联合肌注猪IL2真核质粒对PPV VP2-PCV2 ORF2核酸疫苗免疫效果的影响  

Effects of combined intramuscular injection of the swine IL2 eukaryon plasmid on the immune responses of PPV VP2-PCV2 ORF2 DNA vaccine

作　　者：朱玲[1] 郭万柱[1] 徐凯[1] 王印[1] 陈燕凌[1] 唐玉香[1] 

机构地区：[1]四川农业大学动物生物技术中心动物疫病与人类健康实验室,四川雅安625014

出　　处：《中国兽医学报》2011年第2期145-150,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30500019);“长江学者和创新团队发展计划”创新团队项目(IRT0848)

摘　　要：将猪IL2基因和PPV VP2基因、PCV2 ORF2截短基因克隆至真核表达载体pCI-neo,构建了重组质粒pCI-ORF2-VP2和pCI-IL2,用脂质体介导法将pCI-ORF2-VP2质粒转染PK15细胞,采用间接免疫荧光法检测在体外的表达情况。并以小鼠为动物模型,将pCI-IL2和pCI-ORF2-VP2质粒联合肌注免疫小鼠,相同剂量免疫2次,间隔2周,同时设立pCI-ORF2-VP2、pCI空载体、猪细小灭活苗、圆环亚单位苗对照,检测免疫小鼠脾脏淋巴细胞的转化功能,外周血T淋巴细胞亚群的动态变化及血清抗体的滴度。结果显示,pCI-ORF2-VP2在PK15细胞中能正常表达,联合免疫组在第28天能够检测到PPV和PCV2抗体,第21~42天诱导淋巴细胞转化和CD4＋、CD8＋细胞的数量高于或显著高于pCI-ORF2-VP2质粒组。结果表明,猪IL2质粒联合肌注可以提高VP2/ORF2核酸疫苗的免疫效果。The swine IL2 gene、PPV VP2 gene and the PCV2 ORF2 truncated gene（47-207 aa） were cloned into the expression vector pCI-neo respectively,then we constructed the expression plasmid pCI-ORF2-VP2 and pCI-IL2.After transfected PK15 cells by lipofectamine,the ORF2/VP2 expressed product was detected by immunofluore assay.To study the immune effects of DNA vaccine in vitro and in vivo,mice were used as the animal model.The combined plasmid of pCI-ORF2-VP2 and pCI-IL2 was immunized by intramuscular injection,two times in the same dose and two weeks interval.In the mean time,the pCI-ORF2-VP2 plasmid、 PPV vaccine、PCV vaccine and empty vector were set up as the control groups.Anti-PPV/PCV2 antibodies were measured by ELISA,lymphocyte proliferation activity was detected using MTT method and the dynamic monitoring of the T-lymphocyte subsets respectively.The results showed that the fusion protein can express normally and the anti-PPV/PCV2 antibodies were detected after 4 weeks.The reaction of the splenic T lymphocytes to the ConA stimulation and the immune function of inducing the CD4＋,CD8＋ T lymphocyte of the combined group were higher or significantly higher than those of the pCI-ORF2-VP2 group after three weeks.The result indicated that the PPV VP2-PCV2 ORF2 vaccine could induce higher response of immune response and IL2 eukaryon expression plasmid was effective adjuvant which could significantly enhances the immunogenicity of VP2/ ORF2 DNA vaccine.

关 键 词：细小病毒 圆环病毒2型 VP2基因 ORF2基因 IL2基因 

分 类 号：S852.65[农业科学—基础兽医学]