rAd/MDC-VP1初免pcDNA3/MDC-VP1加强策略抗CVB3的免疫效果研究  

Study on combined immunization of rAd/MDC-VP1 and pcDNA3/MDC-VP1 against Coxsackievirus B3 challenge in mice

作　　者：闫立景[1] 高志云[2] 李剑[2] 蓝佳明[2] 金玉怀[2] 谢立新[2] 揣侠[2] 王永祥[2] 

机构地区：[1]江南大学医院检验科,江苏无锡214122 [2]河北医科大学病原生物学教研室,河北石家庄050017

出　　处：《细胞与分子免疫学杂志》2012年第3期228-231,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：河北省医学科学研究重点课题(20090049;08265);河北省科技支撑计划(072761896)

摘　　要：目的:应用柯萨奇病毒B3(CVB3)腺病毒载体疫苗rAd/MDC-VP1初免,核酸疫苗pcDNA3/MDC-VP1加强免疫的策略免疫小鼠,观察其免疫效果。方法:BALB/c小鼠随机分为A～D 4组,分别肌肉注射PBS、rAd/MDC-VP1、pcD-NA3/MDC-VP1、rAd/MDC-VP1+pcDNA3/MDC-VP1,用ELISA和微量中和试验法分别检测CVB3 VP1特异性IgG和中和抗体滴度,CCK-8法检测脾淋巴细胞增殖活性和特异性CTL杀伤活性;用致死量的CVB3攻击小鼠后,检测小鼠血中病毒滴度并观察动物的存活情况。结果:D组CVB3 IgG、非特异性淋巴细胞增殖活性及特异性CTL杀伤活性明显高于其他各组(P<0.05);CVB3攻击后,D组小鼠血中病毒滴度较其他各组显著降低,生存率为41.67%,明显高于其他各组(P<0.05)。结论:rAd/MDC-VP1初免pcDNA3/MDC-VP1加强的免疫策略能显著提高小鼠细胞和体液免疫水平,提高致死量病毒攻击后的保护率。AIM： To immunize the mice using the rAd/MDC-VP1 prime-pcDNA3/MDC-VP1 boost strategy and observe its immunological effect against Coxsackievirus B3（CVB3）.METHODS： BALB/c mice were randomly divided into four groups： PBS group,rAd/MDC-VP1 group,pcDNA3/MDC-VP1 group and rAd/MDC-VP1 prime-pcDNA3/MDC-VP1 boost group.Mice in each group were immunized intramuscularly.The titers of serum IgG and neutralizing antibody were tested by ELISA and trace neutralization assays respectively.The Lymphocytes proliferation activity and specific CTL cytotoxic activity were tested by CCK-8 assay.The mice in each group were challenged with lethal dose of CVB3,and the serum virus titer was assayed and the protection efficacy against Coxsackievirus infection was observed.RESULTS： It was observed that the titers of CVB3 VP1 specific IgG and neutralizing antibody,non-specific lymphocytic proliferation activity and specific lymphocytic CTL cytotoxic activity of the rAd/MDC-VP1 prime-pcDNA3/MDC-VP1 boost group were much higher than those of the rest groups（P〈0.05）,what＇s more,after CVB3 challenged,the serum virus titer of this group was lower and the protection rate（41.67%） was higher（P〈0.05）.CONCLUSION： Both the cellular and humoral immune responses in mice could be significantly enhanced by the rAd/MDC-VP1 prime-pcDNA3/MDC-VP1 boost strategy and the protection rate after challenged by lethal dose of virus could be increased.

关 键 词：柯萨奇病毒B3 初免-加强免疫策略 小鼠 

分 类 号：R373.2[医药卫生—病原生物学]