High altitude (HA) is a multi-stressor environment comprising hypobaric hypoxia and cold. Climatic temperature varies with seasonal variation at HA. The present study was undertaken to investigate the effect of ambient temperature on antioxidant profile among sojourners at HA. The study was conducted on sojourners exposed to an altitude of 4,560 m in two different seasons and categorized into two groups (SOJ 1, n=63, ambient temp. at HA: -6 degree to +10degreeC; SOJ 2, n=81, ambient temp. at HA: 3degree-22degreeC). Blood was collected at sea level (SL) and after 4 weeks of HA exposure. Antioxidant enzymes showed significant upregulation in SOJ 2 at HA. In SOJ 1, superoxide dismutase and glutathione peroxidase showed significant upregulation but catalase and glutathione reductase showed significant decrease at HA. Non-enzymatic antioxidants showed significant reduction in SOJ 1 whereas a sustained antioxidant profile was observed in SOJ 2 at HA. Oxidative stress markers showed higher levels in SOJ 1 than SOJ 2 at HA. Differences observed between SOJ 1 and SOJ 2 at HA may be the consequence of different environmental temperatures. Cold stress was higher in SOJ 1 as evidenced from the significantly lower oral temperature in SOJ 1 as compared to SOJ 2. Cold- and hypoxia-induced increase in energy expenditure was significantly high in SOJ 1 than SOJ 2. To conclude, chronic exposure to hypoxia in moderate climatic temperature has a potential preconditioning effect on antioxidant system, but exposure to both cold and hypoxia causes greater oxidative stress due to altered metabolic rate.