鸡IL-18真核表达质粒及原核表达蛋白对IBD死苗的免疫增强作用
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摘要
将80只14日龄SPF鸡随机分为4组,Ⅰ组为对照组,Ⅱ组为单纯疫苗试验组,Ⅲ组为质粒试验组,Ⅳ组为蛋白试验组。通过T淋巴细胞增殖试验、ELISA抗体检测和攻毒保护试验研究其对IBD灭活疫苗的免疫增强作用。结果显示,从一免后21 d起,Ⅲ组和Ⅳ组都与Ⅱ组T淋巴细胞增殖和抗体水平差异显著(P<0.05),且Ⅳ组效果最好。一免42 d后进行攻毒保护性试验,Ⅱ组、Ⅲ组和Ⅳ组保护率依次为60%、80%和75%。结果表明,鸡IL-18的真核表达质粒和原核表达蛋白都具有显著增强IBD灭活疫苗免疫效果的作用。
A total of 80 SPF chickens were randomly divided into four groups. The group Ⅱ,Ⅲ and Ⅳ were immuned with only vaccine,vaccine and plasmid and vaccine and protein,respectively. For the control group,normal saline was used. The immunologic effects were detected by T lymphocyte proliferative response,ELISA antibody detection and protective efficacy expreriment. It presented significant difference( P < 0. 05) among group Ⅲ,group Ⅳ and group Ⅱafter 21 days post-immunization in T lympholeukocyte proliferative response and antibody level detection of group Ⅳ is the best. The protection of group Ⅱ,group Ⅲ and Ⅳ was 60 %,80 % and 75 %. The results indicated that chicken IL-18 eukaryotic expression plasmid and Prokaryotic protein had the ability to obviously enhance the immunologic effect of IBD inactivated vaccine.
A total of 80 SPF chickens were randomly divided into four groups. The group Ⅱ,Ⅲ and Ⅳ were immuned with only vaccine,vaccine and plasmid and vaccine and protein,respectively. For the control group,normal saline was used. The immunologic effects were detected by T lymphocyte proliferative response,ELISA antibody detection and protective efficacy expreriment. It presented significant difference( P < 0. 05) among group Ⅲ,group Ⅳ and group Ⅱafter 21 days post-immunization in T lympholeukocyte proliferative response and antibody level detection of group Ⅳ is the best. The protection of group Ⅱ,group Ⅲ and Ⅳ was 60 %,80 % and 75 %. The results indicated that chicken IL-18 eukaryotic expression plasmid and Prokaryotic protein had the ability to obviously enhance the immunologic effect of IBD inactivated vaccine.
引文
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[11]李宏梅,胡敬东,马凤龙,等.鸡白细胞介素-18(Chl L-18)重组蛋白的生物学活性检测[J].农业生物技术学报,2007,15(1):5-10.
[12]王新华,孔娜,胡敬东,等.鸡IL-18原核表达蛋白及真核表达质粒对IBD灭活疫苗免疫增强作用的研究[A].中国畜牧兽医学会动物传染病学分会教学专业委员会第六届第12次学术研讨会论文集[C].2010:148-152.
[2]胡敬东,崔治中,范伟兴,等.鸡白细胞介素18成熟蛋白全长基因的克隆与序列测定[J].中国兽医学报,2004,24(2):119-121.
[3]胡敬东,崔治中,赵宏坤.鸡IL-18c DNA的克隆及其在大肠杆菌中的高效表达[J].畜牧兽医学报,2005,36(3):264-268.
[4]Muller H,Islam M R,Raue R.Research on infectious bursal disease-the past,the present and the future[J].Vet Microbiol.2003,97(1-2):153-165.
[5]Dolz R,Majo N,Ordonez G,et al.Viral genotyping of infectious bursal disease viruses isolated from the 2002 acute outbreak in Spain and comparison with previous isolates[J].Avian Dis.,2005,49(3):332-339.
[6]Van den Berg T.P.Acute infectious bursal disease in poultry:a review[J].Avian Pathol.,2000,29(3):175-194.
[7]Winfried G J,Hanneke I V,Nicolette C S,et al.Potentiation of humoral immune responses to vaccine antigens by recombinant chicken IL-18(r Ch IL-18)[J].Vaccine.2005,23:4212-4218.
[8]Iinuma H,Okinaga K,Fukushima R.Superior protective and therapeutic effects of IL-12 and IL-18 gene-transduced dendritic neuroblastoma fusion cells on liver metastasis of murine neuroblastoma[J].J Immunol.2006,176(6):3461-3469.
[9]郝牧,鲍朗,高蕾.人IL-12与结核分枝杆菌抗原ESAT-6联合基因疫苗的免疫效果观察[J].微生物学报,2007,47(3):477-481.
[10]曹素芳,黄青云,韩先干,等.禽巴氏杆菌C48-1外膜蛋白H基因与鸡IL-18基因真核共表达载体的构建与表达[J].中国预防兽医学报,2006,26(4):382-384.
[11]李宏梅,胡敬东,马凤龙,等.鸡白细胞介素-18(Chl L-18)重组蛋白的生物学活性检测[J].农业生物技术学报,2007,15(1):5-10.
[12]王新华,孔娜,胡敬东,等.鸡IL-18原核表达蛋白及真核表达质粒对IBD灭活疫苗免疫增强作用的研究[A].中国畜牧兽医学会动物传染病学分会教学专业委员会第六届第12次学术研讨会论文集[C].2010:148-152.