Functional Interaction of Common Allergens and a C-type Lectin Receptor, Dendritic Cell-specific ICAM3-grabbing Non-integrin (DC-SIGN), on Human Dendritic Cells
Functional Interaction of Common Allergens and a C-type Lectin Receptor, Dendritic Cell-specific ICAM3-grabbing Non-integrin (DC-SIGN), on Human Dendritic Cells
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DOI：
10.1074/jbc.m109.058370
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发表时间：
2010-03-12
影响因子：
4.8
通讯作者：
Huang, Shau-Ku
中科院分区：
文献类型：
--
作者：
Hsu, Shih-Chang;Chen, Chien-Ho;Huang, Shau-Ku
已知病原体上的岩藻糖基化聚糖通过其与树突细胞（DC）上的模式识别受体（例如C型凝集素受体（CLR））的相互作用来塑造免疫应答。类似的岩藻糖基化结构也常见于各种过敏原中，但其功能意义仍不清楚。为了检验过敏原相关聚糖作为与CLR功能性相互作用的分子模式的假设，进行了基于酶联免疫吸附试验的结合试验，以确定纯化过敏原和过敏原提取物的结合活性。THP-1细胞和单核细胞衍生的DC（MDDC）作为一个模型，用于测试过敏原-β-内酰胺酶相互作用的功能效果，使用酶联免疫吸附试验，蛋白质印迹和流式细胞术进行了研究。发现了与DC特异性ICAM 3抓取非整合素（DCSIGN）及其相关受体L-SIGN具有可变结合活性的过敏原和过敏原提取物的显著和饱和结合。（岩藻糖基化聚糖缺乏α 1，3-连接甘露糖）的过敏原和一组纯化的过敏原，包括BG 60（Cyn dBG-60;百慕大草花粉）和Der p2（屋尘螨）。结合活性是钙依赖性的，并可被岩藻糖和Lewis-x三羟甲基氨基甲烷（Lex）取代。在THP-1细胞和人MDDC中，BG 60-DC-SIGN相互作用导致Raf-1和ERK激酶的激活以及肿瘤坏死因子-α表达的诱导。这种作用可以被Raf-1抑制剂或抗DC-SIGN抗体部分阻断，并且在DC-SIGN敲低的细胞中显著降低。这些结果表明，过敏原能够与DC-SIGN相互作用，并部分地通过Raf-1信号传导途径诱导MDDC中的肿瘤坏死因子-α表达。
Fucosylated glycans on pathogens are known to shape the immune response through their interaction with pattern recognition receptors, such as C-type lectin receptors (CLRs), on dendritic cells (DCs). Similar fucosylated structures are also commonly found in a variety of allergens, but their functional significance remains unclear. To test a hypothesis that allergen-associated glycans serve as the molecular patterns in functional interaction with CLRs, an enzyme-linked immunosorbent assay-based binding assay was performed to determine the binding activity of purified allergens and allergen extracts. THP-1 cells and monocyte-derived DCs (MDDCs) were investigated as a model for testing the functional effects of allergen-CLR interaction using enzyme-linked immunosorbent assay, Western blotting, and flow cytometry. Significant and saturable bindings of allergens and allergen extracts with variable binding activities to DC- specific ICAM3-grabbing non-integrin (DCSIGN) and its related receptor, L-SIGN, were found. These include bovine serum albumin coupled with a common glycoform (fucosylated glycan lacking the alpha 1,3-linked mannose) of allergens and a panel of purified allergens, including BG60 (Cyn dBG-60; Bermuda grass pollen) and Der p2 (house dust mite). The binding activity was calcium-dependent and inhibitable by fucose and Lewis-x trisaccharides (Lex). In THP-1 cells and human MDDCs, BG60-DC-SIGN interaction led to the activation of Raf-1 and ERK kinases and the induction of tumor necrosis factor-alpha expression. This effect could be blocked, in part, by Raf-1 inhibitor or anti-DC-SIGN antibodies and was significantly reduced in cells with DC-SIGN knockdown. These results suggest that allergens are able to interact with DC- SIGN and induce tumor necrosis factor-alpha expression in MDDCs via, in part, Raf-1 signaling pathways.