BACKGROUND: The experience of social exclusion represents an extremely aversive and threatening situation in daily life. The present study examined the impact of social exclusion compared to inclusion on steroid hormone concentrations as well as on subjective affect ratings. METHODS: Eighty subjects (40 females) participated in two independent behavioral experiments. They engaged in a computerized ball tossing game in which they ostensibly played with two other players who deliberately excluded or included them, respectively. Hormone samples as well as mood ratings were taken before and after the game. RESULTS: Social exclusion led to a decrease in positive mood ratings and increased anger ratings. In contrast, social inclusion did not affect positive mood ratings, but decreased sadness ratings. Both conditions did not affect cortisol levels. Testosterone significantly decreased after being excluded in both genders, and increased after inclusion, but only in males. Interestingly, progesterone showed an increase after both conditions only in females. DISCUSSION: Our results suggest that social exclusion does not trigger a classical stress response but gender-specific changes in sex hormone levels. The testosterone decrease after being excluded in both genders, as well as the increase after inclusion in males can be interpreted within the framework of the biosocial status hypothesis. The progesterone increase might reflect a generalized affiliative response during social interaction in females.