Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs: A mechanism of toxicity

R. A. Cassidy, D. G. Burleson, A. V. Delgado, D. J. Kohler, M. L. Salin, Basil A Pruitt

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Isolated human PMNs served as a model to determine oxyhemoglobin (oxyHb) binding and the effects of oxymyoglobin (oxyMb) or oxyHb on production of both nitric oxide (NO·) and superoxide (O2·-) and the resulting cytotoxicity. Physiologically relevant concentrations of NO· and H2O2 oxidized, to a similar extent, 2,7-dichlorodihydro-fluorescein (DCFH) loaded into polymorphonuclear neutrophils (PMNs). Activation of PMNs with phorbol 12-myristate 13-acetate (PMA) markedly increased the internalization of extracellular oxyHb (10-250 μg/mL). OxyMb (10-300 μg/mL) or oxyHb (30-300 μg/mL) enhanced DCFH oxidation by a concentration-dependent mechanism in unstimulated, lipopolysaccharide (LPS) and tumor necrosis factor α (TNF-α)-, and PMA-stimulated PMNs. This increased DCFH oxidation was eliminated by NO· synthase inhibitors, glutathione and ascorbate, and was reduced by albumin. Nitrite accumulation in PMN filtrates mirrored NO·-induced DCF fluorescence. OxyMb-induced increases in NO· levels paralleled alterations in DNA and cell membrane damage and ATP levels in PMNs and co-cultured lymphocytes, and were attenuated by NO· synthase inhibitors. OxyMb eliminated extracellular O2·- at protein concentrations 100- to 1000-fold above those of superoxide dismutase. These results suggest that heme proteins bind and internalize into PMNs and increase NO·-induced damage in neighboring cells by inhibiting O2·--scavenging of NO·. We propose a mechanism whereby heme protein-induced NO· levels may contribute to immunosuppression and increased infection rates associated with transfusions and cellular damage during inflammation.

Original languageEnglish (US)
Pages (from-to)357-368
Number of pages12
JournalJournal of Leukocyte Biology
Volume67
Issue number3
StatePublished - 2000
Externally publishedYes

Fingerprint

Hemeproteins
Cell Survival
Nitric Oxide
Neutrophils
Oxyhemoglobins
Nitric Oxide Synthase
Acetates
Neutrophil Activation
Nitrites
Fluorescein
Superoxides
Immunosuppression
Superoxide Dismutase
Glutathione
Lipopolysaccharides
Albumins
Tumor Necrosis Factor-alpha
Adenosine Triphosphate
Fluorescence
Cell Membrane

Keywords

  • Apoptosis
  • Cytotoxicity
  • DNA strand breaks
  • Hemoglobin
  • Myoglobin
  • Superoxide

ASJC Scopus subject areas

  • Cell Biology

Cite this

Cassidy, R. A., Burleson, D. G., Delgado, A. V., Kohler, D. J., Salin, M. L., & Pruitt, B. A. (2000). Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs: A mechanism of toxicity. Journal of Leukocyte Biology, 67(3), 357-368.

Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs : A mechanism of toxicity. / Cassidy, R. A.; Burleson, D. G.; Delgado, A. V.; Kohler, D. J.; Salin, M. L.; Pruitt, Basil A.

In: Journal of Leukocyte Biology, Vol. 67, No. 3, 2000, p. 357-368.

Research output: Contribution to journalArticle

Cassidy, RA, Burleson, DG, Delgado, AV, Kohler, DJ, Salin, ML & Pruitt, BA 2000, 'Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs: A mechanism of toxicity', Journal of Leukocyte Biology, vol. 67, no. 3, pp. 357-368.
Cassidy RA, Burleson DG, Delgado AV, Kohler DJ, Salin ML, Pruitt BA. Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs: A mechanism of toxicity. Journal of Leukocyte Biology. 2000;67(3):357-368.
Cassidy, R. A. ; Burleson, D. G. ; Delgado, A. V. ; Kohler, D. J. ; Salin, M. L. ; Pruitt, Basil A. / Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs : A mechanism of toxicity. In: Journal of Leukocyte Biology. 2000 ; Vol. 67, No. 3. pp. 357-368.
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