Skip to main content

The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes.

Publication ,  Journal Article
Johnston, RB; Keele, BB; Misra, HP; Lehmeyer, JE; Webb, LS; Baehner, RL; RaJagopalan, KV
Published in: J Clin Invest
June 1975

The capacity of human phagocytes to generate superoxide anion (O2-), a free radical of oxygen, and a possible role for this radical or its derivatives in the killing of phagocytized bacteria were explored using leukocytes from normal individuals and patients with chronic granulomatous disease (CGD). Superoxide dismutase, which removes O2-, consistently inhibited phagocytosis-associated nitroblue tetrazolium (NBT) reduction indicating the involvement of O2- in this process. Similarly, superoxide dismutase inhibited the luminescence that occurs with phagocytosis, implicating O2- in this phenomenon, perhaps through its spontaneous dismutation into singlet oxygen. Subcellular fractions from homogenates of both normal and CGD leukocytes generated O2- effectively in the presence of NADH as substrate. However, O2- generation by intact cells during phagocytosis was markedly diminished in nine patients with CGD. Leukocytes from mothers determined to be carriers of X-linked recessive CGD by intermediate phagocytic reduction of NBT elaborated O2- to an intermediate extent, further demonstrating the interrelationship between NBT reduction and O2- generation in phagocytizing cells. Activity of superoxide dismutase, the enzyme responsible for protecting the cell from the damaging effects of O2-, was approximately equal in homogenates of normal and CGD granulocytes. Polyacrylamide electrophoresis separated this activity into a minor band that appeared to be the manganese-containing superoxide dismutase associated with mitochondria and a more concentrated, cyanide-sensitive, cytosol form of the enzyme with electrophoretic mobility that corresponded to that of erythrocyte cuprozinc superoxide dismutase. Superoxide dismutase inhibited the phagocytic killing of Escherichia coli, Staphylococcus aureus, and Streptococcus viridans. A similar inhibitory effect was noted with catalase which removes hydrogen peroxide. Neither enzyme inhibited the ingestion of bacteria. Peroxide and O2- are believed to interact to generate the potent oxidant, hydroxyl radical (.OH). A requirement for .OH in the phagocytic bactericidal event might explain the apparent requirement for both O2- and H2O2 for such activity. In agreement with this possibility, benzoate and mannitol, scavengers of .OH, inhibited phagocytic bactericidal activity. Generation of singlet oxygen from O2- and .OH also might explain these findings. It would seem clear from these and other studies that the granulo cyte elaborates O2- as a concomitant of the respiratory burst that occurs with phagocytosis. To what extent the energy inherent in O2- is translated into microbialdeath through O2- itself, hydrogen peroxide, .OH, singlet oxygen, or some other agent remains to be clearly defined.

Duke Scholars

Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

June 1975

Volume

55

Issue

6

Start / End Page

1357 / 1372

Location

United States

Related Subject Headings

  • Superoxides
  • Superoxide Dismutase
  • Streptococcus
  • Staphylococcus
  • Phagocytosis
  • Phagocytes
  • Phagocyte Bactericidal Dysfunction
  • Oxygen
  • Nitroblue Tetrazolium
  • Microscopy, Electron
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Johnston, R. B., Keele, B. B., Misra, H. P., Lehmeyer, J. E., Webb, L. S., Baehner, R. L., & RaJagopalan, K. V. (1975). The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes. J Clin Invest, 55(6), 1357–1372. https://doi.org/10.1172/JCI108055
Johnston, R. B., B. B. Keele, H. P. Misra, J. E. Lehmeyer, L. S. Webb, R. L. Baehner, and K. V. RaJagopalan. “The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes.J Clin Invest 55, no. 6 (June 1975): 1357–72. https://doi.org/10.1172/JCI108055.
Johnston RB, Keele BB, Misra HP, Lehmeyer JE, Webb LS, Baehner RL, et al. The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes. J Clin Invest. 1975 Jun;55(6):1357–72.
Johnston, R. B., et al. “The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes.J Clin Invest, vol. 55, no. 6, June 1975, pp. 1357–72. Pubmed, doi:10.1172/JCI108055.
Johnston RB, Keele BB, Misra HP, Lehmeyer JE, Webb LS, Baehner RL, RaJagopalan KV. The role of superoxide anion generation in phagocytic bactericidal activity. Studies with normal and chronic granulomatous disease leukocytes. J Clin Invest. 1975 Jun;55(6):1357–1372.

Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

June 1975

Volume

55

Issue

6

Start / End Page

1357 / 1372

Location

United States

Related Subject Headings

  • Superoxides
  • Superoxide Dismutase
  • Streptococcus
  • Staphylococcus
  • Phagocytosis
  • Phagocytes
  • Phagocyte Bactericidal Dysfunction
  • Oxygen
  • Nitroblue Tetrazolium
  • Microscopy, Electron