DNA immunization offers a novel approach to inducing humoral and cellular immunity against infectious pathogens. We examined whether such an approach could be used against cryptosporiodiosis, an intestinal disease caused by the protozoan parasite Cryptosporidium parvum. This infection is a major problem for young ruminants and immunosuppressed individuals in whom cryptosporidiosis causes life-threatening symptoms. The life cycle of C. parvum takes place in the enterocytes of the intestinal epithelium. We therefore focused our attention on a route of immunization that might induce a mucosal immunoglobulin (Ig)A response. Eight-week-old BALB/c mice were immunized intranasally with DNA encoding a 15-kDa C. parvum sporozoite antigen (CP15-DNA) cloned onto the plasmid pcDNA3. CP15-DNA-immunized mice developed specific and longlasting production of anti-CP15 Ig A in intestinal secretions and specific IgG in sera 3 months and 1 year after the first DNA inoculation. CP15-DNA-immunized mice also developed an antigen-specific T lymphocyte proliferative response in both spleen and mesenteric lymph nodes. Control mice that received the pcDNA3 plasmid alone did not develop specific humoral and cellular responses. These results indicate that plasmid DNA may provide a powerful means of eliciting intestinal humoral and cellular responses to C. parvum infections in mammals.