摘要
目的:观察过敏性哮喘患者单核细胞来源的树突状细胞接触变应原前后的表型及成熟状态的改变。方法:实验于2005-06/10在武汉大学医学院中心实验室进行。选择武汉大学人民医院门诊就诊的过敏性哮喘患者10例,以10名武汉大学医学院的健康志愿者为对照组。采用Ficoll-hypaque密度梯度离心法分离出两组受试者外周血单核细胞,加入重组人粒-巨噬细胞集落刺激因子(50μg/L)、重组人白细胞介素4(100μg/L),贴壁法诱导出树突细胞。各分成两份,一份不加屋尘螨变应原溶媒(刺激前),一份于培养的第6天加入屋尘螨变应原溶媒1mg/L(刺激后)。均于第7天收集细胞,用流式细胞仪检测刺激前后树突细胞表面CD11C,CD40,CD80,CD83,CD86的表达。结果:①接触变应原前:哮喘组树突状细胞表达CD80,CD86水平显著高于对照组[(2.4±0.5)%,(1.8±0.4)%;(14.8±4.0)%,(10.1±3.2)%,P<0.01],表达CD40水平显著低于对照组[(9.2±3.0)%,(13.1±3.8)%,P<0.01]。②接触变应原后:哮喘组树突状细胞表达CD80,CD86水平显著高于对照组[(6.2±2.1)%,(2.2±0.5)%;(19.2±4.8)%,(11.3±4.2)%,P<0.01],表达CD40水平显著低于对照组[(11.4±3.2)%,(16.2±4.1)%,P<0.05]。③哮喘组树突细胞接触变应原后CD80,CD86水平均高于接触前(P<0.01,0.05),CD11C,CD83表达差异无显著性。结论:哮喘患者单核细胞来源的树突状细胞表达CD80,CD86水平高,接触变应原后,表达CD80,CD86水平高于接触前和对照组,提示其表型向着Th2免疫和气道高反应性方向偏移,在哮喘发病机制中可能起促进作用。
AIM：To observe the changes of phenotypes and maturation of monoeytederived dendritic cell from allergic asthmatic patients before and after allergen contact.
METHODS： The experiment was conducted at the central laboratory of Medical College of Wuhan University between June and October 2005. Ten allergic asthmatic patients, who were diagnosed at the out-patient clinic of Renmin Hospital, Wuhan University, were selected. Ten healthy volunteers from Medical College of Wuhan University were regarded as control group. Monocytes of peripheral blood of the testees from the two groups were separated with Ficoll-hypaque density gradient centrifugation. Recombinant human histocytes colony stimulating factor （50 trg/L）, recombinant human interleukin 4 （100μg/L） were added. Dendritic cells were induced with adherence method. They were all assigned into two portion. Solvent of dermato-phagoides pterongssinus allergen was not added into one portion （before stimulation）. At the 6^th day after culture 1 mg/L solvent of dennatophagoides pterongssinus allergen was added （after stimulation）. Cells were collected at the 7^th day. Expressions of CDIIC,CD40,CD80,CD83 and CD86 at the surface of dendritic cell were detected with flow cytometer before and after stimulation.
RESULTS： ①Before contacting allergen： The levels of CD80 and CD86 in the asthma group were higher significantly than those in the control group [（2.4±0.5）%, （1.8±0.4）% ; （14.8±4.0）%, （10.1±3.2）% ,P 〈 0.01], and the level of CD40 was lower obviously than that in the control group [（9.2±3.0）% ,（13.1±3.8）%,P 〈 0.01]. ②After contacting allergen： The levels of CD80 and CD86 in the asthma group were higher significantly than those in the control group [（6.2±2.1）% , （2.2±0.5% ; （19.2±4.8）% , （11.3±4.2）%, P 〈 0.01）] , and the level of CD40 was lower obviously than that in the control group [（11.4±3.2）%,（16.2±4.1）% ,P 〈 0.05].③The levels of CD80 and CD86 after dendritic ceils contacting allergen in the asthma group were higher than those before contacting （P 〈 0.01, 0.05）. The expressions of CDI IC and CD83 had no significant difference.
CONCLUSION： The levels of CD80 and CD 86 of monocyte-derived dendritic cells in asthma patients are＇ high, After contacting allergen, the levels of CD80 and CD86 are higher than those before contacting allergen and those in the control group, It is indicated that its phenotype developed toward Th2 immunity and airway hyperreactivity polarization, which may play a stimulative role in asthmatic pathogenesis.
出处
《中国临床康复》
CSCD
北大核心
2006年第4期85-87,i0002,共4页
Chinese Journal of Clinical Rehabilitation
