A unique monoclonal antibody mNI-11 rapidly enhances spread formation in human umbilical vein endothelial cells.
J Clin Immunol, 2000/7;20(4):317-24.
Ikewaki N[1], Tamauchi H, Yamada A, Mori N, Yamao H, Inoue H, Inoko H
Affiliations
PMID: 10939719
Impact factor: 8.542
Abstract
We previously reported a novel monoclonal antibody (MAb), designated mNI-11, recognizing an adhesion-associated antigen distinct from any previously reported ones. In this article, this adhesion-associated antigen with a molecular weight of about 97 kDa was found to be strongly expressed on human umbilical vein endothelial cells (HUVECs) by fluorescence-activated cell sorter (FACS) analysis. Expression of this antigen on HUVECs was slightly increased in response to the exposure to tumor necrosis factor-alpha (TNF-alpha) or phorbol myristate acetate (PMA). As a biological function exerted by this antigen, it was of great interest that immobilized mNI-11 directly and rapidly enhanced the spread formation of HUVECs, whereas MAbs binding other adhesion-associated antigens such as mNI-58A (anti-CD11a), L130 (anti-CD18), L133.1 (anti-CD31), L178 (anti-CD44), L25.3 (anti-CD49d), or LB-2 (anti-CD54) did not carry such activity under the same conditions. The HUVECs spread formation enhanced by mNI-11 was completely blocked in the presence of a microfilament formation inhibitor, cytochalasin D (CyD), a Ca2+ calmodulin inhibitor, W-7, EDTA, and was partially blocked by a microtubule formation inhibitor, nocodazole, a protein kinase C (PKC) inhibitor, H-7, and a protein synthesis inhibitor, cycloheximide (CHX). However, a protein tyrosine kinase (PTK) inhibitor, genistein, did not affect the spread formation under the same conditions. Taken together, it was suggested that the spread formation of HUVECs enhanced by mNI-11 was mainly associated with the influx of Ca2+ and microfilament reorganization. In addition, the novel property associated with mNI-11 to enhance the spread formation of HUVECs was possibly mediated through its reaction against a unique epitope on HUVECs.
MeSH terms
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Actin Cytoskeleton; Antibodies, Monoclonal; Antibody Specificity; Calcium; Calmodulin; Cell Adhesion Molecules; Cell Size; Cells, Cultured; Chelating Agents; Cycloheximide; Cytochalasin D; Edetic Acid; Endothelium, Vascular; Enzyme Inhibitors; Epitopes; Flow Cytometry; Genistein; HL-60 Cells; Humans; K562 Cells; Microtubules; Nocodazole; Protein Kinase C; Protein Synthesis Inhibitors; Protein-Tyrosine Kinases; Sulfonamides; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor-alpha; U937 Cells; Umbilical Veins
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