Characterisation and quantification of equine interferon gamma.

This study focuses on developing a method to measure the production of Interferon-gamma (IFN-gamma), a crucial element in cell-mediated immunity in horses. The researchers created a technique known as a quantitative ELISA, using monoclonal antibodies produced against the equine variant of IFN-gamma.

Development of the ELISA

• The researchers developed a quantitative enzyme-linked immunosorbent assay (ELISA), which is a test that measures immune responses in the body, specifically for equine lymphocytes creating IFN-gamma.
• They synthesised monoclonal antibodies against the equine variant of IFN-gamma in bacteria, which essentially means that they produced identical immune cells that are all capable of binding to the same antigen.

Validation of the Antibody

• Validation of the monoclonal antibody was carried out using ELISA, Western blotting, flow cytometric and microscopic analysis methods. The antibody proved to be effective at identifying both recombinant and lymphocyte-derived equine IFN-gamma.
• Recombinant refers to the openly structured genetic material in their research, whereas lymphocyte-derived IFN-gamma is the result of certain cells in the immune system.

Observations on Biological Activity

• The team noticed that IFN-gamma deriving from mammalian and insect cells was biologically active, meaning that it maintained metabolic processes and interactions.
• However, one of the monoclonal antibodies was able to neutralise this activity, demonstrating its effectiveness in managing immune responses.

Role of Glycosylation in antiviral activity of eqIFN-gamma

• Surprisingly, the team discovered that glycosylation, the process in which a carbohydrate is added to a protein, seemed to be required to maintain the antiviral properties of equine IFN-gamma.
• This finding could have significant implications for research into equine diseases, as it suggests a potential target for therapeutic intervention.