Stability and test-retest reliability of different hormonal stress markers upon exposure to psychosocial stress at a 4-month interval.

The Trier Social Stress Test (TSST) has been shown to reliably induce physiological stress responses in the hypothalamus-pituitary-adrenal (HPA) and in the sympathetic-adrenal-medullary (SAM) axis in cross-sectional studies. However, it was also reported that repeated exposure to the TSST might be associated with habituation, mainly of the HPA axis responsivity. Thus, in all longitudinal stress studies involving repeated TSST administration, potential habituation of the HPA axis response complicates the interpretation of results. The goal of the present study was therefore to assess stability and test-retest reliability of a number of different endocrinological stress markers as well as subjective stress responses after two exposures to the TSST four months apart. We assessed salivary and plasma cortisol profiles, plasma ACTH and noradrenaline profiles, as well as subjective stress ratings in healthy volunteers before, during, and after the TSST at six time-points both at test-day 1 (TSST_1, n = 42) and test-day 2 (TSST_2, n = 34) 4-months later. Half of the participants received the TSST in the early, the other half in the late afternoon. Discontinuous growth models were applied to model three phases of the stress response (preTSST, reactivity, recovery) for each marker. Subsequently, the stability of these phases was analyzed. Stability and test-retest reliability of standard physiological stress markers such as Area-under-the-Curve (AUCG, AUCI), Absolute Peak Change, and Relative Peak Change (RPC) were analyzed as well. We did not observe strong test-retest effects in any of the endocrinological measures. In contrast, test-retest effects in subjective stress were characterized by a faster drop directly after the second TSST, whereas the initial increase before the test period was the same for both test-days. Regarding test-retest-reliability, AUCG was the most reliable measure across all endocrinological and subjective stress markers (range: r = .606 to .858), while AUCI and RPC (range: r = - .146 to .548) were least reliable. A 4-month interval is a sufficient time interval between two repeated TSST exposures to largely reinstate the physiological stress response, which was also true for the initial psychological stress response. Thus, the TSST is well applicable in longitudinal studies. Keywords: Cortisol; Habituation; Hypothalamus-pituitary-adrenal axis reactivity; Subjective stress; Trier social stress test.