The object of this thesis was to develop and characterise monoclonal antibodies which would recognise damaged and or regenerating endothelial cells and which might therefore be useful in the diagnosis and treatment of vascular disease. The work described was carried out with cultured vascular endothelial cells of human, bovine, or porcine origin. In vitro models of endothelial damage were devised in which cells were damaged by various methods. Mouse monoclonal antibodies were raised by inoculation with cultured endothelial cells and tested against damaged endothelium in vitro. A number of antibody producing clones were identified, isolated and grown on for characterisation of the antibody produced. Using an immunohistochemical system for localising the sites of binding of these antibodies to cultured endothelial cells, a number of different patterns of antibody binding were identified. Attention was concentrated on antibodies which appeared to selectively bind to damaged endothelial cells. In an attempt to identify the antigens recognised by the monoclonal antibodies the protein constituents of endothelial cell lysates were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The protein bands were transferred to nitrocellulose membranes so that specific bands which reacted with individual protein bands could be identified. A number of protein bands were identified which reacted with specific monoclonals. An interaction between damaged endothelial cells and the Fc component of IgG have been previously described in the literature. The non-specific interaction of IgG with endothelial cells was confirmed in this study and the binding component identified. Reasons for believing that the monoclonal antibodies raised in this project predominantly interacted with damaged endothelial components by specific rather than non-specific binding are discussed.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:660768 |
| Date | January 1990 |
| Creators | Pringle, Serena W. |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/20122 |
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