PEGylated D-amino acid oxidase restores bactericidal activity of neutrophils in chronic granulomatous disease via hypochlorite.
Chronic granulomatous disease (CGD) causes impaired hydrogen peroxide (H(2)O(2)) generation. Consequently, neutrophils in patients with CGD fail to kill infecting pathogens. We expected that supplementation with H(2)O(2) would effectively restore the bactericidal function of neutrophils in CGD. Here, we used polyethylene glycol-conjugated D-amino acid oxidase (PEG-DAO) as an H(2)O(2) source. The enzyme DAO generates H(2)O(2) by using D-amino acid and oxygen as substrates. PEG-DAO plus D-amino acid indeed exerted bacteriostatic activity against Staphylococcus aureus via H(2)O(2) in vitro. Furthermore, use of PEG-DAO plus D-amino acids, which increased the amount of intracellular H(2)O(2), restored bactericidal activity of neutrophils treated with diphenylene iodonium, in which nicotinamide adenine dinucleotide phosphate (NADPH) oxidase was defective. This restoration of bactericidal activity was mediated by myeloperoxidase, with concomitant production of H(2)O(2) by PEG-DAO plus D-Ala. We also confirmed that PEG-DAO treatment restored bactericidal activity of congenitally defective neutrophils from patients with CGD. These results indicate that PEG-DAO can supply additional H(2)O(2) for defective NADPH oxidase of neutrophils from patients with CGD, and thus neutrophils regain bactericidal activity.
Laboratory of Microbiology and Oncology, Faculty of Pharmaceutical Science, Sojo University, Ikeda 4-22-1, Kumamoto 860-0082, Japan.
Hideaki
H
Nakamura
Jun
J
Fang
Tomoyuki
T
Mizukami
Hiroyuki
H
Nunoi
Hiroshi
H
Maeda
D000818:Animals; D001770:Blood Bactericidal Activity / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D003605:D-Amino-Acid Oxidase / Q000494:pharmacology*; D005260:Female; D006105:Granulomatous Disease, Chronic / Q000378:metabolism / Q000503:physiopathology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006997:Hypochlorous Acid / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D008813:Mice, Inbred ICR; D023421:Models, Animal; D019255:NADPH Oxidases / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D009195:Peroxidase / Q000378:metabolism; D013211:Staphylococcus aureus / Q000502:physiology
D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide; D009195:Peroxidase; D003605:D-Amino-Acid Oxidase; D019255:NADPH Oxidases