Behavior and function of mouse Langerin^+ cells in skin and mucosa.

小鼠皮肤和粘膜中 Langerin^ 细胞的行为和功能。

• 批准号: 13670213
• 负责人: TAKAHARA Kazuhiko
• 金额: $ 2.3万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670213/
• 关键词: dendritic cells; Langerhans cells; Langerin; C-type lectin; FISH; mannan; TGF-β; zymosan; 多重体

Langerin is a C-type lectin that expresses in epidermal dendritic cells, called Langerhans cells. In this study, we cloned mouse Langerin (mLangerin) to investigate its expression pattern and function precisely using mouse system.Based on human Langerin (hLangerin) sequence, cDNA clone that encoded a polypeptid with 65% similarity with hLangerin was cloned. In the result of FISH analysis, the gene was mapped on a syntenic region with that of hLangerin. Additionally, exon-intron structures of h- and mLangerin were conserved well.In skin, expression of mLangerin mRNA was not restricted in epidermis unexpectedly, although protein expression of mLangerin was only observed in epidermal MHC class II^+ cells. mLangerin mRNA as well as protein was also detected in various organs including subcutaneous lymph nodes, spleen, thymus and liver. In suspension cells of spleen, CD11c^+ cells exclusively expressed mLangerin mRNA. Furthermore, CD8α^<high>CD11b^<low> fractions in liver, spleen and subcutaneous lymph nodes expressed predominantly.On the other hand, we found heptad repeats of hydrophobic amino acids in mLangerin neck domain. So intermolecular multimer formation was checked using protein cross-linker. In the result, dimmer and tetramer formation of mLangerin protein was observed and this multimer formation possibly increases avidity of mLangerin for its ligand(s). We next checked binding activity of mLangerin with several sugars. In the result, mLangerin could bind mannan that often exists in microbe cell wall. Using mLangerin transfectant, it was confirmed that mLangerin was involved in capturing of dextran, ovalbumin and zymosan.These results suggest that mLangerin might work as microbe and antigen receptors of CD8α^<high>CD11b^<low> subset of mouse dendritic cells.

Langerin 是一种 C 型凝集素，在表皮树突细胞（称为朗格汉斯细胞）中表达。在本研究中，我们克隆了小鼠Langerin（mLangerin），以利用小鼠系统精确研究其表达模式和功能。基于人Langerin（hLangerin）序列，克隆了编码与hLangerin有65％相似性的多肽的cDNA克隆。 FISH分析结果显示，该基因与hLangerin位于同线性区域。此外，h-和mLangerin的外显子-内含子结构也得到了很好的保存。在皮肤中，mLangerin mRNA的表达出人意料地并未限制在表皮中，尽管mLangerin的蛋白表达仅在表皮MHC II^+细胞中观察到。在皮下淋巴结、脾脏、胸腺和肝脏等多个器官中也检测到 mLangerin mRNA 和蛋白质。在脾脏悬浮细胞中，CD11c^+细胞专门表达mLangerin mRNA。此外，CD8α^<high>CD11b^<low>部分在肝、脾和皮下淋巴结中占主导地位。另一方面，我们在mLangerin颈域中发现了疏水性氨基酸的七联重复序列。因此，使用蛋白质交联剂检查分子间多聚体的形成。结果，观察到 mLangerin 蛋白的二聚体和四聚体形成，并且这种多聚体形成可能增加 mLangerin 对其配体的亲合力。接下来我们检查了 mLangerin 与几种糖的结合活性。结果，mLangerin 可以结合微生物细胞壁中经常存在的甘露聚糖。使用mLangerin转染子，证实mLangerin参与了葡聚糖、卵清蛋白和酵母聚糖的捕获。这些结果表明mLangerin可能作为小鼠树突状细胞CD8α^<high>CD11b^<low>子集的微生物和抗原受体。

项目成果

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Wang, W.-M.：“转化生长因子-β 诱导激活的 ADAMTS-2 的分泌：一种前胶原 III N-蛋白酶”J. Biol. 278. 19549-19557 (2003)

Wang, W. -M., Lee, S., Steiglitz, B. M., Scott, I. C., Lebares, C. C., Allen, L., Brenner, M. C., Takahara, K. and Greenspan, D. S.: "(2003) Transforming growth factor-β induces secretion of activated ADAMTS-2: A procollagen III N-proteinase"J. Biol. Chem

Wang, W. -M.、Lee, S.、Steiglitz, B. M.、Scott, I. C.、Lebares, C. C.、Allen, L.、Brenner, M. C.、Takahara, K. 和 Greenspan, D. S.：“(2003) 转化生长因子-β 诱导激活的 ADAMTS-2 的分泌：前胶原 III N-蛋白酶“J. Biol. Chem

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Kang，Y.-S.：“SIGN-R1，一种由脾边缘区巨噬细胞表达的新型 C 型凝集素，介导多糖葡聚糖的摄取”Int。

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