The virtual version of the Trier Social Stress Test (V-TSST) is an effective and standardized tool for social stress induction. This study aimed to examine gut permeability and physiological and inflammatory markers of reactivity to acute psychosocial stress. Forty young men were classified as high-stressed (HIGHS) or low-stressed (LOWS) according to the Shirom-Melamed Burnout Questionnaire. Cardiovascular reactivity and gut dysfunction were studied along with cortisol, zonulin and cytokines. Gut permeability was shown to be affected within one hour after the psychosocial stress induction, and shown to be dependent on age. Interleukin-6 increased with time, most pronounced at the end of the one-hour recovery after V-TSST, and was positively correlated to age. HIGHS experienced more abdominal dysfunction compared to LOWS. In conclusion, this study is the first to show fluctuations in gut permeability after psychosocial stress induction. This was partly associated with changes in inflammatory markers.