A unique monoclonal antibody mNI-11 rapidly enhances spread formation in human umbilical vein endothelial cells.
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.
Division of Immunology, Kyushu University of Health and Welfare, Faculty of Health and Science, Nobeoka-city, Japan.
N
N
Ikewaki
H
H
Tamauchi
A
A
Yamada
N
N
Mori
H
H
Yamao
H
H
Inoue
H
H
Inoko
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D000918:Antibody Specificity; D002118:Calcium / Q000502:physiology; D002147:Calmodulin / Q000037:antagonists & inhibitors; D015815:Cell Adhesion Molecules / Q000276:immunology* / Q000502:physiology; D048429:Cell Size; D002478:Cells, Cultured / Q000187:drug effects / Q000648:ultrastructure; D002614:Chelating Agents / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D019833:Genistein / Q000494:pharmacology; D018922:HL-60 Cells / Q000187:drug effects / Q000648:ultrastructure; D006801:Humans; D020014:K562 Cells / Q000187:drug effects / Q000648:ultrastructure; D008870:Microtubules / Q000187:drug effects; D015739:Nocodazole / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology; D013449:Sulfonamides / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D020298:U937 Cells / Q000187:drug effects / Q000648:ultrastructure; D014471:Umbilical Veins
D000911:Antibodies, Monoclonal; D002147:Calmodulin; D015815:Cell Adhesion Molecules; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D000939:Epitopes; D011500:Protein Synthesis Inhibitors; D013449:Sulfonamides; D014409:Tumor Necrosis Factor-alpha; D015638:Cytochalasin D; C017967:W 7; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D003513:Cycloheximide; D004492:Edetic Acid; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D015739:Nocodazole; D002118:Calcium