Molecular mechanisms underlying lymphocyte-specific cell trafficking into lymph nodes-Functional analysis of genes specifically expressed in high endothelial venules (HEV)

淋巴细胞特异性细胞转运至淋巴结的分子机制——高内皮微静脉 (HEV) 中特异性表达的基因的功能分析

• 批准号: 11470057
• 负责人: MIYASAKA Masayuki
• 金额: $ 8.51万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470057/
• 关键词: lymphocyte homing; high endothelial venule; chemokine; lymph node; endothelial cell; sulfotransferase; 接着分子; 組織特異的遺伝子; cDNAライブラリー; リンパ球

We have successfully isolated endothelial cells from HEV (HEV cells) of peripheral and mesenteric lymph nodes of mice and constructed HEV-specific 3'-end cDNA libraries. We then randomly sequenced about 1,500 cDNA clones from each library and compared the obtained nucleotide sequences with those found in 30 different cDNA libraries similarly obtained from other tissues and also with those registered in the EST database. From this analysis, we found several molecules selectively expressed by HEV cells but not normal endothelial cells. They include mac25/TAF/PSF, SGP-2, DARC, leucine-rich α2 glycoprotein and Ig-hepta. In addition, a few chemokines such as SLC, ELC, BLC, fractalkine, KC and lymphotactin are expressed by HEV at mRNA and protein levels. These chemokines except KC can attract selective lymphocyte subsets in vitro, and hence are strategically useful for recruiting these subsets into lymph nodes. In addition, in collaboration with Prof.M.Minoru of La Jolla, we have successfully cloned a novel GlcNAc-6-O-sulfotransferase selectively expressed inHEV.This enzyme is essential for the formation of an L-selectiin binding sugar moiety, 6-sulfo sialyl Lewis X, and is likely to play a crucial role in the L-selectin-dependent lymphocyte homing into lymph nodes.

我们成功地从戊型肝炎病毒（HEV）感染的小鼠外周和肠系膜淋巴结中分离出内皮细胞（HEV细胞），并构建了HEV特异性3 '端cDNA文库。然后，我们从每个文库中随机测序了约1，500个cDNA克隆，并将获得的核苷酸序列与类似地从其他组织获得的30个不同cDNA文库中发现的核苷酸序列以及EST数据库中登记的核苷酸序列进行了比较。从这个分析中，我们发现了几个分子选择性表达的戊型肝炎病毒细胞，而不是正常的内皮细胞。它们包括mac 25/TAF/PSF、SGP-2、DARC、富含亮氨酸的α2糖蛋白和Ig-hepta。此外，一些趋化因子如SLC、ELC、BLC、fractalkine、KC和趋化因子在mRNA和蛋白水平上由HEV表达。这些趋化因子，除了KC可以吸引选择性淋巴细胞亚群在体外，因此是战略上有用的招募这些亚群进入淋巴结。此外，与拉霍亚的Minoru教授合作，我们已经成功克隆了一种新的选择性表达于HEV的GlcNAc-6-O-磺基转移酶。这种酶对于形成L-选择素结合糖部分6-磺基唾液酸刘易斯X是必需的，并且可能在L-选择素依赖性淋巴细胞归巢到淋巴结中起关键作用。

项目成果

Pabst,R.: "Lymphocyte migration. More than an interaction of lymphocytes with endothelial cells of the high endothelial venules via adhesion molecules."The Immunologist. 7. 1-2 (1999)

Pabst, R.：“淋巴细胞迁移。不仅仅是淋巴细胞通过粘附分子与高内皮微静脉的内皮细胞相互作用。”免疫学家。

Ding,Z.: "Sulfatide binding and activation of leukocytes through an L-selectin-independent pathway."J.Leuk.Biol.. 68. 65-72 (2000)

Ding,Z.：“通过 L-选择素独立途径实现硫脂结合和白细胞激活。”J.Leuk.Biol.. 68. 65-72 (2000)

Pabst, R.& Miyasaka, M.: "Lymphocyte migration. More than an interaction of lymphocytes with endothelial cells of the high endothelial venules via adhesion molecules."The Immunologist. 7. 1-2 (1999)

帕布斯特，R.

Li,Y.-F.: "Identification and characterization of ligands for L-selectin in the kidney.II.. Expression of chondroitin sulfate and heparan sulfate proteoflycans reactive with L-selectin"FEBS Letters. 444. 201-205 (1999)

Li，Y.-F.：“肾脏中 L-选择素配体的鉴定和表征。II..与 L-选择素反应的硫酸软骨素和硫酸乙酰肝素蛋白聚糖的表达”FEBS Letters。

Izawa,D.: "Expression profile of active genes in mouse lymph node high endothelial cells"Int.Immunol.. 11. 1989-1997 (1999)

Izawa,D.：“小鼠淋巴结高内皮细胞中活性基因的表达谱”Int.Immunol.. 11. 1989-1997 (1999)