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Transport workers face various occupational hazards, however long-term effects on mortality are less understood. Transport for London (TfL) employs almost 30,000 workers across a wide range of transport-based jobs and working environments. This study aimed to characterise mortality among TfL employees, and investigate long-term health outcomes. A retrospective cohort was formed using cause of death data from the TfL pension fund for employees working between 1960 and 2010. Workers were grouped by job title and Cox proportional hazard models were used to assess all-cause, respiratory, cardiovascular, and cancer mortality. Bus (hazard ratio HR 1.17, 95% confidence interval CI 1.09-1.25) and London Underground (LU) (HR 1.23, 95% CI 1.15-1.32) workers had significantly higher risks of all-cause, as well as respiratory, cardiovascular, and cancer mortality when compared to office workers. Mortality rates did not differ significantly between bus and LU workers, potentially due to shared occupational or lifestyle risk factors. In this large subway cohort study, mortality rates over 50 years were greater among bus and LU workers compared to office employees. However, findings should be interpreted cautiously due to limitations in data availability and unmeasured confounders. Future prospective studies should address these limitations by collecting detailed health and exposure data.