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Localization of lymphocyte homing ligands, CD34 and GlyCAM-1

淋巴细胞归巢配体 CD34 和 GlyCAM-1 的定位

基本信息

批准号：
07670016
负责人：
KIKUTA Akio
金额：
$ 1.02万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670016/
关键词：
GlyCAM-1 CD34 homing ligand high endothelial venules L-selectin マウス

项目摘要

L-selectin is a lymphocyte homing receptor and two high endothelial venule (HEV)-associate ligands for L-selectin, known as GlyCAM-1 and CD34, have been identified. Using colloidal gold conjugated-recombinant L-selectin (LSIG), antibodies against CD34 and GlyCAM-1, and an anti-peripheral node addressin mAb (MECA79), we performed morphological mapping of L-selectin ligands in peripheral lymph nodes. For MECA-79 and LSIG,intense labeling was detected on the luminal surface and cytoplasm of the high endothelial cells, and the fibrous layr of the perivascular connective tissue (PVC) of the HEV.The L-selectin ligand in the HEV-PVC,through which extravasated lymphocytes pass into the parenchyma of the lymph node, was found to be neither CD34 nor GlyCAM-1 and another one.In order to measure the changes in secreted plasma GlyCAM-1 levels, we performed sandwich ELISA assay after inflammatory stimulus. BALB/c mice were injected with complete Freund adjuvant in the hind footpads. GlyCAM-1 was increased at 3 h and reached peak levels at 12 h and gradually decreased thereafter. Levels of EGTA-sensitive L-selectin bound to the plasma GlyCAM-1 changed in similar time course and reached peak at 12 h after and began to decrease. These findings show that GlyCAM-1 release is enhanced by inflammatory stimulation and also suggest that released plasma GlyCAM-1 may trap, at least in part, soluble L-selectin shed from stimulatecd leukocytes to neutralize with each other.

L-选择素是一种淋巴细胞归巢受体，目前已鉴定出L-选择素的两种高内皮微静脉相关配体，即GlyCAM-1和CD34。应用胶体金标记的重组L-选择素、抗CD34和GlyCAM-1的抗体以及抗外周淋巴结寻址蛋白的单抗(MECA79)，对L-选择素配体在周围淋巴结中的定位进行了研究。对于MECA-79和LSIG，在HEV的高内皮细胞的管腔表面和胞浆以及血管周围结缔组织(PVC)的纤维层上都有强标记。HEV-PVC中的L-选择素配体既不是CD34也不是GlyCAM-1，而是另一种配体。BALB/c小鼠后足注射完全弗氏佐剂。GlyCAM-1于3h开始升高，12h达高峰，以后逐渐下降。血浆GlyCAM-1结合的EGTA敏感的L-选择素水平变化趋势相似，12h达高峰，随后开始下降。这些结果表明炎症刺激可促进GlyCAM-1的释放，也提示释放的血浆GlyCAM-1可能至少部分捕获了从刺激的白细胞中脱落的可溶性L-选择素，使其相互中和。