Zheng Liyun, Wang Fuyan, Yang Zhongdong, Chen Jianjun, Chang Haiyan, Chen Ze
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
BMC Infect Dis. 2009 Feb 12;9:17. doi: 10.1186/1471-2334-9-17.
Developing vaccines for the prevention of human infection by H5N1 influenza viruses is an urgent task. DNA vaccines are a novel alternative to conventional vaccines and should contribute to the prophylaxis of emerging H5N1 virus. In this study, we assessed whether a single immunization with plasmid DNA expressing H5N1 hemagglutinin (HA) could provide early protection against lethal challenge in a mouse model.
Mice were immunized once with HA DNA at 3, 5, 7 days before a lethal challenge. The survival rate, virus titer in the lungs and change of body weight were assayed to evaluate the protective abilities of the vaccine. To test the humoral immune response induced by HA DNA, serum samples were collected through the eye canthus of mice on various days after immunization and examined for specific antibodies by ELISA and an HI assay. Splenocytes were isolated after the immunization to determine the antigen-specific T-cell response by the ELISPOT assay.
Challenge experiments revealed that a single immunization of H5N1 virus HA DNA is effective in early protection against lethal homologous virus. Immunological analysis showed that an antigen-specific antibody and T-cell response could be elicited in mice shortly after the immunization. The protective abilities were correlated with the amount of injected DNA and the length of time after vaccination.
A single immunization of 100 mug H5 HA DNA vaccine combined with electroporation was able to provide early protection in mice against homologous virus infection.
研发预防H5N1流感病毒人间感染的疫苗是一项紧迫任务。DNA疫苗是传统疫苗的一种新型替代物,应有助于预防新出现的H5N1病毒。在本研究中,我们评估了用表达H5N1血凝素(HA)的质粒DNA进行单次免疫是否能在小鼠模型中提供针对致死性攻击的早期保护。
在致死性攻击前3、5、7天用HA DNA对小鼠进行一次免疫。测定存活率、肺内病毒滴度和体重变化以评估疫苗的保护能力。为检测HA DNA诱导的体液免疫反应,在免疫后不同天数通过小鼠眼角收集血清样本,并通过ELISA和血凝抑制试验检测特异性抗体。免疫后分离脾细胞,通过ELISPOT试验确定抗原特异性T细胞反应。
攻毒实验表明,单次免疫H5N1病毒HA DNA可有效提供针对致死性同源病毒的早期保护。免疫分析显示,免疫后不久小鼠体内可引发抗原特异性抗体和T细胞反应。保护能力与注射的DNA量和接种后的时间长度相关。
单次免疫100μg H5 HA DNA疫苗并结合电穿孔能够在小鼠中提供针对同源病毒感染的早期保护。