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Objectives: We investigated whether physical activity is associated with greater well-being in people with multiple long-term conditions or limiting long-term illness (LLI)., Study Design: Cross-sectional analysis of data from the Health Survey for England 2016., Methods: The Warwick-Edinburgh mental well-being score (WEMWBS) was evaluated according to number of days per week with >30 min moderate or vigorous activity. LLI and number of long-term conditions were evaluated as effect modifiers, adjusting for age, sex, smoking, body mass index and education. Marginal effects were estimated for female non-smokers, aged 45-54 years., Results: Data were analyzed for 5952 adults (female, 3275; male, 2677) including 1104 (19%) with non-limiting long-term illness and 1486 (25%) with LLI. There were 2065 (35%) with 1-2 long-term conditions, 461 (8%) with 3-4 and 58 (1%) with 5-6 long-term conditions. Participants with LLI were less likely to engage in physical activity on 5 or more days per week (LLI, 24%; No LLI, 47%) and more likely to be inactive (LLI, 41%; No LLI 13%). The adjusted marginal mean WEMWBS for inactive participants with no long-term illness was 49.0 (95% confidence interval 48.1 to 50.0), compared with 51.1 (50.4-51.8) if active on 5+ days per week. In LLI, the adjusted marginal mean WEMWBS was 41.6 (40.7-42.5) if inactive but 47.6 (46.6-48.6) if active on 5+ days per week. Similar associations were observed for the number of long-term conditions., Conclusions: Physical activity may be associated with greater increments in well-being among people with multiple long-term conditions or LLI than those without., (Copyright © 2020 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)