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. 1974 Nov;54(5):1227–1234. doi: 10.1172/JCI107866

Enhanced Phagocytic Capacity THE BIOLOGIC BASIS FOR THE ELEVATED HISTOCHEMICAL NITROBLUE TETRAZOLIUM REACTION

Charles E McCall 1,2,3, Lawrence R DeChatelet 1,2,3, Robert Butler 1,2,3, David Brown 1,2,3
PMCID: PMC301670  PMID: 4153897

Abstract

The biologic basis for the elevated histochemical reduction of nitroblue tetrazolium dye (NBT) in neutrophils from patients with acute bacterial infection or polycythemia vera was studied. A precipitin reaction followed mixing NBT with heparin. NBT was reduced after phagocytosis of this complex (H-NBT) by polymorphonuclear leukocytes (PMNs). Ingestion required divalent cations and was facilitated by the presence of complement. H-NBT incubated with normal but not with C2-deficient human serum converted native C3 to its inactive form.

Phagocytic indices were determined in patients and controls by measuring O2 utilization and hexose monophosphate shunt activity and by visually counting cell-associated latex particles. Significant elevations above controls were observed in phagocytes isolated from all patients with elevated histochemical NBT scores when H-NBT complex, latex, or zymosan was employed as the phagocytic particle. Increased indices were observed in the presence of fresh AB serum, heat-inactivated AB serum, or without serum. Serum from patients with elevated NBT scores did not alter phagocytosis in control phagocytes. With NADH and NADPH as substrates, total NBT diaphorase activity of sonicated leukocytes was normal in all patients.

These results suggest that increased phagocytic capacity of PMNs is the primary cause of increased histochemical NBT reduction. The PMNs of patients with acute bacterial infection or polycythemia vera may have alterations in their cell membranes which lead to an enhanced rate of phagocytosis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashburn P., Cooper M. R., McCall C. E., DeChatelet L. R. Nitroblue tetrazolium reduction. False positive and false negative results. Blood. 1973 Jun;41(6):921–925. [PubMed] [Google Scholar]
  2. Brown D. L., Lachmann P. J. The behaviour of complement and platelets in lethal endotoxin shock in rabbits. Int Arch Allergy Appl Immunol. 1973;45(1):193–205. doi: 10.1159/000231028. [DOI] [PubMed] [Google Scholar]
  3. Cooper M. R., DeChatelet L. R., McCall C. E., Spurr C. L. The activated phagocyte of polycythemia vera. Blood. 1972 Sep;40(3):366–374. [PubMed] [Google Scholar]
  4. Graham R. C., Jr, Karnovsky M. J., Shafer A. W., Glass E. A., Karnovsky M. L. Metabolic and morphological observations on the effect of surface-active agents of leukocytes. J Cell Biol. 1967 Mar;32(3):629–647. doi: 10.1083/jcb.32.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hellum K. B., Solberg C. O. Influence of anticoagulants on the nitroblue tetrazolium test. Scand J Infect Dis. 1973;5(1):67–70. doi: 10.3109/inf.1973.5.issue-1.12. [DOI] [PubMed] [Google Scholar]
  6. Holmes B., Page A. R., Good R. A. Studies of the metabolic activity of leukocytes from patients with a genetic abnormality of phagocytic function. J Clin Invest. 1967 Sep;46(9):1422–1432. doi: 10.1172/JCI105634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. LAURELL C. B. ANTIGEN-ANTIBODY CROSSED ELECTROPHORESIS. Anal Biochem. 1965 Feb;10:358–361. doi: 10.1016/0003-2697(65)90278-2. [DOI] [PubMed] [Google Scholar]
  8. Matula G., Paterson P. Y. Spontaneous in vitro reduction of nitroblue tetrazolium by neutrophils of adult patients with bacterial infection. N Engl J Med. 1971 Aug 5;285(6):311–317. doi: 10.1056/NEJM197108052850603. [DOI] [PubMed] [Google Scholar]
  9. McCall C. E., DeChatelet L. R., Cooper M. R., Shannon C. Human toxic neutrophils. 3. Metabolic characteristics. J Infect Dis. 1973 Jan;127(1):26–33. doi: 10.1093/infdis/127.1.26. [DOI] [PubMed] [Google Scholar]
  10. McCall C. E., Katayama I., Cotran R. S., Finland M. Lysosomal and ultrastructural changes in human "toxic" neutrophils during bacterial infection. J Exp Med. 1969 Feb 1;129(2):267–293. doi: 10.1084/jem.129.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nathan D. G., Baehner R. L., Weaver D. K. Failure of nitro blue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease. J Clin Invest. 1969 Oct;48(10):1895–1904. doi: 10.1172/JCI106156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Park B. H., Fikrig S. M., Smithwick E. M. Infection and nitroblue-tetrazolium reduction by neutrophils. A diagnostic acid. Lancet. 1968 Sep 7;2(7567):532–534. doi: 10.1016/s0140-6736(68)92406-9. [DOI] [PubMed] [Google Scholar]
  13. Segal A. W., Levi A. J. The mechanism of the entry of dye into neutrophils in the nitroblue tetrazolium (NBT) test. Clin Sci Mol Med. 1973 Dec;45(6):817–826. doi: 10.1042/cs0450817. [DOI] [PubMed] [Google Scholar]
  14. Stossel T. P. Evaluation of opsonic and leukocyte function with a spectrophotometric test in patients with infection and with phagocytic disorders. Blood. 1973 Jul;42(1):121–130. [PubMed] [Google Scholar]
  15. Ward P. A., Zvaifler N. J. Quantitative phagocytosis by neutrophils. I. A new method with immune complexes. J Immunol. 1973 Dec;111(6):1771–1776. [PubMed] [Google Scholar]

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