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Anti‐ovine VCAM‐1 monoclonal antibodies inhibit adhesion and proliferation between sheep endothelial and mononuclear cells in vitro
Author(s) -
Grooby Warwick L,
Krishnan Ravi,
Russ Graeme R
Publication year - 1997
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1038/icb.1997.85
Subject(s) - monoclonal antibody , vcam 1 , peripheral blood mononuclear cell , in vitro , adhesion , microbiology and biotechnology , antibody , cell adhesion molecule , chemistry , biology , immunology , icam 1 , biochemistry , organic chemistry
This paper reports the production and characterization of three monoclonal antibodies (mAb) recognizing ovine vascular cell adhesion molecule‐1 (VCAM‐1). The mAb were raised against sheep umbilical vein endothelial cells (ShUVEC) and flow cytometric analysis demonstrated that one mAb, QE4G9, was crossreactive with human VCAM‐I expressed on Chinese hamster ovary cell transfectants. Protein modulation studies on ShUVEC and immunoperoxidase staining of inflamed renal tissue further indicated the reactivity of the other two ovine mAb. QE1E3 and QE2G4, with ovine VCAM‐1. The flow cytometric profile of the three mAb on stimulated ShUVEC was identical to that observed with the cross‐reactive anti‐human VCAM‐1 mAb, HAE2‐1. Peak expression occurred between 6–12 h after stimulation, followed by a slight decrease to a plateau extending beyond 48 h. In functional assays, all mAb inhibited adhesion of PMA‐activated sheep PBMC to stimulated ShUVEC. In addition, QE4G9 inhibited proliferation of sheep PBMC in the mixed lymphocyteendothelial cell reaction (MLER) by 56%. The results demonstrate that the three anti‐ovine VCAM‐1 mAb recognize functional epitopes on sheep vascular endothelial cells. These mAb will be valuable tools in the investigation of VCAM‐1 expression in various pathophysiological conditions using sheep models, and in the study of VCAM‐1‐mediated leucocyte‐endothelial cell interactions, both in vitro and in vivo.