scholarly journals Evaluation of Opsonic and Leukocyte Function With a Spectrophotometric Test in Patients With Infection and With Phagocytic Disorders

Blood ◽  
1973 ◽  
Vol 42 (1) ◽  
pp. 121-130 ◽  
Author(s):  
Thomas P. Stossel
Keyword(s):  
Bacterial Infections ◽  
Human Peripheral Blood ◽  
Normal Serum ◽  
Nitroblue Tetrazolium ◽  
Opsonic Activity ◽  
Nitroblue Tetrazolium Reduction ◽  
Leukocyte Function ◽  
Escherichia Coli Lipopolysaccharide ◽  
Tetrazolium Reduction ◽  
Oil Red O

Abstract Paraffin oil droplets containing Oil Red O and coated with Escherichia coli lipopolysaccharide were ingested rapidly by human peripheral blood phagocytes only if they were pretreated with normal serum. This property formed the basis of a screening test in which lipopolysaccharide-coated particles were opsonized with patients’ serums and then were added to autologous leukocytes suspended in two portions, one of which contained nitroblue tetrazolium. After these incubations the cells were washed and extracted with dioxane. Oil Red O and nitroblue tetrazolium formazan in the dioxane extracts were spectrophotometrically assayed, thereby providing simultaneous determinations of the initial rates of ingestion and nitroblue tetrazolium reduction. The test differentiated opsonically-deficient serums and chronic granulomatous disease phagocytes from normals. Serums from individuals with bacterial infections had supernormal opsonic activity, and leukocytes from these patients had increased rates of nitroblue tetrazolium reduction in response to ingestion.

scholarly journals Nitroblue Tetrazolium Reduction: False Positive and False Negative Results

Blood ◽  
1973 ◽  
Vol 41 (6) ◽  
pp. 921-925 ◽  
Author(s):  
Philip Ashburn ◽  
M. Robert Cooper ◽  
Charles E. McCall ◽  
Lawrence R. DeChatelet
Keyword(s):  
Bacterial Infection ◽  
Polycythemia Vera ◽  
False Positive ◽  
False Negative ◽  
Nitroblue Tetrazolium ◽  
Chronic Granulocytic Leukemia ◽  
Nitroblue Tetrazolium Reduction ◽  
Nbt Test ◽  
Granulocytic Leukemia ◽  
Tetrazolium Reduction

Abstract Quantitative and histochemical nitroblue tetrazolium reduction (NBT) tests were performed on leukocytes of 13 patients with polycythemia vera, seven with chronic granulocytic leukemia, eight with neoplastic disease associated with fever, 16 with bacterial infection, and 13 healthy control individuals. No significant differences were detected in the quantitative test between any of the groups studied. The histochemical NBT test was significantly higher than control in the patients with polycythemia vera and neoplasia associated with fever, as well as in those with known bacterial infection. It is suggested that patients with neoplasia or polycythemia vera may show a false positive reaction in the histochemical NBT test. Conversely, the test was significantly lower than control in patients with chronic granulocytic leukemia. Such patients might show a false negative reaction in the test even if infection were present.

scholarly journals Effect of dihydroheptaprenol on nitroblue tetrazolium reduction by swine alveolar macrophages.

1989 ◽  
Vol 51 (3) ◽  
pp. 630-631
Author(s):  
Toshihiro WATARI ◽  
Ryo GOITSUKA ◽  
Hidekazu KOYAMA ◽  
Toshinori SAKO ◽  
Tomiya UCHINO ◽  
...  
Keyword(s):  
Alveolar Macrophages ◽  
Nitroblue Tetrazolium ◽  
Nitroblue Tetrazolium Reduction ◽  
Tetrazolium Reduction

scholarly journals 1,25-Dihydroxyvitamin D3-induced differentiation in a human promyelocytic leukemia cell line (HL-60): receptor-mediated maturation to macrophage-like cells.

1984 ◽  
Vol 98 (2) ◽  
pp. 391-398 ◽  
Author(s):  
D J Mangelsdorf ◽  
H P Koeffler ◽  
C A Donaldson ◽  
J W Pike ◽  
M R Haussler
Keyword(s):  
Cell Line ◽  
Leukemia Cell ◽  
Promyelocytic Leukemia ◽  
Nitroblue Tetrazolium ◽  
Leukemia Cell Line ◽  
Induced Differentiation ◽  
Dihydroxyvitamin D3 ◽  
Nitroblue Tetrazolium Reduction ◽  
Tetrazolium Reduction ◽  
Promyelocytic Leukemia Cell Line

The human-derived promyelocytic leukemia cell line, HL-60, is known to differentiate into mature myeloid cells in the presence of 1,25-dihydroxyvitamin D3 (1,25[OH]2D3). We investigated differentiation by monitoring 1,25(OH)2D3-exposed HL-60 cells for phagocytic activity, ability to reduce nitroblue tetrazolium, binding of the chemotaxin N-formyl-methionyl-leucyl-[3H]phenylalanine, development of nonspecific acid esterase activity, and morphological maturation of Wright-Giemsa-stained cells. 1,25(OH)2D3 concentrations as low as 10(-10) M caused significant development of phagocytosis, nitroblue tetrazolium reduction, and the emergence of differentiated myeloid cells that had morphological characteristics of both metamyelocytes and monocytes. These cells were conclusively identified as monocytes/macrophages based upon their adherence to the plastic flasks and their content of the macrophage-characteristic nonspecific acid esterase enzyme. The estimated ED50 for 1,25(OH)2D3-induced differentiation based upon nitroblue tetrazolium reduction and N-formyl-methionyl-leucyl-[3H]phenylalanine binding was 5.7 X 10(-9) M. HL-60 cells exhibited a complex growth response with various levels of 1,25(OH)2D3: less than or equal to 10(-10) M had no detectable effect, 10(-9) M stimulated growth, and greater than or equal to 10(-8) M sharply inhibited proliferation. We also detected and quantitated the specific receptor for 1,25(OH)2D3 in HL-60 and HL-60 Blast, a sub-clone resistant to the growth and differentiation effects of 1,25(OH)2D3. The receptor in both lines was characterized as a DNA-binding protein that migrated at 3.3S on high-salt sucrose gradients. Unequivocal identification was provided by selective dissociation of the 1,25(OH)2D3-receptor complex with the mercurial reagent, p-chloromercuribenzenesulfonic acid, and by a shift in its sedimentation position upon complexing with anti-receptor monoclonal antibody. On the basis of labeling of whole cells with 1,25(OH)2[3H]D3 in culture, we found that HL-60 contains approximately 4,000 1,25(OH)2D3 receptor molecules per cell, while the nonresponsive HL-60 Blast is endowed with approximately 8% of that number. The concentration of 1,25(OH)2D3 (5 X 10(-9) M) in complete culture medium, which facilitates the saturation of receptors in HL-60 cells, is virtually identical to the ED50 for the sterol's induction of differentiation. This correspondence, plus the resistance of the relatively receptor-poor HL-60 Blast, indicates that 1,25(OH)2D3-induced differentiation of HL-60 cells to monocytes/macrophages is occurring via receptor-mediated events.

scholarly journals Combinations of recombinant human interferons and retinoic acid synergistically induce differentiation of the human promyelocytic leukemia cell line HL-60

Blood ◽  
1987 ◽  
Vol 69 (2) ◽  
pp. 501-507
Author(s):  
H Hemmi ◽  
TR Breitman
Keyword(s):  
Retinoic Acid ◽  
Cell Line ◽  
Leukemia Cell ◽  
Promyelocytic Leukemia ◽  
Nitroblue Tetrazolium ◽  
Leukemia Cell Line ◽  
Nonspecific Esterase ◽  
Nitroblue Tetrazolium Reduction ◽  
Tetrazolium Reduction ◽  
Promyelocytic Leukemia Cell Line

The human acute promyelocytic leukemia cell line HL-60 is induced to differentiate into morphologically and functionally mature monocytelike cells by incubation with a combination of 10 nmol/L retinoic acid (RA) and various concentrations of recombinant immune interferon (rIFN- gamma). These induced cells show marked increases in antibody-dependent cellular cytotoxicity (ADCC), antibody-coated erythrocyte (EA) rosettes, nonspecific esterase, and 5′-nucleotidase activity. rIFN- gamma alone at concentrations of 10 to 1,000 U/mL has essentially no effect on morphological maturation, nitroblue tetrazolium reduction, and immunophagocytosis. However, rIFN-gamma at these concentrations increases EA rosetting in a concentration-dependent manner that is not affected by 10 nmol/L RA. At a concentration of 1,000 U/ml, rIFN-gamma induces moderate increases in nonspecific esterase, 5′-nucleotidase, and ADCC. These parameters are markedly increased by the addition of 10 nM RA, a concentration which alone has no effect on these markers. Based on units of antiviral activity, rIFN-gamma is tenfold more active than rIFN-alpha D in inducing EA rosettes and 40-fold more active in inducing nitroblue tetrazolium reduction and immunophagocytosis. These results, indicating that combinations of rIFN-gamma or rIFN-alpha and RA synergistically induce differentiation of HL-60, suggest that this combination may have clinical utility in the treatment of patients with certain leukemias.

scholarly journals The biochemical basis of nitroblue tetrazolium reduction in normal human and chronic granulomatous disease polymorphonuclear leukocytes

Blood ◽  
1976 ◽  
Vol 48 (2) ◽  
pp. 309-313 ◽  
Author(s):  
RL Baehner ◽  
LA Boxer ◽  
J Davis
Keyword(s):  
Chronic Granulomatous Disease ◽  
Polymorphonuclear Leukocytes ◽  
Nitroblue Tetrazolium ◽  
Granulomatous Disease ◽  
Biochemical Basis ◽  
Nitroblue Tetrazolium Reduction ◽  
Normal Human ◽  
Nbt Reduction ◽  
Human Polymorphonuclear Leukocytes ◽  
Tetrazolium Reduction

Normal human polymorphonuclear leukocytes (PMN) placed in anaerobic chambers reaching pO2's of less than 5 mm Hg fail to generate O2-, iodinate ingested particles, and stimulate glucose-1–14C oxidation through the hexose monophosphate shunt. The observation that anaerobic cells are incapable of generating O2- or reducing nitroblue tetrazolium (NBT) to formazan supports the idea that NBT reduction in phagocytizing PMN is due exclusively to oxygen-dependent O2- generating oxidase which is deficient in chronic granulomatous disease leukocytes, despite their hyperphagocytic capacity.

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