Nasopharyngeal colonization with Streptococcus pneumoniae precedes invasive pneumococcal disease. Human immunodeficiency virus (HIV) infection increases rates of invasive pneumococcal disease, and its effect on colonization is unknown. In a longitudinal cohort of Zambian mothers with or without HIV infection, HIV infection increased the risk of colonization (risk ratio [RR], 1.9; 95% confidence interval [CI], 1.3-2.8) and repeat colonization (RR, 2.4; 95% CI, 1.1-5.3) and reduced the time to new colonization (P = .01). Repeat colonization with homologous sero/factor types occurred only among HIV-positive mothers. Pediatric serotypes 6, 19, and 23 accounted for excess colonization among HIV-positive mothers. HIV infection significantly increases the risk of pneumococcal colonization. Increased rates of colonization by pediatric serotypes suggest a potential role for the 7-valent pneumococcal vaccine in HIV-infected adults.