串联表达FMDV多表位基因重组腺病毒免疫小鼠的效应研究

The Influence of Recombinant Adenovirus Expression Multiple Epitopes Genes of Foot and Mouth Disease Virus on Specific Immune Responses in Mice

目的检测口蹄疫重组腺病毒多表位疫苗(rAd5EGS)对小鼠特异性免疫的影响。方法 6周龄雌性BALB/c小鼠随机分为三组,每组15只;rAd5EGS和rAd5VP1以腹膜内接种的方式免疫小鼠,对照组小鼠注射PBS。通过MTT法检测小鼠T淋巴细胞增殖功能、ELISA法检测小鼠血清中IgG水平及IgG亚型(IgG1和IgG2a),定量ELISA法检测小鼠血清中IL-4和IFN-γ的表达水平。结果 rAd5EGS既能诱导小鼠T淋巴细胞增殖又能诱导小鼠特异性抗体的产生,其诱导小鼠淋巴细胞增殖能力明显高于rAd5VP1免疫组(P<0.05);IgG亚型和细胞因子检测结果显示,rAd5EGS免疫组小鼠血清中IgG2a的分泌水平高于rAd5VP1免疫组(P<0.05),rAd5EGS免疫组小鼠血清中IFN-γ的表达水平高于rAd5VP1免疫组(P<0.05)。结论rAd5EGS具有良好的免疫原性,且诱导Th1分化的能力高于rAd5VP1,为口蹄疫表位疫苗的研究奠定基础。

Objective To detect the influence of recombinant adenovirus rAd5EGS expressing FMDV multiple epitopes genes on specific immune responsed in mice. Methods 6 weeks female BALB/c mice were randomly divided into 3 groups： 15 mice per group, mice were inoculated intraperitoneal with recombinant virus rAdJEGS, and with rAd5VP1 and PBS as control. T lymphocyte proliferation function was tested by MTI＇. Levels of [gG,IgGt and IgG2a in the serum were detected by ELtSA. Expression levels of IL-4 and IFN -γ in the serum were quantitatively measured by ELISA. Results Recombinant adenovirus rAd5EGS not only induced T lymphocyte proliferation but also induced specific antibody secretion, which stimulated lymphocyte proliferation significantly higher than rAd5VP did（P 〈0.05 ）. The secretion level of IgG2a in the mice serum immunized with rAd5EGS was higher than those in rAd5VP1 immune group （P 〈 0.05 ） ;the level of expression of IFN - γ in the mice serum immunized with recombinant adenovirus rAd5EGS was higher than that inrAdSVP1 immune group （P 〈0.05） . Conclusion Recombinant adenovirus rAd5EGS had good immunogenicity,and it induced stronger Thl response than rAd5VP1 immune group did, which provided valuable sup- port for further development of FMD epitopes vaccine.