| 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。 |
| 关键词: DNA疫苗,口蹄疫,乳酸杆菌,质粒IL-2,基因,猪,分子佐剂 |
| DOI：10.11841/j.issn.1007-4333.2006.06.155 |
| 投稿时间：2006-05-25 |
| 基金项目: |
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| 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 |
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| Abstract: |
| In this paper,we studied the molecular adjuvant effect of IL-2 on DNA vaccine against FMDV VP1(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 pCSIL2.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-VP1-specific antibody titers were measured by the indirect ELISA method.Co-delivery with pCSIL2 plasmid was able to enhance both VP1-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. |
| Key words: foot-and-mouth disease,DNA immunization,Lactobacillus,plasmid IL-2,gene,swine,molecular adjuvant |
