摘要
目的分析贝氏柯克斯体(Cb)的Coml和热休克蛋白B(HspB)对体外诱导培养的人源树突状细胞(DC)表面分子和细胞因子表达的影响。方法分别以20μg/mL的重组蛋白Com1和HspB、6μg/mL的大肠埃希菌LPS为终浓度刺激培养5d的DC,24h后流式细胞仪检测DC表面成熟标志分子CD83和T淋巴细胞CD58、CD54、CD40、CD80、CD86以及细胞因子IL-10和IL-12的表达水平。多重比较应用SNK检验。结果Com1刺激能有效促进体外诱导培养的DC成熟,DC细胞表面成熟标志分子CD83和T淋巴细胞活化辅助分子CD54、CD58、CD80、CD86和CD40等处于高表达水平,表达水平均〉80%,其中CD83表达水平与大肠埃希菌LPS刺激DC相似,CD54和CD86的表达量略高于大肠埃希菌LPS刺激DC,其他分子的表达量显著高于大肠埃希菌LPS刺激DC(均P〈0.05)。Com1刺激后,细胞内IL-12水平从无增至9%左右。HspB刺激不能促进DC表型成熟,HspB刺激的DC胞内IL-10水平达6%左右。Com1和HspB的先后刺激,使IL-12水平几乎为0,IL-10水平降至2%以下。结论Com1为DC有效的促成熟抗原,被Com1激活的DC具备启动T淋巴细胞免疫的功能条件。
Objective To investigate the change of surface molecules and cytokine expressions of dendritic cells (DC) stimulated with recombinant proteins Com1 and heat shock protein B (HspB) of Coxiella burnetii (Cb). Methods The DC cultured for 5 d were stimulated with 20 μg/mL recombinant protein Com1, 20 μg/mL HspB or 6 μg/mL E. coli lipopolysaccharide (LPS). Twenty- four hours later, the surface mature marker, CD83, and activation-associated markers of T lymphocytes, CD58, CD54, CD40, CD80 and CD86, and expression levels of interleukin (IL)-10 and IL-12 of DC were detected by fluorescence activated cell sorter (FACS). Multiple comparisons were performed by using Student-Newman-Keuls test. Results Coral could induce DC maturation efficiently in vitro. Human monocyte-derived DC exhibited significantly higher expression levels of surface molecules including CD83, CD54, CD58, CD80, CD86, and CD40, which were all 〉80%. CD83 expression induced by Coral was similar with that induced by E. coli LPS, while CD54 and CD86 expressions were slightly higher than those induced by E. coli LPS, and expressions of other molecules were significantly higher than those induced by E. coli LPS (all P〈0. 05). After Coral stimulation, intracellular IL-12 level increased to 9 % from zero. HspB could not induce DC maturation in vitro. The intracellular IL-10 level was 6% after HspB stimulation. DC pulsed with Com1 and HspB exhibited intracellular IL-12 level of zero and IL-10 level of 〈2%. Conclusion Com1 but not HspB can efficiently activate DC and Com1-activated DC can drive T cells toward Th1 cell development due to a high level of IL-12 production.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2011年第11期653-658,共6页
Chinese Journal of Infectious Diseases