【摘要】 研究重组复制缺陷腺病毒在小鼠肺内气道上皮细胞中的转基因表达水平及其介导小鼠γ 干扰素 (mIFN γ)的转基因表达对过敏原诱导的小鼠哮喘模型的预防性治疗作用。方法　带有LacZ表达基因的重组复制缺陷腺病毒 (AdCMVLacZ) ,经鼻腔吸入法和气管内注射法两种途径分别感染小鼠肺组织。C57小鼠经卵蛋白 (OVA)腹腔注射致敏后气道吸入激发建立哮喘模型 ,预防过敏反应组在激发前 48小时 ,麻醉下经鼻滴入 5× 1 0 9pfu含mIFN γcDNA的重组复制缺陷腺病毒 (AdCMVmIFN γ)。留取标本 ,监测被转基因的表达水平及其疗效。结果　AdCMVLacZ经鼻腔吸入法或气管内注射法均可感染小鼠气道和肺泡上皮细胞 ,并高效表达 β 半乳糖苷酶。经腺病毒载体介导的mIFN γ可以在体内、外高水平表达 (小鼠支气管肺泡灌洗液中mIFN (的含量在感染后4天达到 1 62 4 7± 1 3 2 1 5pg/ml) ,经预防性给予AdCMVmIFN γ可以明显降低小鼠哮喘模型中炎性细胞及嗜酸性粒细胞在肺组织中的浸润 ,支气管肺泡灌洗液中细胞总数由阳性对照组的 ( 2 1 6 6± 71 1 )× 1 0 3/ml,降至 ( 1 4 5±55 6)× 1 0 3/ml (P <0 0 5) ;嗜酸性粒细胞所占比例则由阳性对照组 75 1 3 %± 6 85% ,降至 9 0 0 %± 4 58% (P <0 0 0 1 )。结论　重组复制缺陷腺病毒可携更多还原

【Abstract】 To investigate adenoviral vector mediated exogenous gene expression in mouse lungs and the effect of mIFN γ transgene expression on allergen induced pulmonary eosinophil infiltration in a murine asthmatic model Methods LacZ marker gene was transduced into CD 1 mouse airway epithelial cells by installation of a replication deficient adenovirus with LacZ gene (AdCMVLacZ) 5×10 9 plaque forming unit (pfu) in the intratrachea or nostril C57 mice were sensitized intraperitoneally and challenged by aerosol with ovalbumin (OVA) to produce an asthmatic model AdCMVmIFNγ 5×10 9 pfu was administered via nostril in asthmatic mice 48 h before OVA challenge Sera, bronchial alveolar lavage (BAL) and lungs were recovered 48 h after OVA challenge Results After administration with AdCMVLacZ by intratracheal installation or nose drop, the lungs revealed a high level of widespread LacZ transduction with X gal staining, mainly along airways IFN γ via adenoviral vector transduction could be overexpressed both in vitro and in vivo (1624 7±1321 5 pg/ml in BAL 96 h after AdCMVIFNγ infection) In AdCMVIFNγ treated asthmatic models, histological evaluation revealed marked suppression of eosinophil peribronchial and perivascular infiltration; the recoverable percentage of eosinophils in BAL was an average of 9 00%±4 58%, which was a statistically significant decrease versus that of the positive control group (75 13%±6 85%) ( P <0 001) The total cell number in BAL ((145±55 6)×10 3 cells/ml) in AdCMVmIFNγ treated mice also was tremendously reduced compared to the positive control group ((216 6±71 1)×10 3 cells/ml) Conclusions Adenoviral vector was able to overexpress exogenous gene in murine lungs IFN γ overexpression via adenoviral vector in pulmonary epithelia in vivo can abrogate allergen induced eosinophilic infiltration in lungs in an asthmatic model, which may suggest a new preventively therapeutic method for cytokine immunogenetic transfer in allergic asthma更多还原