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Development and Optimisation of a Multiplex Cytokine Assay in Fish.  

Antibody and cell-mediated immune responses are known to be important for disease resistance in mammals. Whilst antibody responses are well established in fish very little is known about cell-mediated immune responses. If such responses are crucial for resistance to certain diseases, then it is important that methods to detect these responses are established. These methods could then be used in the aquaculture industry to monitor the immune status of stocks of fish.

Cell-mediated immunity is an immune response that does not involve antibodies or complement but rather involves the activation of macrophages, natural killer cells (NK), antigen-specific cytotoxic T-lymphocytes, and the release of various cytokines in response to an antigen. It is the cytokines that will be targeted in this project.

Cytokines are critical to the development and functioning of both the innate and adaptive immune response and they are often secreted by immune cells that have encountered a pathogen, thereby activating and recruiting further immune cells to increase the system's response to the pathogen. Interferons and interleukins are molecules classified as cytokines.

The project aims to develop novel assays to measure up and down regulation of cytokines involved in the cellular immune response of fish to pathogens. The assays will rely on using specific antibody probes to detect the cytokine molecules involved and they will use bead array technology to enable measurement of several molecules simultaneously. The bead array system allows for the simultaneous analysis of up to 100 different biomolecules.