Recent studies suggest that the chemokine eotaxin may participate in atherosclerosis. Threonine (T) for alanine (A) substitution at amino acid 23 in the eotaxin gene (CCL11) has been associated with risk of developing allergic-inflammatory disorders. However, no genetic-epidemiological data are available on the risk of cardiovascular disease associated with this polymorphism. Using DNA samples collected at baseline in a prospective cohort of 14,916 initially healthy American men, we evaluated the A23T polymorphism among 523 individuals who subsequently developed myocardial infarction (MI) and among 2092 individuals who remained free of reported cardiovascular disease over a mean follow-up period of 13.2 years. The T23 allele was significantly associated with risk of myocardial infarction (odds ratio (OR) in an age and smoking adjusted recessive model of inheritance, 1.86; 95% confidence interval (CI), 1.15-3.01; P = 0.012). This risk effect remained statistically significant in analyses further controlling for body mass index, history of hypertension, the presence of diabetes, and randomized treatment assignment (OR, 1.95; 95% CI, 1.19-3.18; P = 0.008). In this cohort, a T for A substitution at amino acid 23 in the eotaxin gene is associated with increased risk for incident myocardial infarction. If confirmed in other cohorts, these data support the emerging hypothesis that eotaxin participates in atherosclerosis.