Correction of metabolic deficiencies in the leukocytes of patients with chronic granulomatous disease
01 May 1970-Journal of Clinical Investigation (American Society for Clinical Investigation)-Vol. 49, Iss: 5, pp 865-870
TL;DR: Evidence is strengthened that the fundamental metabolic lesion in CGD cells during phagocytosis is indeed deficient production of hydrogen peroxide, probably, as previously shown, due to diminished oxidase for reduced nicotinamide adenine dinucleotide.
Abstract: Polymorphonuclear leukocytes from patients with chronic granulomatous disease (CGD) exhibit metabolic and bactericidal deficiencies that may be the result of inadequate production of H2O2. A hydrogen peroxide-generating system was, therefore, inserted into CGD leukocytes. This was accomplished by allowing the cells to phagocytize latex spherules coated with glucose oxidase. This produced an amelioration in the known metabolic deficiencies of these cells during phagocytosis: (a) intracellular (catalatic) formate oxidation dependent upon hydrogen peroxide production was enhanced fourfold; and (b) hexose monophosphate shunt activity, which other workers have shown to be at least partially dependent upon the availability of H2O2, was markedly stimulated. These data strengthen the evidence that the fundamental metabolic lesion in CGD cells during phagocytosis is indeed deficient production of hydrogen peroxide, probably, as previously shown, due to diminished oxidase for reduced nicotinamide adenine dinucleotide.
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1,710 citations
TL;DR: Results indicate that an O 2 − -generating system may be activated in potato tissues during the incompatible interaction induced by invading fungi or fungal wall components, and also that the generation of O 1 − may be involved during hypersensitive cell death as a trigger of the sequence of resistance reactions.
Abstract: Potato tuber tissue discs, which were aged after wounding in order to acquire hypersensitive reactivity, reduced extracellular cytochrome c and nitroblue tetrazolium (NBT) following inoculation with an incompatible, but not compatible, race of Phytophthora infestans . The cytochrome c -reducing activity rapidly increased from l to 4 h after inoculation along with an increase in the percentage of hypersensitively dead cells, and then decreased from the time when most of the penetrated cells had died. A localized activation of NBT reduction around invading hyphae of the incompatible, but not those of the compatible, race was observed at early stages of penetration before cell death. The reductive activity of the discs was also elicited by treatment with a hypersensitivity-eliciting substance, hyphal wall components of the fungus. Superoxide dismutase (SOD), an enzyme catalysing the conversion of the superoxide anion (O 2 − ) to H 2 O 2 and O 2 inhibited the enhanced reducing activity of the discs when added to the assay solution, indicating that cytochrome c and NBT may be reduced by O 2 − generated from the discs. Pre-infectional, vacuum infiltration of the discs with a solution containing SOD significantly delayed the occurrence of hypersensitive cell death caused by infection with the incompatible race as well as the accumulation of phytoalexin. Application of SH-binding reagents and NADP + , but not respiratory inhibitors, inhibited the elicitation of the reducing activity caused by infection with the incompatible race. These results indicate that an O 2 − -generating system may be activated in potato tissues during the incompatible interaction induced by invading fungi or fungal wall components, and also that the generation of O 2 − may be involved during hypersensitive cell death as a trigger of the sequence of resistance reactions.
773 citations
TL;DR: Particular organisms are susceptible to more than one antimicrobial system and thus may be effectively handled by back-up systems when one is absent, and an organism normally killed by the peroxidase system may be handled less efficiently but adequately when MPO is absent by other oxygen-dependent antimicrobial systems.
593 citations
TL;DR: Superoxide-dependent cytochrome c reduction was used to measure Superoxide production by granulocytes from two patients with chronic granulomatous disease and from children of similar age with similar age who had similar age-like symptoms.
Abstract: Superoxide-dependent cytochrome c reduction was used to measure Superoxide production by granulocytes from two patients with chronic granulomatous disease, from children of similar age wit...
348 citations
TL;DR: The azide-insensitive antimicrobial systems are more highly developed in peroxidase-negative leukocyte than in normal leukocytes, thus suggesting an adaptation.
Abstract: Azide and, to a lesser extent, cyanide inhibit the microbicidal activity of myeloperoxidase and of intact normal leukocytes, but they have little or no effect on peroxidase-negative leukocytes. The contribution of the azide-sensitive (peroxidase-dependent?) systems to the total microbicidal activity of normal leukocytes is considerable. The azide-insensitive antimicrobial systems are more highly developed in peroxidase-negative leukocytes than in normal leukocytes, thus suggesting an adaptation.
339 citations
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TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
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TL;DR: The deficiency of bactericidal capacity and the minimal degranulation after active phagocytosis by the PMN of these children with an inherited syndrome suggest that separate metabolic processes are involved in phagocytetosis and in intracellular digestion.
Abstract: Diminished bactericidal capacity was found to be characteristic of polymorphonuclear leukocytes (PMN) from five children with the clinical syndrome of granulomatous disease of childhood. The PMN from these children demonstrated nearly normal phagocytic capacity, and the majority of viable bacteria, after 2 hours of incubation in the phagocytosis system, were found associated with leukocytes. The morphology of the unstimulated polymorphonuclear leukocytes from patients with chronic granulomatous disease was similar to those from normal persons of similar ages by light and electron microscopy. In addition, the total lysozyme and phagocytin activity of leukocyte extracts from these patients was similar to those from equal numbers of leukocytes from controls.A striking difference in the cytoplasmic response after phagocytosis characterized the PMN of the patients with granulomatous disease. Whereas degranulation, vacuole formation, and rapid bacterial digestion were the rule in the PMN from controls, little degranulation and persistence of intact bacteria in the cytoplasm characterized disease. The deficiency of bactericidal capacity and the minimal degranulation after active phagocytosis by the PMN of these children with an inherited syndrome suggest that separate metabolic processes are involved in phagocytosis and in intracellular digestion. Continuing study of the metabolic function of leukocytes from these children should provide an opportunity for increased understanding of the metabolic basis for degranulation and intracellular digestion in phagocytic cells.
906 citations
TL;DR: Chronic granulomatous disease is an X-linked defect in the killing of certain bacteria by peripheral blood granulocytes and may be detected with the nitroblue tetrazolium (NBT) test.
Abstract: Chronic granulomatous disease is an X-linked defect in the killing of certain bacteria by peripheral blood granulocytes and may be detected with the nitroblue tetrazolium (NBT) test. The r...
860 citations
TL;DR: Myeloperoxidase, at high concentrations, exerted an antibacterial effect on L. acidophilus in the absence of added halide, which also was temperature- and catalase-sensitive, and suggests that, under the conditions employed, the antibacterial properties of a weak acid extract of guinea pig leukocytes is due, in part, to its peroxid enzyme content, particularly if a halide is present in the reaction mixture.
Abstract: An antibacterial effect of myeloperoxidase, a halide, such as iodide, bromide, or chloride ion, and H2O2 on Escherichia coli or Lactobacillus acidophilus is described. When L. acidophilus was employed, the addition of H2O2 was not required; however, the protective effect of catalase suggested that, in this instance, H2O2 was generated by the organisms. The antibacterial effect was largely prevented by preheating the myeloperoxidase at 80 C or greater for 10 min or by the addition of a number of inhibitors; it was most active at the most acid pH employed (5.0). Lactoperoxidase was considerably less effective than was myeloperoxidase when chloride was the halide employed. Myeloperoxidase, at high concentrations, exerted an antibacterial effect on L. acidophilus in the absence of added halide, which also was temperature- and catalase-sensitive. Peroxidase was extracted from intact guinea pig leukocytes by weak acid, and the extract with peroxidase activity had antibacterial properties which were similar, in many respects, to those of the purified preparation of myeloperoxidase. Under appropriate conditions, the antibacterial effect was increased by halides and by H2O2 and was decreased by catalase, as well as by cyanide, azide, Tapazole, and thiosulfate. This suggests that, under the conditions employed, the antibacterial properties of a weak acid extract of guinea pig leukocytes is due, in part, to its peroxidase content, particularly if a halide is present in the reaction mixture. A heat-stable antibacterial agent or agents also appear to be present in the extract.
819 citations