A panel of cytotoxic T lymphocyte clones that recognize H-2b target cells has been established. Six different clones were distinguished according to the following criteria. First, the fine specificity of the clones was determined by testing proliferation and cytotoxicity on target cells of recombinant mice. Clone 221 recognized H-2Kb, and five other clones recognized H-2Db. Clone 433 distinguished itself from the other five Db-specific clones by cross-reacting with an antigen on H-2k cells. Second, the presence of an idiotypic determinant as defined by the 3F9 clone-specific monoclonal antibodies was investigated in cytotoxicity inhibition experiments. One of the Db-specific clones, 653, was inhibited by these antibodies and was therefore clearly different from the other Db-specific clones. The third criterion involved the rearrangement pattern of the DNA coding for the beta chain of the T-cell receptor. Southern blot analysis showed that each clone had a unique pattern. Interestingly, clone 653, which expresses the same idiotypic determinant as clone 3F9, had deleted the C beta 1 gene cluster, whereas this gene is functionally expressed in clone 3F9.