The concentration of the iron-binding protein lactoferrin was measured in vaginal mucus of women throughout the menstrual cycle. Lactoferrin is proposed to limit growth of mucosal pathogens such as Neisseria gonorrhoeae. Lactoferrin concentration in mucus was highest just after menses (62.9 to 218 micrograms/mg of protein) and lowest (3.8 to 11.4 micrograms/mg of protein) just before menses. Overall concentration of mucus protein showed no variation with menstrual cycle, and little lactoferrin was detectable in cell debris. Plasma lactoferrin did not show the variation seen in vaginal mucus. Because of the suggestion of hormonal influence on vaginal lactoferrin, its concentration was determined in women receiving oral contraceptive therapy. Mean lactoferrin concentration in women taking oral contraceptives was significantly lower than in the control group (never exceeding 19.8 micrograms/mg of protein) and showed no monthly variation. Vaginal lactoferrin appears to be under hormonal control. Variation in vaginal lactoferrin concentration may result in alterations in susceptibility to bacterial pathogens such as Neisseriae gonorrhoeae.