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ObjectiveWith the aim of obtaining monoclonal antibodies (mAbs) against mouse endothelial surface antigens, immunization of rats with a mouse-derived endothelial cell line (PY4.1) and subsequent hybridoma production were performed.Materials and methodsOne of the mAbs produced by hybridoma EOL5F5 was selected for its surface binding to endothelial cell lines, and identification of the mAb-recognized antigen was performed by immunoprecipitation. Experiments were performed to analyze the effects of EOL5F5 on systemic administration to mice.ResultsEOL5F5-recognized antigen was a single band of 35 kDa under reducing and nonreducing conditions, features that do not match other known differentiation antigens with comparable tissue distribution. In vivo administration of purified EOL5F5 mAb to mice (n = 20) induced intense cutaneous purpura as well as severe but transient thrombocytopenia. Expression of EOL5F5-recognized antigen was detected on platelets from which it immunoprecipitated a moiety of identical electrophoretic pattern in SDS-PAGE, as the one recognized on endothelial cells. Immunohistochemically, EOL5F5-recognized antigen (p35) also was expressed on dermal capillaries, suggesting that, in addition to thrombocytopenia, damaging effects of the antibody on endothelial cells also might cause the observed purpura.ConclusionsOur results show induction of thrombocytopenic purpura in mice with an mAb against a single antigenic determinant expressed on both platelets and endothelium. EOL5F5 mAb injection sets the stage for useful experimental models that resemble immune thrombocytopenic purpura.

Original publication

DOI

10.1016/s0301-472x(01)00630-0

Type

Journal article

Journal

Experimental hematology

Publication Date

05/2001

Volume

29

Pages

589 - 595

Addresses

Department of Medicine, University of Navarra, Pamplona, Spain.

Keywords

Endothelium, Vascular, Blood Platelets, Hybridomas, Skin, Animals, Mice, Mice, Nude, Rats, Rats, Wistar, Purpura, Thrombocytopenic, Thrombocytopenia, Antigens, Differentiation, Antibodies, Monoclonal, Antigens, Surface, Epitopes, Immunization