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Work-related stress appears to be especially high among teachers. However, most research on teacher stress relies exclusively on teachers’ self-reports. Little is known about the physiological correlates of affective stress in teachers. This longitudinal study investigates the relationship between core self-evaluation and adverse psychological and physiological stress outcomes in 42 teachers. Self-report questionnaires were used to assess core self-evaluation, vital exhaustion, and resignation tendency. The concentration of cortisol was assessed using hair samples. One year after the initial measurement, vital exhaustion and hair cortisol were assessed again. Path-analytic mediational models showed that core self-evaluation strongly predicted vital exhaustion, and resignation tendency partially mediated this relationship. However, core self-evaluation did not predict hair cortisol concentration. These findings suggest that core self-evaluation plays a crucial role in preventing vital exhaustion among teachers. A positive core self-evaluation seems beneficial for teachers’ primary and secondary appraisal and an essential resource for the long-term prevention of self-reported vital exhaustion.