IL-2真核质粒对乳酸杆菌载体口蹄疫VP1 DNA疫苗猪体免疫反应的分子佐剂效应  

Lactobacillus as a carrier in DNA vaccination against FMDV VP1 induce immune responses in swine which is enhanced by co-administration with swine IL-2

作　　者：李云岗[1] 田夫林[2] 周开锋[2] 高凤山[1] 李新生[1] 马慧玲[2] 夏春[1] 

机构地区：[1]中国农业大学动物医学院,北京100094 [2]山东省畜牧兽医总站,济南250022

出　　处：《中国农业大学学报》2006年第6期73-78,共6页Journal of China Agricultural University

摘　　要：为研究猪IL-2真核表达质粒对乳酸杆菌载体口蹄疫VP1 DNA疫苗(Lactobacillus acidophilusSFMD-1)猪体免疫的分子佐剂效应,通过RT-PCR方法从猪的肠系膜淋巴结克隆猪IL-2基因,基于pRc/CMV2载体,构建编码猪IL-2的真核表达质粒pCSIL2,以RT-PCR方法分析了该质粒转染PK15细胞以后的mRNA表达。结果显示预期的429 bp IL-2带仅出现在pCSIL2转染细胞。试验猪单独免疫L.acidophilusSFMD-1或者联合免疫pCSIL2质粒,后者能够增强FMDV-VP1特异的T细胞和B细胞反应,免疫后5周,单独免疫组和联合免疫组的抗体水平分别为1.50和1.66;免疫后6周,抗体水平分别上升到1.83和2.28;在T细胞增殖试验中,L.acidophilusSFMD-1单独免疫组和pCSIL2联合免疫组的刺激指数(SI)分别为2.00和2.70。结果表明,联合应用IL-2真核表达质粒是改进乳酸杆菌载体口蹄疫VP1 DNA疫苗免疫原性和效能的有效方法。In this paper, we studied the molecular adjuvant effect of IL-2 on DNA vaccine against FMDV VPt （Lactobacillus acidophilus SFMD-1） in swine, Swine IL-2 gene was cloned from mesentery lymph node by RT-PCR method, then it was ligated to eukaryotic expression plasmid pRc/CMV2 to produce pCSII_2, The expression of mRNA was analyzed in transfected PK15 cells by RT-PCR with specific primers. The predicted 429 bp band of IL-2 gene was appeared only in cells transfected with the pCSIL2, Swine were immunized with L. acidophilus SFMD-1 or co-administration with pCSIL2. Anti-FMDV-VPt-specific antibody titers were measured by the indirect ELISA method. Co-delivery with pCSIL2 plasmid was able to enhance both VPt-specific T cell and B cell response. Three weeks post primary immunization, swine vaccinated with L. acidophilus SFMD-1 alone and co-delivery with pCSIL2 groups reached 1,50 and 1.66, and then rose to 1.83 and 2.28 on three weeks of booster, respectively. In the MTT assay, the stimulation index （SI） of L. acidophilus SFMD-1 immunization alone and co-delivery group reached 2.00 and 2,70, respectively. Results clearly demonstrate that co-administration of IL-2 eukaryotic plasmid is a promising strategy for improving immunogenicity and efficacy of FMDV-VP1 DNA vaccine based on Lactobacillus as a carrierl.

关 键 词：DNA疫苗 口蹄疫 乳酸杆菌 质粒IL-2 基因 猪 分子佐剂 

分 类 号：S852.4[农业科学—基础兽医学]