摘要
为评价犬恶丝虫硫氧还蛋白过氧化物酶(TPx)真核表达质粒的免疫原性,本实验利用RT-PCR方法扩增TPx基因,将其克隆于真核表达载体pVAX1中构建重组质粒pVAX1-TPx,并对其进行体外表达鉴定及通过特异性抗体水平及相关的免疫因子的检测,评价其在体内诱导的免疫反应。实验结果表明,将pVAX1-TPx转染于Cos7细胞中能够正确表达TPx,其分子量约为28 ku,并被阳性血清所识别。将pVAX1-TPx免疫BALB/c小鼠并采用ELISA检测结果显示,重组质粒免疫组的外周血中抗体、Th2细胞分泌的IL4及IL13细胞因子水平均显著高于空质粒及空白对照组(p<0.05);但Th1分泌的IFN-γ及IL2水平差异不显著。此外,淋巴细胞增殖试验结果也表明,pVAX1-TPx免疫组显著高于其他两个对照组(p<0.05)。实验数据表明pVAX1-TPx免疫可以有效诱导特异性的体液和细胞免疫。
To evaluate the immunogenicity of a DNA vaccine encoding thioredoxin peroxidase （TPx） of Dirofilaria immitis, the TPx gene was amplified by RT-PCR and cloned into expression vector of pVAX1 to construct the pVAX1-TPx as DNA vaccine. The recombinant TPx was expressed in Cos7 cell line transfected with pVAX1-TPx and recognized by the specific TPx antibody prepared in mice with the prokaryotic expressed recombinant TPx. Furthermore, the mice were immunized with pVAX1-TPx via intramuscular injection and the antibody levels against TPx in mice immunized group were significantly higher than that in pVAX1 or blank control groups （p〈0.05）, and the expression levels of IL4 and ILl3 were also significantly higher in immunized group than that in the control groups, respectively （/9〈0.05）. In conclusion, the TPx was capable of stimulating the spleen ceils proliferation, indicating that the TPx possessed promising immunogenicity agains D.immitis.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2013年第4期325-329,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
中央高校基本科研业务费专项资金(XDJK2009B001)
关键词
犬恶丝虫
TPx
表达质粒
免疫原性
Dirofilaria immitis
TPx gene
DNA vaccine
immunogenicity
