Little research has examined the impact of maternal lifetime trauma exposure on infant temperament. We examined associations between maternal trauma history and infant negative affectivity and modification by prenatal cortisol exposure in a sociodemographically diverse sample of mother–infant dyads. During pregnancy, mothers completed measures of lifetime trauma exposure and current stressors. Third-trimester cortisol output was assessed from maternal hair. When infants were 6 months old, mothers completed the Infant Behavior Questionnaire-Revised. In analyses that controlled for infant sex and maternal age, education, race/ethnicity, and stress during pregnancy, greater maternal trauma exposure was associated with increased infant distress to limitations and sadness. Higher and lower prenatal cortisol exposure modified the magnitude and direction of association between maternal trauma history and infant rate of recovery from arousal. The association between maternal trauma history and infant distress to limitations was somewhat stronger among infants exposed to higher levels of prenatal cortisol. The analyses suggested that maternal lifetime trauma exposure is associated with several domains of infant negative affectivity independently of maternal stress exposures during pregnancy and that some of these associations may be modified by prenatal cortisol exposure. The findings have implications for understanding the intergenerational impact of trauma exposure on child developmental outcomes.