鸡IL-18真核表达质粒与新城疫病毒F基因疫苗联合免疫探讨  

Immunization of Newcastle Disease Virus F Gene DNA Vaccine with Chicken IL-18 Gene Eukaryotic Expression Plasmid

作　　者：谢安新[1] 尹会方[1] 温纳相[1] 黄桂菊[1] 赵明秋[1] 郑海华[1] 黄青云[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《华南农业大学学报》2009年第2期82-85,89,共5页Journal of South China Agricultural University

基　　金：广东省自然科学基金(20043227)

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.To study the effect of immunization of newcastledisease virus （NDV） F DNA vaccine with chicken IL-18 gene eukaryotic expression plasmid （pcDNA-chlL-18）, F gene of NDV strain F4sE9 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-chlL-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 pcDNAcblL-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）. Tlymphocyte transformation ratio of the pcD-NA-F combined pcDNA-chlL-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.

关 键 词：新城疫病毒 F基因 基因疫苗 白细胞介素18基因 

分 类 号：S852.4[农业科学—基础兽医学]