The participation of the host in eliminating Ag-specific T hybridoma cells after their in vivo activation was studied. In our model system, treatment of the cytochrome c-specific T cell hybridoma 2B4.11 in vitro with Ag in the context of histocompatible APC results in cellular activation, as shown by IL-2 release and growth inhibition. In vivo treatment with Ag results in tumor cell elimination as a result both of a direct inhibitory effect of cytochrome c that is mediated through the 2B4.11 TCR and to the induction of host immunity. In vivo lymphocyte-depletion studies showed that CD8-bearing cells were critical to the successful elimination of tumor cells mediated by Ag, whereas depletion of CD4-bearing cells had only minor effects on the outcome. Cytotoxic cells from mice cured by Ag treatment lysed only 2B4.11 among a panel of related tumors, although in vivo cross-protection studies showed that 2B4.11-immune mice were also resistant to the growth of BW5147 and C10.9. Because spleen cells from 2B4.11 immune mice did not recognize 2B4.11 or other related tumors in proliferation assays, we concluded that a participant(s) with memory and specificity, not assayed in vitro, was also involved in the mediation of the immune effects observed. For therapies based on the use of less selective agents, i.e. mAb that share the activating properties of Ag but can react with T cell neoplasms of unknown specificity, it would appear that a relatively intact immune system is required for maximal success.