过敏性哮喘患者树突状细胞的趋化因子受体及表型研究

目的:探讨过敏性哮喘患者和健康人外周血单核细胞来源树突状细胞(DC)负荷抗原后表达组成性趋化因子受体(CCR7)和MHCⅡ类分子(HLA-DR)的差异,为进一步研究过敏性哮喘的发病机制及治疗提供新的思路。方法:分别从过敏性哮喘患者(哮喘组)和健康人(对照组)外周血分离单个核细胞并获取贴壁细胞,经重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和重组人白细胞介素-4(rhIL-4)体外培养为未成熟DC,加入Der p 1使其负荷抗原;观察培养的DC的形态变化,并用流式细胞仪检测两组负荷抗原的DC表面趋化因子受体(CCR7)及MHCⅡ类分子(HLA-DR)的表达。结果:与对照组相比,哮喘组负荷抗原的DC表达较高水平CCR7(P<0.01)及HLA-DR(P<0.01);两组CCR7和HLA-DR均呈正相关(哮喘组r=0.73,P<0.05;对照组r=0.76,P<0.05)。结论:由于过敏性哮喘患者负荷抗原的DC与健康人相比在趋化因子受体及成熟标志物表达方面存在明显差异,从而趋化因子反应性及成熟情况存在差异,后者成为哮喘发生的重要机制之一。

Study of Chemokine Receptors and Phenotypes on Monocyte-derived Dendritic Cells from Allergic Asthmatic Patients

Objective: To explore the difference in expression of constitutive chemokine receptors and MHCⅡ molecules expressed by Der p 1-pulsed monocyte-derived dendritic cells(DCs) generated from both healthy donors and asthmatic patients sensitive to Dermatophagoides pteronyssinus(Dpt),and to establish experimental foundation for further research of pathogenesis and therapeutics in allergic asthma.Methods: Mononuclear cells were isolated from peripheral blood of healthy donors and asthmatic patients,and the adherent cells were harvested and cultured with the combination of rhGM-CSF and rhIL-4 as immature DCs(iDCs),then the generated cells were incubated for 24 hours with 1 mg/L Der p 1(allergen of Dermatophagoides pteronyssinus).The expression of CCR7 and HLA-DR on Der p 1-pulsed DCs was analyzed by flow cytometry(FCM).Results: Compared with control group,Der p 1-pulsed DCs from asthmatic group had increased the expression of CCR7(P<0.01) and HLA-DR(P<0.01).There was positive correlation between CCR7 and HLA-DR in both groups(both P<0.05).Conclusion: Because of increased expression of CCR7 and HLA-DR on Der p 1-pulsed DCs from asthmatic patients,there is a difference in chemokine responsiveness and maturation of DCs between asthmatic patients and healthy donors,which is one of important pathogenesis for allergic asthma.