不同剂量CpG-ODN佐剂与ALV-Jgp85重组蛋白联合免疫诱导种母鸡血清抗体和母源抗体的比较  

Comparison of Serum and Egg Yolk Antibodies Induced by Different Doses of CpG-ODN Adjuvant Combined with ALV-J gp85 Recombinant Proteins in Breeding Hens(Gallus domesticus)

作　　者：聂杰[1] 张丹丹[1] 井维芳[1] 任庆亚[1] 崔治中[1] 郭慧君[1] 李宏梅[1] 

机构地区：[1]山东农业大学动物科技学院,山东省生物工程与疫病防治重点实验室,泰安271018

出　　处：《农业生物技术学报》2014年第2期219-224,共6页Journal of Agricultural Biotechnology

基　　金：公益性行业(农业)科研专项经费资助(No.201203055)

摘　　要：为比较不同剂量非甲基化寡聚脱氧核苷酸(CpG-ODN)免疫佐剂与J亚群禽白血病病毒(Subgroup J Avian leukosis virus,ALV-J)gp85重组蛋白联合免疫后诱导母鸡血清抗体和母源抗体的效果,利用原核表达系统制备包含gp85基因的重组蛋白作为免疫原,与不同剂量CpG免疫佐剂联合接种成年海兰褐父母代种母鸡(Gallus domesticus),首免后2周加强免疫一次,于接种前和接种后每周动态检测血清抗体,接种后部分鸡只每周收集种蛋,检测卵黄抗体。结果显示,利用原核表达系统制备的gp85重组蛋白能与ALV-J单克隆抗体JE9特异性结合;与3个不同剂量CpG联合免疫成年种母鸡后均诱导鸡只产生一定效价的血清抗体,首免后28 d效价最高;所诱导的卵黄抗体效价首免后第4周最高。其中以50μg/羽剂量的CpG联合重组蛋白接种鸡只诱导的血清抗体和卵黄抗体效价和阳性比例最佳,100μg/羽剂量的CpG次之。3个不同剂量的CpG与ALV-J gp85重组蛋白联合免疫均能诱导母鸡产生一定的血清抗体和卵黄抗体,50μg/羽剂量的CpG联合免疫的效果明显优于较高剂量的免疫效果。本研究为使用CpG联合ALV-J gp85重组蛋白诱导母鸡产生母源抗体,阻断ALV-J早期感染提供科学依据。To compare the effects of three different doses of CpG oligonucleotide （CpG-ODN） adjuvants combined with subgroup J Avian leukosis virus （ALV-J） gp85 recombinant protein on serum and egg yolk antibodies in breeding hens（Gallus domesticus）, the recombinant protein containing ALV-J gp85 gene was prepared using the prokaryotic expression system and vaccinated combining with three different doses of CpG-ODN adjuvant into adult Hy-line Brown parent breeding hens, the booster immunization was conducted with two weeks interval. The serum antibodies and the egg yolk antibodies against ALV-J were detected weekly using ALV-J antibody kits from 1 day post inoculation to 49 day post inoculation and the data were statistically analyzed with ANOVA software. The results of protein gel electrophoresis showed that the recombinant protein containing gp85 gene of ALV-J was successfully prepared with prokaryotic expression system and the size of the recombinant protein was 45 kD; the Western blotting analysis indicated that the purified recombinant protein could specifically bind with the monoclonal antibody JE9 of ALV-J gp85 protein; the serum antibodies in breeding hens were successfully induced by three different doses of CpG combining with the recombinant gp85 protein, the serum antibody titers were increasing from the first post inoculation and reached a peak at the 28th day post first inoculation and then decreased slightly. Between three vaccinated groups, the antibody titers in the 50 μg CpG dose group were higher than those in the other two vaccinated groups. The ratio of positive antibodies in the 50 μg CpG dose group was up to 92%, but it was up to 80% in the 100 μg CpG dose group and only 67% in the 200 μg CpG dose group, respectively. The egg yolk antibodies against ALV-J in three vaccinated groups were successfully detected from the 3rd week to the 7th week post first inoculation; the antibody titers in the 50 μg CpG dose group were optimized at the 4th, the 6th and the 7th week post the first inoculat

关 键 词：非甲基化寡聚脱氧核苷酸(CpG-ODN)佐剂 J亚群禽白血病病毒(ALV-J)gp85重组蛋白 种母鸡 剂量 母源抗体 血清抗体 

分 类 号：S859.797[农业科学—临床兽医学]