Separately, acute stress and cigarette cues are associated with cigarette smoking relapse, and each has been shown to increase nicotine seeking/self-administration. However, their potentially additive effects are understudied in humans. Using functional neuroimaging and a placebo-controlled double-blind design, we found acute stress suppressed cue-evoked activation in the medial orbitofrontal, parietal, and prefrontal cortices. Further, the effects of stress on nicotine withdrawal severity were inversely related to medial orbitofrontal and nucleus accumbens activation. Our findings illustrate acute stress exerts cooperative modulation of neural signals and subjective withdrawal severity, known to be important for long-term abstinence.