A number of studies have now examined the association between smoking and the magnitude of physiological reactions to acute psychological stress. However, no large-scale study has demonstrated this association incorporating neuroendocrine in addition to cardiovascular reactions to stress. The present study compared neuroendocrine and cardiovascular reactions to acute stress exposure in current smokers, ex-smokers, and those who had never smoked in a large community sample. Salivary cortisol, systolic and diastolic blood pressure, heart rate and frequency components of systolic blood pressure and heart rate variability were measured at rest and during exposure to a battery of three standardized stress tasks in 480 male and female participants from the Dutch Famine Birth Cohort Study. Current smokers had significantly lower cortisol, systolic and diastolic blood pressure, and heart rate reactions to stress. They also exhibited smaller changes in the low frequency band of blood pressure variability compared to ex- and never smokers. There were no group differences in stress related changes in overall heart rate variability as measured by the root mean square of successive interbeat interval differences or in the high frequency band of heart rate variability. In all cases, effects remained significant following statistical adjustment for a host of variables likely to be associated with reactivity and/or smoking. In secondary analyses, there were no significant associations between lifetime cigarette consumption or current consumption and stress reactivity. In conclusion, compared to non-smokers and ex-smokers, current smokers exhibited attenuated neuroendocrine and cardiovascular reactions to acute psychological stress. Among smokers and ex-smokers, there is no evidence that lifetime exposure was associated with physiological reactions to acute stress, nor that current levels of cigarette consumption were associated with reactivity. It is possible, then, that attenuated stress reactivity may be a marker for an increased susceptibility to take up and/or maintain smoking behaviour once initiated.