Functional expression of granzyme B in human plasmacytoid dendritic cells: a role in allergic inflammation
Functional expression of granzyme B in human plasmacytoid dendritic cells: a role in allergic inflammation
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DOI：
10.1111/j.1365-2222.2010.03499.x
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发表时间：
2010-07-01
影响因子：
6.1
通讯作者：
Virchow, J. C.
中科院分区：
文献类型：
--
作者：
Bratke, K.;Nielsen, J.;Virchow, J. C.
背景浆细胞样树突状细胞(PDCs)参与多种免疫功能。本研究的目的是系统分析细胞毒颗粒蛋白在人肺树突状细胞中的表达和调控。方法采用RT-PCR、流式细胞仪和荧光显微镜等方法分析细胞毒蛋白在人肺树突状细胞的表达。以K562细胞为靶细胞的流式细胞仪细胞毒实验证实了这些蛋白的功能性表达。为了分析PDC来源的细胞毒蛋白在传染病和变态反应性疾病中的调节作用，在Toll样受体(TLR)7/9配体刺激后和节段性变应原激发的人哮喘模型中分析了PDC。结果PDC特异性表达颗粒酶B(GRB)，而不表达颗粒酶A、H、K、M和穿孔素，IL-3刺激可上调这种GRB的表达。此外，IL-3刺激的pDC以GRB和caspase依赖的方式杀伤K562细胞。TLR7/9配体可显著抑制pDC中GRB的表达。相反，在过敏原攻击后24 h，支气管内pDCs中GRB的表达上调，并伴随着支气管肺泡灌洗液中GRB浓度的升高。结论我们报道了GRB在人pDC中的选择性表达，并首次证实了PDC介导的GRB和caspase依赖的细胞毒作用。此外，在体外和体内研究了GRB的表达调节，为PDC来源的GRB在过敏性炎症中的特定作用提供了证据。引用如下：K.Bratke，J.Nielsen，F.Manig，C.Klein，M.Kuepper，S.Geyer，P.Julius，M.Lommatzsch和J.C.Virchow，临床与实验过敏，2010(40)1015-1024。
P>BackgroundPlasmacytoid dendritic cells (pDCs) are involved in a variety of immune functions. However, the expression of cytotoxic granule proteins like granzymes and perforin in human pDCs is still poorly understood.ObjectiveThe aim of this study was to systematically analyse the expression and regulation of cytotoxic granule proteins in human pDCs.MethodsThe expression of cytotoxic proteins was analysed by RT-PCR, flow cytometry, and fluorescence microscopy. The functional expression of these proteins was confirmed in a flow-cytometry-based cytotoxicity assay using K562 cells as targets. In order to analyse the regulation of pDC-derived cytotoxic proteins in infectious and allergic diseases, human pDCs were analysed after stimulation with toll-like receptor (TLR)7/9 ligands and in the human asthma model of segmental allergen challenge.ResultsGranzyme B (GrB), but not the granzymes A, H, K, M or perforin, was specifically expressed by human pDCs and this GrB expression was up-regulated by IL-3 stimulation. In addition, IL-3-stimulated pDCs were found to kill K562 cells in a GrB- and caspase-dependent manner. TLR7/9 ligands significantly suppressed GrB expression in pDCs. In contrast, there was an up-regulation of GrB in endobronchial pDCs 24 h after allergen challenge, and this was accompanied by enhanced GrB concentrations in bronchoalveolar lavage fluid.ConclusionWe report the selective expression of GrB in human pDCs and show for the first time pDC-mediated GrB- and caspase-dependent cytotoxicity against target cells. In addition, the regulation of GrB expression was investigated in vitro and in vivo providing an evidence for a specific role of pDC-derived GrB in allergic inflammation.Cite this as: K. Bratke, J. Nielsen, F. Manig, C. Klein, M. Kuepper, S. Geyer, P. Julius, M. Lommatzsch and J. C. Virchow, Clinical & Experimental Allergy, 2010 (40) 1015-1024.