Guo Jianqiang, Yao Lihong, Chen Aijun, Liu Xiaoyu, Fu Jinqi, Xu Pengwei, Zhang Zhiqing
State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Sheng Wu Gong Cheng Xue Bao. 2011 Jun;27(6):876-83.
In order to evaluate the response to vector-expressed M1 and HA genes of influenza virus in mice, we prepared recombinant plasmid pStar-M1/HA and recombinant adenovirus Ad-M1/HA containing both the full-length matrix protein 1(M1) and hemagglutinin (HA) genes of human H5N1 influenza virus strain A/Anhui/1/2005. We then combined the DNA vaccine and adenoviral vaccine in immunization of BALB/c mice with a prime-boost regime. We immunized the mice with DNA vaccine at day 0 and 28 and with recombinant adenoviral vaccines at day 14 and 42. We took blood samples before each injection and 14 days after the final injection for detection of humoral immune responses. At day 56, we sacrificed the mice and collected splenocytes for detection of cellular immune responses. ELISA and hemagglutination inhibition (HI) assay showed that specific IgG Abs against H5N1 influenza virus was induced in serum of the immunized mice. ELISPOT results confirmed that the specific cellular immune responses were successfully induced against the M1 and HA proteins of H5N1 influenza virus. This study provides new strategy for development of novel influenza vaccines.
为了评估小鼠对流感病毒载体表达的M1和HA基因的反应，我们制备了重组质粒pStar-M1/HA和重组腺病毒Ad-M1/HA，它们包含人H5N1流感病毒株A/Anhui/1/2005的全长基质蛋白1（M1）和血凝素（HA）基因。然后，我们采用初免-加强免疫方案，将DNA疫苗和腺病毒疫苗联合用于BALB/c小鼠的免疫。我们在第0天和第28天用DNA疫苗免疫小鼠，并在第14天和第42天用重组腺病毒疫苗免疫。我们在每次注射前和最后一次注射后14天采集血样，以检测体液免疫反应。在第56天，我们处死小鼠并收集脾细胞，以检测细胞免疫反应。ELISA和血凝抑制（HI）试验表明，免疫小鼠血清中诱导产生了针对H5N1流感病毒的特异性IgG抗体。ELISPOT结果证实，成功诱导了针对H5N1流感病毒M1和HA蛋白的特异性细胞免疫反应。本研究为新型流感疫苗的开发提供了新策略。
