© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. Problem: To explore the impact of race and geographic region on biomarkers of HIV risk and vaginal health, differences in soluble immune mediators were measured in US versus African and US white versus US black women at enrollment into a phase 2 microbicide trial. Methods: Levels of soluble mucosal immune mediators and inhibitory activity against E. coli, which may serve as biomarkers of risk for HIV and other genital tract infections, were quantified in cervicovaginal lavage (CVL) collected from HIV-uninfected women in the United States (n = 73) and Africa (n = 73). Differences between groups were analyzed with multivariable logistic regression models for dichotomous variables and linear regression models for continuous variables. Results: Secretory leukocyte protease inhibitor, lactoferrin, human beta defensins, interleukin (IL)-8, and interferon-gamma-induced protein-10 were significantly higher in US compared to African women in multivariable analysis, but only IL-1β was significantly different between US white and black women. E. coli inhibitory activity did not differ among groups in adjusted analyses. Conclusion: Differences in soluble mucosal immunity between US and African women may play an important role in women's risk for HIV and other genital tract infections and response to prevention strategies including vaginal microbicides and should be considered in future studies.