The purpose of this study was to disentangle the psychobiological mechanisms and social-evaluative conditions that mediate the process by which the Autonomic Nervous System reacts in male and female humans. We used the original Trier Social Stress Test, as well as two modifications to this original social stressor: a punishment modification and a reward modification. We obtained measures of autonomic (heart rate and respiratory sinus arrhythmia; HR and SA respectively) reactivity before, during and after the stress test. To distinguish the contribution of the different modifications and any additional difference in reactivity due to gender, the participants were randomly separated into the three modifications, where N=35 (17 male) for the no modification group, N=12 (7 male) for the punishment condition, and N=13 (8 male) for the reward condition. All participants exhibited ANS reactivity to the stressor; females exhibited the most magnified response to all modifications. Overall, the most ANS reactivity was found within the reward condition, with the no modification group exhibiting the least amount of reactivity. This suggests that the reward paradigm was the most salient of all the stressors. Evidence indicated that the ANS stress response system is highly sensitive to potential for gain and reward, especially in females.