Protective effects of vaccination of hamsters against Mycoplasma pneumoniae infection, evaluated according to the recovery of mycoplasmas and histopathological changes in the respiratory tract after challenge infection, persisted for at least 6 months after the final vaccination. Serum antibody levels reached a maximum in the second week after the last vaccination and decreased markedly between the first and the third months, but increased again in sera obtained from animals given booster injections. Metabolism-inhibiting antibodies were detected in bronchial washings of animals showing high resistance obtained by vaccinal or passive immunization. Antiserum transfer was also effective for protection but cell-mediated immune responses were not demonstrated in any animals up to 6 months after the vaccination. Even after 10 months, suppression of both mycoplasmal proliferation and lung lesions was apparent, and a single dose of the vaccine induced a significant booster effect. These findings suggest that (1) humoral immunity is more important than cell-mediated immunity in resistance of hamsters to M. pneumoniae pneumonia, and (2) the antibody secreted in the respiratory tract may be involved in the local defense mechanisms.