Pseudomonas aeruginosa is a gram-negative bacillus that is a major cause of morbidity and mortality in immunosuppressed patients, burn patients, and patients with cystic fibrosis. Although immunity to these bacteria has been associated with serum antibody, more recent evidence suggests that T-cell-mediated immunity may also be important. To evaluate human T-cell responsiveness to these bacteria, the optimal conditions were determined for in vitro proliferation of human peripheral blood lymphocytes and T-lymphocytes to Fisher-Devlin immunotype 1 P. aeruginosa. The proliferative response of normal adult peripheral blood lymphocytes to heat-killed P. aeruginosa was studied in 34 subjects (range, 7,600 to 111,500 net cpm). Analysis of cell subpopulations indicated that T-lymphocytes are the major proliferating cells and that this response is enhanced by the presence of adherent cells. Data from fetal cord lymphocyte responses suggest that the proliferation seen in normal adult lymphocytes is induced by antigenic and not mitogenic stimulation.