Development of cytotoxicity test for outbred animals using recombinant viruses.

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The cytotoxic T lymphocytes (CTL) have been considered to play an important role in immunity against the infections of many kinds of viruses. However, the function of CTL in animals including domestic, companion or experimental animals except mice were not well studied because of the lack of CTL assay methods for those animals. It was difficult to produce the useful target cells in those animal systems since target cells should be obtained from the same animal which is infected by viruses to match MHC with the effector cells, and since the target cells should express virus antigens on the cell surface without showing CPE until they meet the effector cells.The final aim of this project is to develop and establish the new cytotoxic T cell assay system which is useful for any kinds of animals by using improved adenovirus vectors. For developing the new assay system, we tested and obtained the optimal conditions in each subjects as follows.(1) Host animal and cell ranges of adenovirus infection.(2) The best promoters to express virus antigens in the adenovirus vector.(3) Cloning and characterizations of virus genes whose products are considered to be the target of cellular cytotoxicity.(4) Construction of recombinant adenoviruses with virus genes.(5) The conditions to produce the target cells from peripheral lymphocytes of each animals after the infection of recombinant adenoviruses.Since we tested several kinds of viruses, not all those conditions have been obtained in each system. But we succeeded to construct recombinant adenoviruses with measles and renderpest virus genes, and obtained the optimal conditions for producing the target cells from monkey lymphocytes infected with the recombinant adenoviruses. In addition, we identified the best promoters in adenovirus vectors, and the unique MHC molecules on bovine lymphocytes. These results are considered to contribute to the basic and applied studies for cellular cytotoxicity of animals using adenovirus vectors.