The characterization of superoxide production of human neonatal neutrophil

To assess the role of neutrophil in neonatal host defense against microbial infection, we characterized the superoxide anion (O(2-)) production of neonatal neutrophil on a biochemical basis. After taking an appropriate informed consent, neutrophils were obtained from cord blood immediately after transvaginal delivery and divided into two groups: the Preterm group, 15 neonates (27-36 weeks' gestation) and the Term group, 15 neonates (37-41 weeks' gestation). Eleven healthy adults served as controls in the Adult group. The value of N-formyl-methionyl-leucyl-phenylalanin (fMLP)-induced O(2-) production activity of neutrophils in the Preterm group using chemiluminescence assay was significantly lower than those values in both the Term and Adult groups (5.77+/-0.53x10(6) vs. 11.1+/-0.94x10(6) and 10.7+/-0.63x10(6) cpm; mean+/-S.E.M., p<0.05). In phorbol 12-myristate 13-acetate (PMA)-stimulation, the values of O(2-) production activity of neutrophils in both the Preterm and Term groups were significantly lower than that in the Adult group (13.0+/-1.66x10(6) and 18.0+/-1.44x10(6) vs. 27.3+/-1.45x10(6) cpm, p<0.05). Scatchard analysis of [(3)H]fMLP binding to neutrophil demonstrated a two-receptor model in each group, and the number of high-affinity receptors per neutrophil in the Preterm group was significantly lower than those in other groups (p<0.05). However, cord blood levels of proinflammatory cytokines, such as interleukin (IL)-6, -8, and tumor necrosis factor-alpha (TNF-alpha) did not differ in either neonatal group. These results indicated that the fMLP-induced O(2-)production activity of neutrophils in the term neonates was enhanced at the level of the receptor and suggested that this enhanced production contribute to the neonatal host defense against microbial infection.