This study examined the long-term effects of prematurity and perinatal adversity on individual differences in stress-related reactivity and regulation of the HPA axis. A prospective sample of 155 infants born preterm and healthy (n = 20), medical illness (n = 48), neurological illness (n = 26), and small for gestational age (n = 24) and full-term (n = 37) were recruited between 1985 and 1989. At age 23 years, multiple saliva samples were collected before and after participation in the Trier Social Stress Test and later assayed for cortisol. Results reveal that at age 23 years, infants born premature with neurological complications showed higher cortisol reactivity to social evaluative threat compared to either their full-term, small for gestation age, medically ill, or healthy preterm peers. Findings are discussed in terms of implications for contemporary theories that propose effects of early adversity on biological sensitivities and susceptibilities, which translate experience into developmental outcomes related to poor health and risk for disease.