Disseminated coccidioidomycosis is a life-threatening infection. In these studies, we examined protection against systemic murine coccidioidomycosis by vaccination with heat-killed Saccharomyces cerevisiae (HKY). CD-1 mice received HKY subcutaneously or by oral gavage with or without adjuvants once weekly beginning 3 or 4 weeks prior to infection; oral live Saccharomyces was also studied. All HKY sc regimens were equivalent, prolonging survival (P<or=0.005) and reducing fungal burden versus controls. Oral live Saccharomyces, but not HKY, prolonged survival (P=0.03), but did not reduce fungal burden. Survival of mice given HKY was equivalent to vaccination with formalin-killed spherules, but inferior in reduction of fungal burden. HKY was superior to a successful recombinant vaccine, PRA plus adjuvant. This novel heterologous protection afforded by HKY vaccination offers a new approach to a vaccine against coccidioidomycosis.