Due to elevated ozone concentrations at high altitudes, the adverse effect of ozone on air quality, human perception and health may be more pronounced in aircraft cabins. The association between ozone and passenger-reported symptoms has not been investigated under real conditions since smoking was banned on aircraft and ozone converters became more common. Indoor environmental parameters were measured at cruising altitude on 83 US domestic and international flights. Passengers completed a questionnaire about symptoms and satisfaction with the indoor air quality. Average ozone concentrations were relatively low (median: 9.5 ppb). On thirteen flights (16%) ozone levels exceeded 60 ppb, while the highest peak level reached 256 ppb for a single flight. The most commonly reported symptoms were dry mouth or lips (26%), dry eyes (22.1%) and nasal stuffiness (18.9%). 46% of passengers reported at least one symptom related to the eyes or mouth. A third of the passengers reported at least one upper respiratory symptom. Using multivariate logistic (individual symptoms) and linear (aggregated continuous symptom variables) regression, ozone was consistently associated with symptoms related to the eyes and certain upper respiratory endpoints. A concentration-response relationship was observed for nasal stuffiness and eye and upper respiratory symptom indicators. Average ozone levels, as opposed to peak concentrations, exhibited slightly weaker associations. Medium and long duration flights were significantly associated with more symptoms compared to short flights. The relationship between ultrafine particles and ozone on flights without meal service was indicative of ozone-initiated chemistry.