【摘要】 为探讨鸡IL-18基因真核表达质粒pcDNA-chIL-18在新城疫病毒(NDV)F基因疫苗免疫中的作用,克隆了新城疫病毒标准株F48E9的F基因,构建F基因重组真核表达质粒pcDNA-F;将11日龄试验鸡分为4组,每组10只,14日龄一免,28日龄二免,分别肌注pcDNA-F、pcDNA-F+pcDNA-chIL-18、新城疫传统疫苗(14日龄NDV弱毒疫苗首免,28日龄NDV油苗二免)、生理盐水,一免疫后第7、14、21、28 d均采血进行ND抗体(ELISA)检测和T淋巴细胞转化(MTT)试验;第29 d均肌注50LD50NDV强毒F48E9株.结果显示:免疫保护效率,生理盐水组0/10,pcDNA-F免疫组3/10,pcDNA-F+pcDNA-chIL-18免疫组5/10,新城疫传统疫苗免疫组10/10;ND抗体水平,pcDNA-F免疫组与pcDNA-F+pcDNA-chIL-18免疫组无差异(P>0.05),均显著低于传统疫苗免疫组(P<0.05);T细胞转化水平,pcDNA-F+pcDNA-chIL-18免疫组明显高于pcDNA-F免疫组(P<0.05),pcDNA-F免疫组与传统疫苗免疫组无差异(P>0.05).试验结果综合显示pcDNA-F免疫试验鸡可产生一定程度的免疫保护;pcDNA-chIL-18对pcD-NA-F具有一定的免疫增强作用.更多还原

【Abstract】 To study the effect of immunization of newcastle disease virus(NDV) F DNA vaccine with chicken IL-18 gene eukaryotic expression plasmid(pcDNA-chIL-18),F gene of NDV strain F48E9 was amplified by RT-PCR.The eukaryotic expression plasmid pcDNA-F(DNA vaccine) was constructed by inserting F gene into the pcDNA 3.1 vecter.11-day-old chickens were divided into 4 groups,10 chickens per group.14-day-old chickens were primarily immunized by injected intramuscularly and boosted immunized 14 days later after primary immunization,respectively with pcDNA-F,pcDNA-F combined pcDNA-chIL-18,ND attenuated vaccine and NS.On days 7,14,21 and 28 post-immunization,peripheral blood antibody response to NDV and T lymphocyte transformation ratio were detected by ELISA and LTT(MTT),respectively.Every chicken was challenged with 50LD50 NDV strain F48E9 at 29 day post immunization.The results showed that the pcDNA-F group obtained 3/10 protection,the pcDNA-F combined pcDNA-chIL-18 group obtained 5/10 protection.Their protection effect were less than that of ND conventional vaccine group(10/10).Antibody titers against NDV of the pcDNA-F combined pcDNA-chIL-18 group had little difference than that of the pcDNA-F group(P>0.05).T lymphocyte transformation ratio of the pcDNA-F combined pcDNA-chIL-18 group was significantly higher than that of the pcDNA-F group(P<0.05).It is concluded that pcDNA-F could produce some protection to the immunized chickens,and pcDNA-chIL18 could produce some immunoenhancing effects to the chickens immunized with pcDNA-F.更多还原