To test the hypothesis that the rapid immune response of rats to Trypanosoma lewisi is elicited by prior exposure to cross-reacting environmental antigens, the early immune response to infection with this nonpathogenic protozoan was studied in germ-free and conventional rats. In germ-free rats, initial levels of both IgG and IgM were significantly lower than those of conventional rats. After infection, the germ-free rats made more immunoglobulins of both classes, and made them more quickly, than did conventional rats. Trypanosome-specific antibodies appeared earlier and in higher titers in the germ-free rats. Because they lacked intestinal microflora, it is unlikely that the germ-free rats' responses had been primed; thus, these observations indicated that the conventional rats' responses to some trypanosome antigens had been down-regulated by their prior exposure to environmental antigens. However, protective antibodies that inhibited parasite reproduction (ablastin) may have been primed, because these appeared in sera 2 days earlier in conventional rats. Despite much lower rates of production of trypanosome-specific antibodies, the conventional rats had the same peak parasitemias and times to crisis as germ-free rats. Thus it is apparent that protective immunity to this nonpathogenic parasite is not down-regulated by prior exposure to environmental antigens, as would be predicted.