Immune tolerance to MHC class II identical renal grafts is achievable in miniature swine following a short immunosuppressive treatment. Like in clinical transplants, swine-accepted allografts are primarily infiltrated by CD8(+) T cells, which are noncytotoxic to the renal tissue. However, the actual specificity and function of these intragraft-infiltrating lymphocytes remain poorly understood. To develop the molecular tools to study TCR-associated functions of graft-infiltrating cells in a preclinical transplantation model, we have determined the nucleotide sequence of 19 pig Vbeta, 12 Jbeta, and two Dbeta. Sequence comparisons identified 17 different Vbeta families and two Jbeta clusters homologous to the human Jbeta1 and Jbeta2. The fact that the pig Jbeta1 segments were always found joined to the Dbeta1-like sequence in numerous rearranged TCR beta cDNA suggests the existence of two D-J clusters in swine. These results also imply that the polymorphism of the porcine TCR beta segments is similar to that found in human. Finally, we report the discovery of a new and functional Vbeta subfamily named Vbeta100, which exhibited similarity to the murine Vbeta2 sequence but had no described Vbeta homolog in humans. Pilot spectratyping studies on Vbeta usage revealed a clonal dominance of Vbeta100(+) T cell subsets among infiltrating cells in two accepted grafts.