山地乌骨鸡IL-2、IL-15、IL-18及IFN-γ真核表达载体构建及其对IBV DNA疫苗免疫增强作用研究  

Construction of eukaryotic expression plamids encoding chicken IL-2, IL-15, IL-18 and IFN-γ and study on their immunoenhancement on avian infectious bronchitis DNA vaccine

作　　者：胡慧琼[1] 王红宁[1,2] 周生[1] 李晓英[1] 黄勇[1] 柳萍[1] 徐永莉[1] 

机构地区：[1]四川农业大学动科院动物预防医学生物工程实验室,雅安625014 [2]四川大学“985工程”西南资源环境与灾害防治科技创新平台,四川大学生命科学学院,成都610064

出　　处：《高技术通讯》2006年第10期1056-1062,共7页Chinese High Technology Letters

基　　金：863计划（2003AA241120）资助项目

摘　　要：利用RT-PCR法从刺激的山地乌骨鸡的外周淋巴细胞及脾脏组织总RNA中扩增出与鸡IL-2、IL-15、IL-18及IFN-γ基因预期大小的DNA片段,并与pcDNA3.1连接构建了真核表达质粒pDNAIL2、pDNAIL15、pDNAIL18及pDNAIFN-γ.研制了4种分子免疫佐剂,对通过检测血清总IgG值、CD4＋、CD8＋百分含量,研究了４种分子免疫佐剂对禽传染性支气管炎DNA疫苗的免疫增强作用.并对pDNAIL2在鸡胚成纤维细胞中的表达进行了研究.研究发现,所有分子佐剂和DNA疫苗联合接种试验组鸡血清总IgG值、CD4＋T及CD8＋T淋巴细胞含量从5天后就一直高于DNA疫苗单独注射组,并在15天和21天时出现差异显著（P＜0.05）或极显著（P＜0.01）.结果表明,IL-2、IL-15、IL-18及IFN-γ对IBV DNA疫苗有免疫增强作用,且pDNAIL2在体外得到了表达.The specific DNA fragment of chicken interlerkin-2 （CHIL-2）, interleukin- 15 （ CHIL- 15 ）, interleukin-18 （ CHIL- 18） and intefferon-γ（CHIFN-γ）were amplified from peripheral blood lymphoeytes and spleen stimulated with ConA. The eukaryotie expression plasmids of Chicken IL-2, CHIL-15, CHIL-18 and CHIFN-γ were constructed by subeloning CHIL-2, CHIL-15, CHIL-18 and CHIFN-γ into peDNA3.1（ ＋ ） .They were pDNAIL2, pDNAIL15, pDNAIL18 and pDNAIFN-γ. The immunoenhaneement effect of pDNAIL2, pDNAIL15, pDNAIL18 and pDNAIFN-γ on DNA vaccine of Avian Infectious Bronchitis were observed by the test of total IgG level of semrn, percentage of CD4/CD8 T lymphoeytes from peripheral blood, and the in vitro expression of pDNAIL2 was studied. The result indicated that pDNAIL2, pDNAIL15, pDNAIL18 and pDNAIFN-γ have the immunoenhaneement effect on IBV DNA vaccine. It can not only enhance the cellmediated immune response obviously,but also enhance the humoral response, thus improve the protective rate of IBV DNA vaccine, and pDNAIL2 can be expressed in vitro.

关 键 词：山地乌骨鸡 白细胞介素2 白细胞介素15 白细胞介素18 γ-干扰素 真核表达载体 DNA疫苗 

分 类 号：S858.31[农业科学—临床兽医学]