector pCAGGS-P-/, respectively. Mice were immunized by the constructed plasmid using electroporation 3 times at a two-week interval. Two weeks after the final immunization, mice were challenged with a lethal dose （5LDs0） of PR8 virus. Specific antibodies can be detected in mice immunized with either pHAM2e or pHA4G, pHAM2e provided better protection than pHA4G against viral challenge. These results demonstrated the feasibility of replacing the head domain of HA by M2e for future development of influenza vaccines.

关 键 词：流感 血凝素 基质蛋白2 通用疫苗 

分 类 号：R183.3[医药卫生—流行病学]