Psychosocial stress elicited strong cortisol and neural responses in participants with burnout, with men showing higher cortisol reactivity, aligning with sex-specific cortisol stress response patterns. Stress activated limbic and task-positive regions but not the amygdala, possibly reflecting desensitization. While no general sex effects in neural activation were found, sex-specific associations emerged between cortisol and (para-) hippocampal responses. Resting-state connectivity indicated persistent limbic–task-positive coupling, underscoring shared and sex-dependent mechanisms of stress in burnout.