It has been suggested that infection by some mycoplasma species can act as possible cofactors in the acceleration of immunodeficiency in HIV-infected patients. The present study was designed to examine infections by oral mycoplasma species in HIV-seropositive (HIV(+)) patients. Mycoplasma salivarium and Mycoplasma orale were isolated from 59.5% and 16.7% of 42 HIV(+) patients, respectively. Non-M. salivarium and non-M. orale species were isolated from 40.5% of saliva samples from the HIV(+) group and 20.8% of those from 24 HIV-seronegative (HIV(-)) subjects, respectively. Although the production of superantigen by human peripheral lymphocytes in the isolated mycoplasma species from HIV(+) and HIV(-) subjects was evaluated, none of the examined mycoplasma strains, including ATCC strains of M. salivarium, M. orale, Mycoplasma buccae and Mycoplasma penetrans, were found to produce superantigen. Production of heat shock proteins (HSPs) by isolated mycoplasma strains was examined by immunoblotting using monoclonal antibodies against Helicobacter pylori HSP60. It was found that all the strains of M. salivarium, M. orale, and unidentified mycoplasma species isolated from HIV(+) and HIV(-) groups produced heat shock proteins. HSP production by oral mycoplasma may play a role in the immunomodulation of HIV(+) patients.