目的研究流行性感冒病毒裂解疫苗刺激人原代气道上皮细胞后能否促进人β防御素2(human β—defensin-2,HBD-2)的表达及分泌,探讨人工诱导增强防御素表达的方法。方法分离培养人原代气道上皮细胞,以不同浓度流行性感冒病毒裂解疫苗刺激人原代气道上皮细胞,分别于刺激后6、12、24h提取细胞总RNA及培养细胞上清液,RT—PCR、ELISA检测hBD-2的mRNA和蛋白表达,纸片法观察杀菌效果。结果不同浓度流行性感冒病毒裂解疫苗刺激气道上皮细胞6h后,各组细胞中hBD-2的mRNA和蛋白表达与对照组相比无明显变化;12h后,疫苗浓度〉1.0μg/ml的各组细胞中mRNA表达和蛋白水平显著升高,差异有统计学意义(P〈0.05),但24h后均下降。1.0μg/ml流行性感冒病毒裂解疫苗作用12h后的细胞培养上清液有明显的杀菌效果。结论一定浓度的流行性感冒病毒裂解疫苗刺激人原代气道上皮细胞,可促进hBD-2 mRNA的表达和hBD-2蛋白浓度的增加,为人工方法刺激机体产生hBD-2,对抗病原菌感染提供新的方向。
Objective To investigate the expression of hBD-2 in human bronchial epithelial cells stimulated by influenza virus split vaccine. Methods Human bronchial epithelial cells were isolated, cultured and then stimulated by influenza virus split vaccine at various concentrations. The hBD-2 mRNA and protein in cultured cells were detected by RT-PCR and ELISA methods respectively. The bacteriostasis effects were tested with supernatant of cell culture after vaccine stimulation. Results After stimulation with influenza virus split vaccine 〉 1.0 μg/ml for 12 h the expression of hBD-2 mRNA and protein increased compared to controls ( P 〈 0.05 ) ; the expression was decreased at 24 h. The condensed supernatant acquired the ability of bacteriostasis. Conclusion Influenza virus split vaccine can significantly enhance the expression of hBD-2 in human bronchial epithelial cells and may have a role in bacteriostasis.