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目的 探讨尘螨主要变应原DNA疫苗对尘螨提取液诱导的小鼠肺部变应性炎症的免疫治疗作用。方法 分别构建表达Derp1和Derp 2的真核表达质粒pCMV Derp 1和pDerp 2 IRES Derp1。 30只BALB/c小鼠随机分为正常组 (6只 )、对照组 (12只 )、单质粒组 (6只 )和混合组 (6只 )。后三组分别用空白质粒、pDerp 2 IRES Derp 1或混合的pDerp 2 IRES Derp 1和pCMV Derp 1免疫 ,4周后用尘螨提取液致敏、激发 ,观察肺部炎症、计分、肺泡灌洗液 (BALF)细胞总数和分类计数 ;酶联免疫吸附法 (ELISA)检测BALF中白细胞介素 4 (IL 4 )和γ干扰素 (IFN γ)水平。结果 细胞总数和嗜酸细胞(EOS)计数 :混合组 [(6± 4 )× 10 5/ml、0 0 5± 0 0 4 ]和对照组 [(2 1± 13)× 10 5/L、1 80± 1 39]比较差异有显著性 (P <0 0 5 ) ;IL 4水平 :混合组 [(16 8± 2 33)g/L]与对照组 [(5 38± 2 5 6 )g/L]比较差异有显著性 (P<0 0 5 ) ,各组间IFN γ比较差异无显著性 (P >0 0 5 )。结论 尘螨主要变应原的DNA疫苗可有效抑制尘螨提取液诱导的肺部变应性炎症 ,能表达两个主要变应原的DNA疫苗其抑制变应性炎症效果优于只表达一种变应原的疫苗。
Objective To investigate the immunotherapy effect of dust allergen DNA vaccine on allergic inflammation in mice induced by dust mite extract. Methods The eukaryotic expression plasmids pCMV Derp 1 and pDerp 2 IRES Derp1 expressing Derp1 and Derp 2 were constructed respectively. Thirty BALB / c mice were randomly divided into normal group (n = 6), control group (n = 12), single plasmid group (n = 6) and mixed group (n = 6). The latter three groups were respectively immunized with blank plasmid, pDerp 2 IRES Derp 1 or mixed pDerp 2 IRES Derp 1 and pCMV Derp 1, and sensitized with dust mite extract four weeks later. The lung inflammation, scoring, alveolar perfusion The total number of BALF cells and their counts were counted. The levels of interleukin 4 (IL 4) and interferon gamma (IFN γ) in BALF were detected by enzyme linked immunosorbent assay (ELISA). Results The number of total cells and eosinophils (EOS) counts in the mixed group [(6 ± 4) × 10 5 / ml and 0 05 ± 0 0 4] and the control group [(2 1 ± 13) × 10 5 / 1 80 ± 1 39] (P <0 05). IL 4 levels were significantly higher in the mixed group (16 8 ± 2 33 g / L) than those in the control group [(5 38 ± 2 56) g / L] (P <0 05). There was no significant difference in IFN γ between groups (P> 0.05). Conclusion The DNA vaccine of the main allergen of dust mite can effectively inhibit the lung allergic inflammation induced by dust mite extract. The DNA vaccine that can express two major allergens is better than the one that only expresses allergic inflammation Allergen vaccine.