Peritoneal macrophages (PM) from normal Wistar rats were treated in vitro with peritoneal supernatant or sera, obtained from rats infected with 10(6) Trypanosoma lewisi trypomastigotes before the infection with Toxoplasma gondii tachyzoites. In this experimental model, Toxoplasma multiplication in PM was increased, as compared to macrophages treated with supernatant or sera from control rats not infected with T. lewisi. This effect was observed only if the active supernatant or sera came from rats infected with the T. lewisi 3 to 6 d before Toxoplasma inoculation. Furthermore, immunosuppressive activity was only detectable after at least 24 h incubation with the supernatant or sera. These results are in accordance with our in vivo previous studies which demonstrated a clear immunosuppressive effect of T. lewisi during T.gondii infection of the remarkably resistant Wistar rats