Background Although the Trier Social Stress Test (TSST) constitutes a valid paradigm for social stress induction, less is known about the effects of a virtual reality (VR) TSST on short- and long-term hypothalamic-pituitary-adrenal (HPA) axis and sympathetic-adreno-medullar (SAM) axis responses. Hence, this study set out to evaluate reactivity and habituation of self-reported stress and HPA and SAM reactivity in a real TSST and VR-TSST when compared to a placebo TSST. Method Sixty-eight healthy young adults (50% female) were randomly assigned to either a real TSST, a VR-TSST, or a placebo TSST, all of which were conducted three times (one day and one week post initial exposure). Social presence, self-reported stress, salivary cortisol, heart rate (HR), and heart rate variability (HRV) were analyzed using ANOVAs and multilevel models. Findings On the first exposure, both the real and VR-TSST showed significantly stronger cortisol and cardiovascular responses than the placebo. On the second visit, the cortisol response was still significantly high—and the HRV response low—for the real and VR-TSST. The third visit resulted in HR, HRV, and cortisol responses comparable to the placebo group. Furthermore, the real TSST induced more self-reported stress than the placebo on all three visits, the VR-TSST only on the first two visits. Social presence was stable across conditions and had no association with stress markers. Conclusion These findings imply that the replicability of stress exposures at shorter intervals seems problematic for the traditional TSST, and for the VR-TSST.

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