Oxidase activity in cord blood neutrophils: A balance between increased membrane associated cytochrome b558 and deficient cytosolic components

Abstract

Newborn infants are prone to develop life-threatening pyogenic infections. Alterations in the function of neonatal phagocytes, including the activity of the neutrophil NADPH oxidase, have been suggested as one cause of increased susceptibility to such infections. In the present study, comprehensive analysis of NADPH oxidase enzyme system was performed in cord blood neutrophils from vaginally and cesarean section (CS) delivered, healthy, full-term infants. Superoxide anion (O(2) (-)) production by intact neutrophils from cord blood in response to soluble stimuli was equal to or increased compared to levels generated by cells from adult controls. In the sodium dodecyl sulfate (SDS) cell-free system, cytosol and plasma membrane from cord blood neutrophils generated O(2) (-) at comparable rates to subcellular fractions from healthy adults. However, mixing experiments demonstrated higher O(2) (-) generation with combination of cytosol from adult controls and membrane from cord blood neutrophils and lower O(2) (-) production with combination of cytosol from cord blood neutrophils and membrane from adult controls. Kinetic parameters for cord blood specimens were no different from those obtained for fractions from adult controls. Quantitative analysis of cytosolic components showed moderately reduced amount of p40-phox, p47-phox, and p67-phox in neutrophils from cord blood. In contrast, cytochrome b(558) content of plasma membrane of cord blood neutrophils was approximately 2-fold higher compared to adult controls. The normal to increased respiratory burst of intact cord blood neutrophils is the result of alterations to oxidase components: increased content of cytochrome b(558) in the plasma membrane and decreased levels of cytosolic components p47-phox, p67-phox, and p40-phox.