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The first human cases of monkeypox (MPX) were recognized in central Africa in 1970. Since then, sporadic outbreaks of the disease have occurred in central and west Africa. In 2003, an outbreak of human MPX occurred in the United States after importation of infected rodents from west Africa. Clinical features of the 2003 outbreak were less severe than accounts of the disease among people in central Africa. The reasons for this observed difference are unknown. In this study, the clinical and pathologic characteristics of experimental infection with representative central African and North American MPX virus strains were compared in a ground squirrel model of the disease. The results indicate that the US 2003 virus, which phylogenetically is a member of the west African MPX virus clade, was less virulent than central African MPX virus strains.