This study examined the relation of self-reported stress to cardiac autonomic modulation in real-life conditions. The participants for the study were healthy male (N = 59) and female (N = 40) employees (age 40 ± 10 years). A single-item question and a 14-item questionnaire on perceived stress were administered to the participants before the experimental night. RR-intervals (RRI) were recorded during night sleep and an orthostatic test after awakening at home. The RRI data were analyzed for heart rate (HR) and heart rate variability (HRV) in time and frequency domains. Nocturnal urinary stress hormone (cortisol, adrenal and noradrenal) secretion was also analyzed. Based on the self-reported stress, the participants were divided into either low or high stress group. The results showed that higher incidence of stress symptoms was significantly associated with lower HRV in the orthostatic test regardless of age and gender. Differences between the stress groups in HRV indices were approximately 20-50 and 30-75% in supine and standing positions, respectively. No difference was found in nocturnal HR, HRV, or stress hormone secretion between the stress groups. Higher incidence of stress symptoms was significantly associated with greater decrease of HRV from night sleep to the orthostatic test, as a response to awakening. In conclusion, the present findings support the view that autonomic modulation measured in the orthostatic test, but not during night sleep, is related to self-reported stress.