Description of the pathobiology of the recently described zoonotic agent of human ehrlichiosis (Ehrlichia chaffeensis) would be greatly facilitated by the availability of a convenient experimental animal model of infection. We determined whether C3H/HeJ mice could sustain persistent infection by this predominantly monocyte-inhabiting rickettsia. Such mice rapidly produced an intense specific IgG response upon inoculation of ehrlichiae, and high titers were demonstrable for more than 6 months thereafter. Ehrlichiae were reisolated from the peripheral blood and spleen of 1 mouse at day 11 after inoculation. DNA of E. chaffeensis was more frequently detected within these tissues by polymerase chain reaction. Other candidate rodent models appeared to be poor hosts for this pathogen. About half of intact and virtually all splenectomized white-footed mice that were inoculated seroconverted. Sera from inoculated voles and hamsters did not react to antigens of E. chaffeensis. The C3H/Hej mouse becomes persistently infected by this rickettsia, and may serve as a useful model for studies of the immune response to the agent of human ehrlichiosis.