Nitric oxide (NO) has beneficial effects on cardiovascular and immune health. Stress and depression have been linked to a reduction in serum NO. In this study, we examined the effect of academic exam stress on the fraction of NO in exhaled air (FeNO) and spirometric lung function in 41 healthy college students. Participants completed assessments at mid-semester as well as in the early and late phase of an academic exam period. Negative affect, depressive mood, and salivary cortisol were elevated during exams, whereas FeNO and lung function decreased. Higher depressive mood was associated with lower FeNO, whereas higher negative affect was associated higher FeNO across time. These findings provide initial evidence that depression and prolonged stress can alter FeNO and lung function in healthy individuals, which could have adverse consequences for cardiovascular, airway, and immune health.