A new formalin-inactivated vaccine prepared by sucrose density gradient centrifugation of tissue culture-grown Rickettsia rickettsii was evaluated for safety and immunogenicity in a placebo-controlled, double-blind study. Most of the 52 seronegative adult vaccinees, 88% after the first and 66% after the second dose, experienced brief, mild (mostly local) reactions, but only 50% exhibited a systemic immune response to vaccination. Six unvaccinated volunteers (controls) and 16 of these vaccinees were challenged with R rickettsii one month after vaccination. Vaccine efficacy was 25%; all six controls and 12 of 16 vaccinees developed typical Rocky Mountain spotted fever. The incubation period was longer, the duration of constitutional symptoms shorter, and the height of fever lower in ill vaccinees than in controls. The vaccine provided only partial protection against Rocky Mountain spotted fever but ameliorated the illness.