Background: Variations in ankle injury rates and distributions among competition levels are unclear, but such data may help inform strategies to prevent ankle sprains during American football. Purpose: To describe the epidemiological patterns of ankle sprains in youth, high school (HS), and collegiate American football. Study Design: Descriptive epidemiological study. Methods: Data regarding youth, HS, and college football athletes were collected from 3 injury surveillance programs: (1) the Youth Football Safety Study (YFSS), (2) the National Athletic Treatment, Injury and Outcomes Network (NATION), and (3) the National Collegiate Athletic Association (NCAA) Injury Surveillance Program (ISP). During the 2012-2014 seasons, the YFSS, NATION, and NCAA ISP included 310, 184, and 71 football team-seasons, respectively. Athletic trainers (ATs) attended each practice and game and reported injuries and athlete-exposures (AEs) via their preferred injury documentation application. Ankle sprain rates for each type of ankle sprain were calculated overall, by event type (ie, practices and games), and specifically for severe injuries (ie, participation restriction time >21 days) and recurrent injuries (as defined by ATs). Rate ratios (RRs) were used to compare ankle sprain rates by competition level and event type. Injury proportion ratios (IPRs) were used to compare differences in severity, surgical needs, recurrence, injury mechanism, and injury activity by competition level. RRs and IPRs with 95% confidence intervals excluding 1.00 were considered statistically significant. Results: A total of 124, 897, and 643 ankle sprains were reported in youth, HS, and college football, respectively. This led to respective rates of 0.59, 0.73, and 1.19 sprains per 1000 AEs. The ankle sprain rate in college football was higher than the rates in HS (RR = 1.64; 95% CI, 1.48-1.82) and youth (RR = 2.01; 95% CI, 1.65-2.43) football. The proportion of ankle sprains that were recurrent in youth football was higher than the proportions in HS (IPR = 2.73; 95% CI, 1.68-4.50) and college (IPR = 2.19; 95% CI, 1.33-3.61) football. Conclusion: Ankle sprain rates were highest in college athletes. However, level-specific variations in ankle sprain severity and recurrence may highlight the need to develop level-specific policies and prevention strategies to reduce injury incidence.