Translocation of active heparanase to cell surface regulates degradation of extracellular matrix heparan sulfate upon transmigration of mature monocyte-derived dendritic cells
Translocation of active heparanase to cell surface regulates degradation of extracellular matrix heparan sulfate upon transmigration of mature monocyte-derived dendritic cells
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DOI：
10.4049/jimmunol.176.11.6417
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发表时间：
2006-06-01
影响因子：
4.4
通讯作者：
Mevorach, Dror
中科院分区：
文献类型：
--
作者：
Benhamron, Sandrine;Nechushtan, Hovav;Mevorach, Dror
在Ag捕获和暴露于危险刺激后，成熟的树突状细胞（DC）迁移到局部淋巴结，在那里抗原肽呈递给T淋巴细胞。为了从外周组织如表皮迁移到区域淋巴结，载银表皮朗格汉斯细胞必须移动通过各种组成的细胞外基质（ECM）。尽管MIP-3 β和CCR 7起着关键作用，但它们通过ECM迁移的能力的性质还不清楚。我们感兴趣的是验证乙酰肝素酶，一种参与ECM降解和重塑的硫酸乙酰肝素降解内切-β-D-葡萄糖醛酸酶，是否在单核细胞来源的DC中表达和发挥功能。使用免疫组织化学、共聚焦显微镜、RT-PCR、Western印迹分析、乙酰肝素酶活性测定和Matrigel迁移，我们表明乙酰肝素酶在未成熟DC的细胞核和细胞质中表达，并且基因表达和合成主要发生在单核细胞和早期未成熟MS中。我们还发现，细胞核和细胞质组分均显示乙酰肝素酶活性，并且在LPS诱导的成熟中，乙酰肝素酶易位至细胞表面并降解ECM硫酸乙酰肝素。基质胶迁移试验显示，类肝素酶的MIP-3 β-相当的作用。由于硫酸乙酰肝素糖胺聚糖在ECM的自组装、不溶性和屏障特性中起关键作用，因此本研究的结果表明乙酰肝素酶是DC通过ECM迁移的关键酶。
After Ag capture and exposure to danger stimuli, maturing dendritic cells (DCs) migrate to regional lymph nodes, where antigenic peptides are presented to T lymphocytes. To migrate from peripheral tissue such as the epidermis to regional lymph nodes, Ag-bearing epidermal Langerhans cells must move through an extracellular matrix (ECM) of various compositions. The nature of their capacity to transmigrate via ECM is not well understood, although MIP-3 beta and CCR7 play critical roles. We were interested in verifying whether heparanase, a heparan sulfate-degrading endo-beta-D-glucuronidase that participates in ECM degradation and remodeling, is expressed and functional in monocyte-derived DCs. Using immunohistochemistry, confocal microscopy, RT-PCR, Western blot analysis, assays for heparanase activity, and Matrigel transmigration, we show that heparanase is expressed in both nuclei and cytoplasm of immature DCs, and that gene expression and synthesis take place mainly in monocytes and early immature Ms. We also found that both nuclear and cytoplasm fractions show heparanase activity, and upon LPS-induced maturation, heparanase translocates to the cell surface and degrades ECM heparan sulfate. Matrigel transmigration assays showed a MIP-3 beta-comparable role for heparanase. Because heparan sulfate glycosaminoglycans play a key role in the self-assembly, insolubility, and barrier properties of the ECM, the results of this study suggest that heparanase is a key enzyme in DC transmigration through the ECM.