摘要
目的构建重组腺病毒Ad／MDC—VP1，观察Ad／MDC-VPl与DNA疫苗联合应用的免疫效果。方法构建、包装重组腺病毒Ad／MDC—VP1并检测目的蛋白的表达。BALB／c小鼠随机分为Ad／MDC—VPl、pcDNA3／MDC—VP1、pcDNA3／MDC-VP1＋Ad／MDC—VP1和PBS4组，肌肉注射免疫小鼠。用ELISA法和微量中和试验法分别检测血清柯萨奇病毒B3（CVB3）VP1 IgG和中和抗体滴度；CCK-8法检测淋巴细胞增殖活性和特异性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。结果成功构建并包装了重组腺病毒Ad／MDC-VPI，检测到目的蛋白的表达。pcDNA3／MDC．VP1＋Ad／MDC—VP1组血清CVB3VP1IgG滴度、淋巴细胞增殖指数、CTL杀伤酒生和对小鼠的保护率明显高于其他各组（P〈0．05），血清病毒滴度低于其他各组（P〈0．05）。结论Ad／MDC—VP1与DNA疫苗联合应用能显著提高小鼠细胞和体液免疫应答。
Objective To construct recombinant adenovirus Ad/MDC-VP1 and investigate its immuno-boosting effect of the mice primed with the experimental DNA vaccine against Coxsackievirus infection. Methods The recombinant adenovirus Ad/MDC-VP1 was constructed and packaged. The Western blot analysis was used to verify the target protein. BALB/c mice were divided into four groups： Ad/MDC-VP1 group, pcDNA3/MDC-VP1 group, pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost group and PBS group. The mice in each group were immunized intramuscularly. The titers of serum IgG and neutralizing antibody were tested by ELISA and trace neutralization assay, 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 Coxsackievirus, and the assay of the serum virus titers and the observation of protection efficacy against Coxsackievirus infection were carried out. Results The recombinant adenovirus Ad/MDC-VP1 was successfully constructed and the target protein was expressed. It was observed that the titers of CVB3 VP1 specific antibody, lymphocyte stimulation index, CTL cytotoxicity activities and protection rate of the pcDNA3/MDC-VPl prime-Ad/MDC-VP1 boost group were much higher than those of the rest groups（P 〈 0.05）, and the titer of serum virus was Lower after CVB3 challenged （ P 〈 0.05 ）. Cottclusion Both the cellular and humoral immune responses in mice could been significantly enhanced by the pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost strategy.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2009年第6期533-537,共5页
Chinese Journal of Microbiology and Immunology
基金
河北省科技支撑计划（072761896）
河北省医学科学研究重点课题（08258）
