ENDOTHELIAL LEUKOCYTE ADHESION MOLECULE-1 - DIRECT EXPRESSION CLONING AND FUNCTIONAL INTERACTIONS
ENDOTHELIAL LEUKOCYTE ADHESION MOLECULE-1 - DIRECT EXPRESSION CLONING AND FUNCTIONAL INTERACTIONS
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DOI：
10.1073/pnas.87.5.1673
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发表时间：
1990-03-01
影响因子：
11.1
通讯作者：
LOBB, R
中科院分区：
文献类型：
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作者：
HESSION, C;OSBORN, L;LOBB, R
用细胞因子处理的人脐静脉内皮细胞(HUVECs)的消减文库在COS-7细胞中瞬时表达内皮细胞白细胞黏附分子-1(ELAM-1)，并通过与人早幼粒细胞系HL-60的黏附筛选获得表达ELAM-1的克隆。这种克隆方法既不需要抗体，也不需要纯化的配体。表达ELAM-1的COS细胞与早幼粒细胞系HL-60的结合依赖于钙离子，但不依赖于温度。虽然ELAM-1与哺乳动物凝集素同源，但它与HL-60细胞的相互作用不受简单的碳水化合物结构的抑制。表达ELAM-1的COS细胞还可以与人中性粒细胞和人结肠癌细胞株HT-29结合，但不能与B细胞系Ramos结合。然而，Ramos细胞与细胞因子处理的HUVECs黏附，但不控制HUVECs，证实了其他可诱导黏附分子的存在。此外，针对CD18抑制性表位的单抗(60.3)不能抑制HL-60细胞或中性粒细胞与表达ELAM-1的COS细胞的结合，这表明ELAM-1配体虽然未被鉴定，但不是CD11/CD18家族的成员。
A cDNA for endothelial leukocyte adhesion molecule 1 (ELAM-1) was isolated by transient expression in COS-7 cells of a subtracted cDNA library from cytokine-treated human umbilical vein endothelial cells (HUVECs), with selection of ELAM-1-expressing clones by adhesion of transfected cells to the human promyelocytic cell line HL-60. This cloning method requires neither antibody nor purified ligand. ELAM-1-expressing COS cells bind the promyelocytic cell line HL-60 by a Ca2+ -dependent but temperature-independent mechanism. Although ELAM-1 is homologous to mammalian lectins, its interaction with HL-60 cells is not inhibited by simple carbohydrate structures. ELAM-1-expressing COS cells also bind-human neutrophils and the human colon carcinoma cell line HT-29, but not the B-cell line Ramos. However, Ramos cells adhere to cytokine-treated HUVECs but not control HUVECs, confirming the existence of other inducible adhesion molecules. In addition, the binding of HL-60 cells or neutrophils to ELAM-1-expressing COS cells is not inhibited by a monoclonal antibody (60.3) directed to an inhibitory epitope on CD18, indicating that the ELAM-1 ligand, although uncharacterized, is not a member of the CD11/CD18 family.