The purpose ofthe Virology and Technology Core is two fold: 1) to provide facilities and technical assistance for the conduction of virus infections and assays in mice, and 2) to facilitate the use of novel assays that we have developed by each ofthe Projects to examine immune responses against alloantigens. One room within a Biosafety level two/three (BSL2/3) Biocontainment Suite will be assigned for work with viruses, for their subsequent surveillance and monitoring, and for the bleeding of mice and harvesting of tissues. For viral and immunological assays, I have a BSL2 laboratory area that has a separate attached tissue culture room equipped with incubators and laminar flow hoods for tissue culture work. This Core will be responsible for the growth, preparation, purification, and assay of viruses, maintaining stocks of peptide epitopes, and for the preparation of antigen-specific MHC-multimer reagents (MHC-lg-dimers and MHCtetramers). The Core will provide reagents and training for assays to quantify and characterize virus-specific T cells including the use of MHC-multimers, T cell proliferation assays, intracellular cytokine assays, and antiviral CTL assays. The new technology to be used include in vivo cytotoxicity assays to measure NK and T cell activity against allogeneic tissues, and intracellular cytokine assays to examine naive, effector/memory, and tolerized alloreactive T cells. The services to be provided by the Core will be available to all Project Laboratories, and the Core will also be responsible for training personnel to perform assays.
The Core will provide essential handling and training in the use of dangerous infectious agents that riequire specialized approaches for safe handling. These viruses have been associated with human disease, and have been found to be detrimental to survival of grafts in the clinic. The availability of these viruses will permit their effect on graft survival using newly developed drugs to be studied in appropriate animal models.
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