Group B streptococcus (GBS) is an important cause of early-and late-onset sepsis in the newborn. Preterm infants have markedly increased susceptibility and worse outcomes, but their immunological responses to GBS are poorly defined. We compared mononuclear cell and whole-blood cytokine responses to heat-killed GBS (HKGBS) of preterm infants (gestational age [GA], 26 to 33 weeks), term infants, and healthy adults. We investigated the kinetics and cell source of induced cytokines and quantified HKGBS phagocytosis. HKGBSinduced tumor necrosis factor (TNF) and interleukin 6 (IL-6) secretion was significantly impaired in preterm infants compared to that in term infants and adults. These cytokines were predominantly monocytic in origin, and production was intrinsically linked to HKGBS phagocytosis. Very preterm infants (GA, <30 weeks) had fewer cytokine-producing monocytes, but nonopsonic phagocytosis ability was comparable to that for term infants and adults. Exogenous complement supplementation increased phagocytosis in all groups, as well as the proportion of preterm monocytes producing IL-6, but for very preterm infants, responses were still deficient. Similar defective preterm monocyte responses were observed in fresh whole cord blood stimulated with live GBS. Lymphocyte-associated cytokines were significantly deficient for both preterm and term infants compared to levels for adults. These findings indicate that a subset of preterm monocytes do not respond to GBS, a defect compounded by generalized weaker lymphocyte responses in newborns. Together these deficient responses may increase the susceptibility of preterm infants to GBS infection.Streptococcus agalactiae (group B streptococcus [GBS]) is the leading infectious cause of death in the newborn (17), causing both early-and late-onset neonatal sepsis (EOS and LOS), characterized by septicemia, shock, and meningitis (16,18,31). The proportion of infants surviving extreme prematurity is increasing (11), and as a result preterm delivery is now the leading cause of neonatal morbidity and mortality. Preterm infants are exquisitely susceptible to both EOS and LOS; they suffer approximately a quarter of invasive GBS EOS cases and half of GBS LOS cases, with a Ͼ8-fold-greater mortality for EOS and 3-fold for LOS compared to term infants (29). While a proportion of the increased susceptibility to infection in preterm infants relates to the absence of passively acquired maternal IgG antibody, deficiencies in the early immune response to bacterial pathogens likely play a critical role (23). Several studies have addressed neonatal adaptive and humoral immune responses, such as complement activation and antibody production, but there has been little focus on innate immunity to bacterial pathogens and its relation to adaptive responses in this population (23,39).Innate immunity provides the first line of defense against bacterial infection and is therefore particularly important in the neonatal period (9). Additionally, the innate immune system, through activation of specif...