重组HBHA蛋白对肺泡Ⅱ型上皮细胞细胞因子分泌的影响

Effect of recombinant heparin-binding hemagglutinin adhesion protein on secretion of cytokines in human alveolar type Ⅱ epithelial cells

目的研究重组HBHA蛋白对肺泡Ⅱ型上皮细胞分泌细胞因子的影响。方法原核表达并纯化重组HBHA,N端缺失HBHA(HBHA-ΔN)、C端缺失HBHA(HBHA-ΔC)三种蛋白,用以刺激A549细胞,并设置阴性对照,观察A549细胞分泌细胞因子的情况。结果 rHBHA蛋白可以显著提高A549细胞分泌IL-1β,IL-6,TNF-α,INF-γ的水平,而HBHA-△N和HB-HA-△C蛋白对A549细胞分泌IL-1β,IL-6,TNF-α,INF-γ和IL-10的作用无显著影响。结论 rHBHA能够促进IL-1β,IL-6,TNF-α,INF-γ分泌,N端和C端缺失后促进作用减弱。提示HBHA在诱导自然免疫以及抵抗结核菌感染中具有重要作用。

Objective To explore the influence of heparin-binding hemagglutinin adhesion（HBHA） protein on the seretion of cytokines in human alveolar type Ⅱ epithelial cells.Methods Recombinant HBHA（rHBHA）,HBHA-△C,HBHA-△N were expressed and extracted.A549 cells were treated with different concentrations of those three types of protein.The secretion of cytokines was detected using ELISA.Results IL-1β,IL-6,TNF-α,INF-γ were increased significantly in a dose-dependent manner after A549 cell were treated with rHBHA.While HBHA-△C and HBHA-ΔN had no significant effect on the secretion of IL-1β,IL-6,TNF-α,INF-γ of A549 cells.Levels of IL-10 did not change a lot in A549 cells after treated with rHBHA,HBHA-△C or HBHA-ΔN.Conclusion Recombinant HBHA could induce the secretion of IL-1β,IL-6,TNF-α,INF-γ in A549 cells,while the induction will be depressed after loss of N-terminal region and C-terminal region.The results suggest that HBHA serve as an antigen in induction of innate immune against tuberculosis.