含CpG序列PRRSV ORF5基因真核表达质粒在小鼠的细胞免疫应答

Cellular immune response to eukaryotic expression vector containing PRRSV ORF5 and CpG motifs

为了提高猪繁殖与呼吸综合征病毒(PRRSV)基因免疫的效果,构建含CpG基序和PRRSV ORF5的真核重组表达质粒CpG-pVAX1-ORF5。经酶切鉴定和序列测定,将重组质粒转染COS-7细胞,经间接免疫荧光试验证实CpG-pVAX1-ORF5可表达PRRSV GP5蛋白。免疫SPF小鼠,检测鼠的脾T淋巴细胞亚群数量(CD4+和CD8+)以及淋巴细胞增殖反应(MTT法)水平。结果表明本试验构建的含CpG基序真核重组表达质粒CpG-pVAX1-ORF5能诱导小鼠产生针对PRRSV的细胞免疫。

To improve the gene immunization against porcine reproductive and respiratory syndrome virus （PRRSV）, the eukaryotic expression vector CpG-pVAX1- ORF5 containing PRRSV ORF5 gene coding region and CpG motif we constructed After identified by enzyme analysis and nucleotide sequencing test, the recombinant expression vector plasmid CpG-pVAX1-ORF5 was transfected into CO8-7 cells. The transient expression of PRRSV ORF5 nueleoeapsid protein was detected by indirect immunofluorescence assay（IFA）. The recombinant expression plasmid was inoculated into SPF mice. The number of CD4 ＋, CD8 ＋ and the lymphocyte proliferation test （MTT） in peripheral blood of the vaccinated mice were used to detect. The results showed that ehd recombinant plasmid CpG-pVAX1- ORF5 could induce higher cellular immune response.