Monoclonal antibodies to uveal melanoma

Damato, Bertil E (1987) Monoclonal antibodies to uveal melanoma. PhD thesis, University of Glasgow.

Abstract

This thesis concerns the immunology of uveal melanoma It is introduced by a critical review of the literature on this neoplasm, and on tumour immunology with special reference to melanoma Two main conclusions could be drawn from this review Firstly, further progress in the immunology of uveal melanoma is dependent on a better understanding of the antigenicity of this tumour and on the availability of suitable antigens for testing the anti-tumour immune response Secondly, the best chance of achieving these objectives, at present, is by the application of monoclonal antibody technology Accordingly, the main aims of this study were to determine the feasibility and scope of producing monoclonal antibodies to uveal melanoma Monoclonal antibodies were prepared using the rat hybridoma system Tissue specimens from over 80 uveal melanomas were used to immunise rats and for the necessary immuno-assays Hybridomas were screened by ELISA Eleven monoclonal antibodies were produced Most of these were of the IgM isotype and all reacted with intracellular antigens The most specific was mAb 4A3 Nevertheless, immunohistochemistry on frozen and fixed sections and immunofluorescence microscopy using cell suspensions showed that this antibody reacted with several types of normal and malignant cells Western blotting demonstrated that the 4A3 antigen had a molecular weight of 55-62 kD The 4A3 antigen was detected in the subretinal fluid of five patients with uveal melanoma with the use of this technique, and in one of two patients with rhegmatogenous retinal detachment ELISA reactivity of the rat monoclonal antibodies with uveal melanoma cells was inhibited by serum from 10 out of 12 patients with uveal melanoma and from two out of eight healthy individuals B Lymphocytes of patients with uveal melanoma were EBV transformed in vitro and tested by ELISA for reactivity with autologous tumour The results were similar to those obtained with lymphocytes from healthy individuals This suggested that such techniques were inadequate for analysing humoral immunity to melanoma and unsuitable for the production of human monoclonal antibodies to this tumour In conclusion, specific monoclonal antibodies to uveal melanoma could be very useful, but such antibodies are unlikely to be produced using conventional methods Better results will probably be achieved if uncultured tissue from a single tumour were used for all immunisation and assay procedures required for one fusion, and if more efficient techniques for selecting hybridomas were available.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Keywords: Medicine, Oncology, Immunology
• Date of Award: 1987
• Depositing User: Enlighten Team
• Unique ID: glathesis:1987-76680
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 19 Nov 2019 13:55
• Last Modified: 19 Nov 2019 13:55
• URI: https://theses.gla.ac.uk/id/eprint/76680