论文部分内容阅读
目的:检验b型流感嗜血杆菌结合疫苗在体外能否刺激人气道上皮细胞hBD-2表达,并探讨人工诱导防御素表达的方法。方法:体外分离、培养原代人气道上皮细胞。b型流感嗜血杆菌疫苗刺激原代人气道上皮细胞后,用RT-PCR和ELISA法检测培养上清液中hBD-2蛋白的表达。收集刺激后的细胞培养上清液进行抑菌实验。结果:浓度>1μg/ml的b型流感嗜血杆菌结合疫苗刺激原代人气道上皮细胞12 h后,可诱导hBD-2 mRNA的表达,与对照组相比较有显著差异(P<0.01),无剂量依赖性,同时细胞培养上清液中hBD-2蛋白的表达上调。1μg/ml b型流感嗜血杆菌结合疫苗浓缩后的细胞培养上清液具有抑菌作用。结论:一定浓度(1μg/mL)以上的b型流感嗜血杆菌结合疫苗作用于人原代气道上皮细胞可诱导hBD-2的表达,为人工方法刺激机体产生人β防御素-2,对抗病原菌感染提供新的策略。
Objective: To test whether Haemophilus influenzae type b conjugate vaccine can stimulate the expression of hBD-2 in human airway epithelial cells in vitro and to explore the method of artificially induced defensin expression. Methods: Primary human airway epithelial cells were isolated and cultured in vitro. After the Haemophilus influenzae type b vaccine stimulated the primary human airway epithelial cells, the expression of hBD-2 protein in the culture supernatant was detected by RT-PCR and ELISA. The stimulated cell culture supernatants were collected for antibacterial experiments. Results: The hBD-2 mRNA expression in primary human airway epithelial cells stimulated with the Haemophilus influenzae type b conjugate vaccine at a concentration of> 1μg / ml for 12 hours stimulated the expression of hBD-2 mRNA significantly compared with the control group (P <0.01) No dose-dependent increase in hBD-2 protein expression in cell culture supernatants. 1μg / ml Haemophilus influenzae type b conjugate vaccine enriched cell culture supernatant has antibacterial activity. CONCLUSION: The hBD-2 expression induced by the Haemophilus influenzae type b conjugate vaccine above a certain concentration (1μg / mL) can induce the expression of hBD-2 in human primary airway epithelial cells and stimulate the body to produce human β-defensin-2 Pathogen infection provides new strategies.