To characterize the human immune response to syphilis, we determined the effect of infection with Treponema pallidum on the percentage of the various lymphocyte subpopulations in the peripheral blood of infected and uninfected persons. Monoclonal antibodies labeled with either fluorescein isothiocyanate or phycoerythrin were used to perform dual color analysis on a FACScan with the following markers: CD3 for total T cells, CD4 for T helper cells, CD8 for T suppressor cells, CD19 for B cells, and CD16 plus CD56 for natural killer cells. Lymphocyte immunophenotype results were analyzed by the stage of untreated syphilis and by gender. Although they were within the ranges of the normal distribution of immunophenotypes, the percentages of CD4+ cells were significantly lower (P < 0.001) and those of CD8+ cells were higher (P = 0.03) in patients with syphilis than in the uninfected population. For infected versus uninfected subjects, both women and men, the differences in the mean percentages of CD3+ and CD4+ cells were significant (P < or = 0.05). Significant differences were noted between the sexes in secondary syphilis only in the mean percentages of cells positive for CD3, CD4, CD8, and CD16 plus CD56. Gender had no effect on lymphocyte subpopulations in subjects with primary or latent syphilis. In the control population, significant differences due to gender were observed in the percentages of cells positive for CD3, CD4, and CD16 plus CD56.