This programme aims to take advantage of our recent success in cloning fish cytokines, to develop fish leucocyte cell lines. We will produce the recombinant proteins and add to leucocyte primary cultures as growth factors to allow their growth in vitro. In addition, we will combine this treatment with antigen stimulation, by using leucocytes derived from immunised fish. In addition to morphological studies, we will undertake molecular and functional analysis to give a preliminary characterisation of the cell types that grow under these conditions and determine their cytotoxic and phagocytic activities. We will test whether the cell lines can be used to measure responses to exogenous immunostimuli such as lipopolysaccride, polyI:C and recombinant cytokines. We believe that the established cell lines will provide a permanent platform for basic research of the fish immune system, thereby reducing the need to kill fish to harvest tissues. They will also help reduce fish suffering by allowing some testing of aquaculture drugs/reagents, and the impact of some chemicals/pollutants to be carried out in vitro before in vivo studies, to establish whether they impact on leucocyte function and therefore potentially fish health and welfare.
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