CVB3腺病毒载体疫苗Ad/VP1的构建及免疫小鼠的效果  

Construction of the Recombinant Adenovirus Expressing the CVB3 VP1 protein and study on its immunological effects in mice

作　　者：李嘉[1] 李剑[1] 温婵[1] 羡鲜[1] 张永红[1] 金玉怀[1] 王永祥[1] 

机构地区：[1]河北医科大学病原生物学教研室,河北石家庄050017

出　　处：《基础医学与临床》2011年第11期1251-1255,共5页Basic and Clinical Medicine

基　　金：河北省科学技术研究与发展计划(08276101D-93)

摘　　要：目的构建柯萨奇病毒B3(CVB3)VP1基因重组腺病毒疫苗Ad/VP1,并观察其对小鼠的免疫效果。方法利用AdEasy-1系统构建、包装重组腺病毒Ad/VP1,Western blot法检测目的蛋白的表达。BALB/c小鼠随机分为Ad/VP1、Ad和PBS 3组,肌肉注射免疫,共2次,间隔16 d。用ELISA法和微量中和试验法分别检测血清CVB3 VP1IgG和中和抗体滴度;CCK-8法检测特异性CTL杀伤活性;用致死量CVB3攻击小鼠后,检测血中病毒滴度并观察小鼠的存活率。结果成功构建、包装了重组腺病毒Ad/VP1,并在293细胞中检测到VP1蛋白的表达。末次免疫小鼠后,Ad/VP1组血清CVB3 VP1 IgG滴度为168.3±1.4、中和抗体水平为31.8±1.4,均明显高于PBS和Ad对照组(P<0.01),CTL杀伤活性和对小鼠的保护率也高于对照组(P<0.05),血清病毒滴度低于两对照组(P<0.05)。结论重组腺病毒疫苗Ad/VP1能显著提高小鼠的细胞和体液免疫应答及免疫保护作用。Objective To construct a recombinant adenovirus Ad/VP1 and to investigate the immune effect against coxsackievirus infection in mice.Methods The recombinant adenovirus Ad/VP1 was constructed and packaged.The target protein was verified by Western blot analysis.BALB/c mice were divided into three groups： Ad/VP1 group,Ad group and PBS group.The mice in each group were immunized by intramuscular injection.The titer of sera IgG and neutralizing antibody was detected by ELISA and trace neutralization assay.The specific CTL cytotoxic activity was detected by CCK-8 assay.The mice in each group were challenged with lethal dose of coxsackievirus,the virus load in peripheral blood was titrated and the protection efficacy against coxsackievirus infection was observed.Results The recombinant adenovirus Ad/VP1 was successfully constructed and target protein was expressed.The titers of CVB3 VP1 specific antibody and neutralizing antibody was much higher than those in the other groups（P0.01）,CTL cytotoxicity activities and protection rate of the Ad/VP1 group were also much higher than the other groups（P0.05）,and the virus load was lower after CVB3 challenged（P0.05）.ConclusionBoth the celluar and humoral immune responses and the protection rate in mice could been significantly enhancedby the Ad/VP1.

关 键 词：柯萨奇病毒B3 腺病毒载体疫苗 小鼠 

分 类 号：R392.12[医药卫生—免疫学]