Although 40% of US women indicate they are currently trying to lose weight, the association between intentional weight loss and longevity is unknown. The authors analyzed prospective data from 43,457 overweight, never-smoking US white women aged 40-64 years who in 1959-1960 completed a questionnaire that included questions on weight change direction, amount, time interval, and intentionality. Vital status was determined in 1972. Proportional hazards regression was used to estimate mortality rate ratios for women who intentionally lost weight compared with women who had no change in weight. Women who died within the first 3 years of follow-up were excluded. Analyses were stratified by preexisting illness and adjusted for age, beginning body mass index, alcohol intake, education, physical activity, and health conditions. In women with obesity-related health conditions (n = 15,069), intentional weight loss of any amount was associated with a 20% reduction in all-cause mortality, primarily due to a 40-50% reduction in mortality from obesity-related cancers; diabetes-associated mortality was also reduced by 30-40% in those who intentionally lost weight. In women with no preexisting illness (n = 28,388), intentional weight loss of > or = 20 lb (> or = 9.1 kg) that occurred within the previous year was associated with about a 25% reduction in all-cause, cardiovascular, and cancer mortality; however, loss of < 20 lb (< 9.1 kg) or loss that occurred over an interval of > or = 1 year was generally associated with small to modest increases in mortality. The association between intentional weight loss and longevity in middle-aged overweight women appears to depend on their health status. Intentional weight loss among women with obesity-related conditions is generally associated with decreased premature mortality, whereas among women with no preexisting illness, the association is equivocal.