Stimulation of human peripheral blood lymphocytes by lipopolysaccharide (LPS) was studied by the incorporation of 3H-thymidine. Peak stimulation occurred at 7 to 9 days over a broad range of LPS concentrations. Both Escherichia coli and Salmonella typhimurium LPS were effective mitogens with S. typhimurium having slightly higher activity. There was a strict serum requirement; pooled fresh frozen human serum was found to best support stimulation. In fetal calf serum, LPS caused a reduction in culture-induced stimulation. Cell separation procedures were employed in order to study the nature of the responding cell population. It was found that only non-T cells were stimulated by LPS, but in order for maximal stimulation to occur there was a requirement for helper T cells.