【摘要】 目的探讨树突状细胞(DCs)在曲霉菌免疫中的作用以及曲霉菌抗原冲击对DCs功能的影响。方法小鼠骨髓制备DCs,于小鼠尾静脉接种,以3H-TdR掺入法检测DCs刺激小鼠脾脏T细胞分化能力,ELISA方法检测IFN-γ和IL-12的浓度,电镜观察DCs的形态,同时进行DCs的表型测定。结果电镜下可见DCs细胞形态不规则,表面伸展出大量树突,与曲霉菌共同培养后胞内含有大量的烟曲霉孢子,部分孢子的膜被破坏;与烟曲霉孢子共培养24h后,DCs细胞表形CD40、CD80、CD86的表达明显增高,产生IL-12p70约(700.40±93.75)pg/ml,明显高于对照组(141.96±52.06)pg/ml;烟曲霉抗原冲击DCs回输小鼠的脾脏T细胞增殖能力明显增强,体外接受烟曲霉抗原24h产生IFN-γ(1084.33±238.04)pg/ml,明显高于单纯DCs接种小鼠的脾脏T细胞(345.98±32.75)pg/ml(p<0.01)。结论DCs能吞噬并破坏加热灭活的烟曲霉孢子,并趋于成熟,抗原呈递能力增加。更多还原

【Abstract】 Objective To assay the influence on the function of dendritic cells(DCs) pulsed with Aspergillus in the antiinfective immunity against Aspergillus.Methods DCs were generated from mice bone marrow cells and pulsed with the antigen of Aspergillus.Then DCs were transfered into mice via the lateral tail vein.Whether transfer of DCs pulsed with Aspergillus would lead to Aspergillus-specific proliferation of T cells in vivo was analyzed using ~ 3 H-TdR.The cytokine concentrations were assayed by a sandwich ELISA.With both light and scanning electron microscopy,DCs morphology was examined.Cell surface markers of DCs were analyzed by antibody staining and flow cytometry.Results A typical DCs morphology with characteristic veils,multiply elongated cytoplasmic processes and multivesicular organelles containing cell membrane-damaged bacteria were observed.DCs incubated with Aspergillus demonstrated an enhanced expression of the costimulatory molecules CD_ 40 ,CD_ 86 and CD_ 80 .Aspergillus-pulsed DCs secreted more IL-12 than that of non-pulsed.Spleen-derived T cells from mice immunized with Aspergillus-pulsed DCs showed increased proliferation compared to T cells from naive mice or mice that had received naive DCs.Spleen T cells from Aspergillus-pulsed DCs-immunized mice produced higher levels of IFN-γ upon in vitro restimulation with Aspergillus antigens,compared with T cells from naive spleen cells.Spontaneous production or an Ag-induced IFN-γ response was not detected in T cells derived from mice received i.v. heat-inactivated Aspergillus fumigatus(HAF) alone.Conclusions DCs can engulf and destroy HAF conidia.When pulsed with HAF,DCs changed their morphology and expression of the costimulatory molecules,enhanced their capacity to process antigen.DCs play major roles in host defense against Aspergillus infection.更多还原