This study addressed whether lifetime stressor exposure was associated with psychophysiological reactivity and habituation to a novel laboratory-based stressor. Eighty-six participants (Mage = 23.31 years, SD = 4.94) reported their exposure to lifetime non-sport and sport-specific stressors before completing two consecutive trials of the Trier Social Stress Test, while cardiovascular (i.e., heart rate) and endocrine (i.e., salivary cortisol) data were recorded. Exposure to a moderate number of lifetime non-sport and sport-specific stressors was associated with adaptive cardiovascular reactivity, whereas very low or very high stressor exposure was related to maladaptive reactivity. Moreover, experiencing a very low number of lifetime non-sport (but not sport-specific) stressors was associated with poorer habituation. In contrast, lifetime stressor severity was unrelated to cardiovascular reactivity. Finally, greater lifetime non-sport and sport-specific stressor counts were associated with blunted cortisol reactivity and poorer habituation. These results suggest that lifetime stressor exposure may influence sport performers’ acute stress responses.