目的 研究青春型双歧杆菌对过敏性哮喘儿童外周血单个核细胞(PBMC)来源的树突状细胞(DC)表达CD86和HLA-DR及其分泌IL-1β、IL-6、IL-10、IL-12、IL-23和IFN-γ的影响.方法 从15名过敏性哮喘儿童和15名非哮喘儿童的外周血单个核细胞诱导生成未成熟DC,分别加入青春型双歧杆菌或脂多糖(LPS)后继续培养DC 2天,①每天于倒置显微镜下观察DC形态;②用流式细胞仪检测各组DC表面CD86和HLA-DR的分子表达;③用ELISA方法检测培养上清中IL-1β、IL-6、IL-10、IL-12、IL-23和IFN-γ的水平.结果 ①DC经双歧杆菌和LPS分组处理后第2天,于倒置显微镜下观察到双歧杆菌组和LPS组的DC突起均较空白组明显,具有突起的细胞数量较空白组增多,而双歧杆菌组与LPS组比较,LPS组有突起的细胞数量明显增多,提示DC过度生长.②双歧杆菌刺激后,哮喘儿童DC表面CD86表达明显增高,HLA-DR表达无明显变化,非哮喘儿童CD86和HLA-DR表达均无明显变化;LPS刺激可明显增加哮喘儿童和非哮喘儿童CD86和HLA-DR的表达.③双歧杆菌刺激哮喘儿童DC分泌IL-12、IFN-γ、IL-1β及IL-6水平增高,刺激非哮喘儿童DC分泌IL-12、IL-10、IL-1β及IL-23水平增高.结论 ①过敏性哮喘儿童DC表面CD86的表达可能存在缺陷,双歧杆菌能适度上调其表达,在DC的成熟过程中可能起调节作用.②双歧杆菌能刺激过敏性哮喘儿童DC分泌IL-12和IFN-γ,从而改变Th2优势分化,纠正Th1/Th2失衡.③过敏性哮喘儿童DC分泌IL-10可能存在缺陷,从而影响免疫耐受的形成.④双歧杆菌可能通过刺激哮喘儿童DC分泌IL-1β及IL-6增高,达到促进Th17细胞分化的作用.%Objective ①To study the effects of bifidobacterium on the expression of CD 86 and HLA-DR、 secretion of IL-1β、IL- 6、IL-10、 IL-12、IL-23 and IFN-γin dendritic cells (DCs) derived from peripheral blood mononuclear cells (PBMC ) of the children with allergic asthma.②investigate the effect of bifidobacterium on in dendritic cells (DC) derived from the allergic asthma children peripheral blood monocytes(PBM C ) .Methods The immature DCs were induced from the PBMC derived from 15 allergic asthma and 15 non-asthma children,then the immature DCs were cocultured with the bifidobacterium or LPS respectively .①Observe the shape of DC with the microscope everyday;②The expression of CD 86 and HLA-DR of DCs were measured by flow cytometer ;③ IL-1β、 IL-6、 IL-10、IL-12、IL-23 and IFN-γ secreted by DC were measured by ELISA .Results ①The second day ,The ecphym as of DCs control by LPS or Bifidobacterium is obviously more than blank observed with the microscope ,And the quantity of cells with ecphym as is more than the blank too ,However,The quantity of cells with ecphym as control by LPS is obviously more than that control by Bifidobacterium , From the above-mentioned ,we can judge that the DCs control by LPS is overgrowth.②Atter the pretreatment with Bifidobacterium adolescent ,the expression of CD 86 of DCs from the allergic patients was sigificantly increased ,but the expression of HLA-DR was not changed .The pretreatment with Bifidobacterium adolescent have no effects on the expression of CD 86 and HLA-DR on the nonasthm a children .The pretreatment with LPS may increase the expression of CD 86 and HLA-DR from both the allergic patients and non-asthma children signigicantly .③The pretreatment with the Bifidobacterium adolescent may increase significantly the level of IL- 12、 IFN-γ、IL-1β and IL-6 in the patients with allergic asthma,and the level of IL-12、IL-10、 IL-1βand IL-23 in the non-asthma children .Conclusion ①Bifidobacterium adolescent could stimulate the mature of DCs .The expression of CD 86 of DCs from the patients with allergic asthma may be in paired ,the Bifidobacterium adolescent may up-regulate the expression of CD 86 appropriately .②The Bifidobacterium adolescent could stimulate the DCs from the patients with allergicasthm a to secret the IL-12 and IFN-γ ,then may alter Th2 dominant differentiation and retrieve the Th1/Th2 in balance .③The secretion of IL-10 of DCs from the patients with allergic asth- mamay be defect,Then may influence the form of immune to lerance .④The Bifidobacterium adolescent may also improve the Th17 cell differentiation by stimulated DC to increase the level of IL-1βand IL-6.
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