共表达H5N1流感病毒M1和HA基因的DNA疫苗与腺病毒载体疫苗的小鼠免疫评价(英文)

Evaluation of the immune response to recombinant DNA vaccine and adenoviral vaccine co-expressing the M1 and HA genes of H5N1 influenza virus in mice

为评价在小鼠体内表达流感病毒M1和HA基因诱导的免疫反应,制备共表达H5N1亚型禽流感病毒(A/Anhui/1/2005)全长基质蛋白1(M1)基因和血凝素(HA)基因的重组DNA疫苗pStar-M1/HA和重组腺病毒载体疫苗Ad-M1/HA,将其按初免-加强程序免疫BALB/c小鼠,共免疫4次,每次间隔14 d。第1、3次用DNA疫苗,第2、4次用重组腺病毒载体疫苗,每次免疫前及末次免疫后14 d采集小鼠血清用于检测体液免疫应答,末次免疫后14 d采集小鼠脾淋巴细胞用于检测细胞免疫应答。血凝抑制(HI)实验和ELISA实验检测到免疫后小鼠血清中抗H5N1亚型流感病毒特异性IgG抗体。ELISPOT实验检测到免疫后小鼠针对M1和HA的特异性细胞免疫应答。共表达H5N1流感病毒M1和HA基因的DNA疫苗与腺病毒载体疫苗在小鼠体内的免疫学评价,为研究开发新型重组流感疫苗奠定了基础。

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.