Reconstitution of bactericidal activity in chronic granulomatous disease cells by glucose-oxidase-containing liposomes.
Chronic granulomatous disease (CGD) is an inherited primary immunodeficiency characterized by phagocytes devoid of a functioning nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. The failure of CGD phagocytes to produce reactive oxygen species (ROS) results in a marked increase in the susceptibility of affected patients to life-threatening bacterial and fungal infections. This study investigated whether loading of CGD phagocytes with glucose oxidase (GO)-containing liposomes (GOLs) could restore cellular production of bactericidal ROS (eg, H2O2 and HOCl) in vitro. Results indicate that GO encapsulated in liposomes enabled NADPH oxidase-deficient phagocytes to use H2O2 for the production of highly bactericidal HOCl. The intracellular colocalization of bacteria and liposomes (or liposome-derived ferritin) was demonstrated by confocal laser microscopy and electron microscopy. After uptake of GOLs (approximately 0.2 U/mL at 1 mM total lipid concentration, size approximately 180 nm), CGD granulocytes produced HOCl levels comparable to those of normal phagocytes. Remarkably, after treatment with GOLs, CGD phagocytes killed Staphylococcus aureus as efficiently as normal granulocytes. Moreover, treated cells retained sufficient motility toward chemotactic stimuli as measured by chemotaxis assay. Side effects were evaluated by measuring the H2O2 concentrations and the production of methemoglobin in whole blood. These studies revealed that H2O2 produced by GOLs was degraded immediately by the antioxidative capacity of whole blood. Elevated methemoglobin levels were observed only after application of extremely high amounts of GOLs (2 U/mL). In summary, the application of negatively charged GOLs might provide a novel effective approach in the treatment of patients with CGD at high risk for life-threatening infections.
Gerber
C E
CE
University Children's Hospital, Department of Hematology and Oncology, Tübingen, Germany. claudia.gerber@med-tuebingen.de
Bruchelt
G
G
Falk
U B
UB
Kimpfler
A
A
Hauschild
O
O
Kuçi
S
S
Bächi
T
T
Niethammer
D
D
Schubert
R
R
D001769:Blood / Q000187:drug effects; D002478:Cells, Cultured; D002633:Chemotaxis; D004339:Drug Compounding; D005949:Glucose Oxidase / Q000008:administration & dosage / Q000494:pharmacology*; D006105:Granulomatous Disease, Chronic / Q000097:blood / Q000473:pathology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006997:Hypochlorous Acid / Q000378:metabolism; D008081:Liposomes; D008706:Methemoglobin / Q000096:biosynthesis; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D009504:Neutrophils / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D010587:Phagocytosis; D017382:Reactive Oxygen Species; D013211:Staphylococcus aureus
D008081:Liposomes; D017382:Reactive Oxygen Species; D006997:Hypochlorous Acid; D008706:Methemoglobin; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase