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A monoclonal antibody disrupting cell-cell adhesion of rat ascites hepatoma cells
Authors:Shigeki Ohshima  Yasuji Ishimaru  Mitsuo Honda  Susumu Ohkawara  Michio Ogawa
Affiliation:(1) Second Department of Surgery, Kumamoto University School of Medicine, Kumamoto, Japan;(2) Department of Surgical Pathology, Kumamoto University School of Medicine, 2-2-1 Honjo, 860 Kumamoto, Japan;(3) Department of Cellular Immunology, National Institute of Health, Tokyo, Japan;(4) First Department of Pathology, Kumamoto University School of Medicine, Kumamoto, Japan
Abstract:
A cell surface-associated adhesive factor (AF) separated from differentiated rat ascites hepatoma AH136B cells (forming cell islands in vivo) has been highly purified by chromatography. AF is assumed to mediate the cell-cell adhesion essential to island formation of the hepatoma cells. A substance, immunologically crossreactive with AF, is present in the ascites fluid or culture medium of the AH136B cells. Because the substance is almost identical to AF in molecular weight and aggregation-promoting activity, it has been concluded that AF is released into the ascites fluid where it is concentrated. Monoclonal antibodies have been raised against AF purified from ascites fluid of AH136B cells. We have obtained a monoclonal antibody, coded MoAF-6D6, that strongly abolishes the aggregation-promoting activity of AF. When AH136B cell islands are incubated in the presence of Fab fragments of MoAF-6D6, cell detachment from the islands is evident within 24 h. Cell islands following 36-h culture show a distinct dissociation and islands completely lose their organization 48 h after culture. The dissociating effect of MoAF-6D6 is neutralized by the addition of AF. These results suggest that AF plays a significant role in the maintenance of cell islands.
Keywords:Ascites hepatoma  Adhesive factor  Monoclonal antibody  Cell membrane polarity  Rat
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