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Title: Studies on ovine interferon-gamma
Author: Henfrey, Andrew Mark
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1993
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Interferons have been recognized as important mediators of cellular communication for many years. There are two types of interferon: Type I interferons have antiviral functions, but Type II interferon (IFN-g) is more important as an immunomodulating molecule. Type II interferon has effects on cellular MHC class II expression, immunoglobulin class-switching, macrophage activation, cellular proliferation and a number of other functions. The role of IFN-g during in vivo immune responses has not been studied in great detail, but the sheep is an ideal species in which to study these phenomena by using the efferent lymphatic vessel cannulation model. This allows access to cells and tissue fluid for cytokine analysis using antibody and genetic probes for the detection of IFN-g. Bovine IFN-g peptides (amino-terminus, carboxy-terminus and central) were used to generate antibodies in rabbits. None of the anti-peptide sera reacted with denatured ovine or bovine IFN-g, nor neutralized their antiviral effect. Rabbit antibodies to bovine recombinant IFN-g neutralized ovine IFN-g and detected IFN-g in a sandwich ELISA when used in combination with a monoclonal antibody against a human IFN-g carboxy-terminal peptide. The sensitivity of detection was only 125ng/ml, insufficient for use with efferent lymph fluid samples. The expression of MHC class II molecules on cell surfaces is increased by IFN-g on many cell types. This has been used previously to measure biologically active IFN-g concentrations in fluids. Measurement of ovine class II by slot blot was assessed as a method of adapting this to ovine IFN-g measurement, but the technique proved to be too problematic for regular use. The expression of class II on T lymphocytes is influenced by IFN-g in the surrounding fluid. Analysis by FACS of resting ovine T lymphocytes shows them to express class II, a situation different to that in the human.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available