The goal of the proposed study is to obtain training and experience in the biochemical, immunological, and genetic characterization of specific products coded for by the major histocompatability complex (MHC) of the dog. The interdisciplinary approach, involving biochemistry, immunology, transplantation biology, hybridoma biotechnology, and canine reproduction, will provide the opportunity for a broadened research environment which can be used to focus on attempting to answer questions regarding the significance of the MHC in immunobiology. Specific laboratory techniques will be geared towards learning the methodology and background of developing new alloantisera, monoclonalantisera, and homozygous and primed typing lymphocytes. Experience in breeding, semen cryopreservation, and artifical insemination will enable maintenance and expansion of a breeding colony of reference donor dogs homozygous for MHC determinants. The objective is to produce and expand a comprehensive typing panel which can be used to study functional in vitro aspects of MHC products. This will include (1) correlating monoclonal antisera with currently (and possibly new) defined alloantisera; (2) determining the relationship between reactive lymphocytes from homozygous dogs, primed lymphocytes, and allo and/or monoclonal antisera in the mixed lymphocyte reaction and primed lymphocyte tests; and (3) defining the locus (or loci) involved in coding for primed lymphocyte reactivity. A second objective is to use an expanded typing panel to more accurately select allogeneic donors for canine bone marrow transplantation studies conducted at FHCRC. The third objective is to provide a resource for studying the role of MHC products in transplantation biology and the association with specific canine diseases, especially those serving as models for human disease. The potential would exist for collaborative studies not only at the institutional level but also on a national and international level by selective panel typing or the exchange of antisera, typing cells, semen, or dogs.
