The soluble fraction from a sonicate of Toxoplasma gondii tachyzoites (termed F3) was shown to induce dose-dependent blastic transformation of peripheral-blood mononuclear cells (PBMC) from seropositive individuals only and was used to isolate a panel of T-cell clones from the PBMC of an immune donor. Proliferation assays using F3 showed that 15 (14 CD4+ and 1 CD8+) of the 18 isolated clones were specific for T. gondii. In response to antigen stimulation, 5 of the 15 clones produced detectable levels of interleukin-2 (IL-2, 0.2-15 u/ml) and 9 clones produced significant levels of interferon-gamma (IFN-gamma, 17.5-1400 IU/ml). Seven of the 7 T-cell clones tested reacted with two different Toxoplasma strains (RH and Wiktor). When used as antigen-presenting cells, an autologous B-lymphoblastoid cell line could efficiently present the antigen to only three of the six T-cell clones tested. This study identifies and characterizes cellular probes that could be useful for future vaccine design.