Publication: Nitric oxide production by human peripheral blood mononuclear cells
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Issued Date
2001-05-01
Resource Type
ISSN
03044602
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2-s2.0-0035346046
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Mahidol University
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SCOPUS
Bibliographic Citation
Annals of the Academy of Medicine Singapore. Vol.30, No.3 (2001), 270-273
Suggested Citation
P. Toomtong, J. D. Young Nitric oxide production by human peripheral blood mononuclear cells. Annals of the Academy of Medicine Singapore. Vol.30, No.3 (2001), 270-273. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/26798
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Title
Nitric oxide production by human peripheral blood mononuclear cells
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Abstract
Introduction: There are conflicting data on the ability of human mononuclear cells to produce nitric oxide (NO). We investigated nitric oxide production from peripheral blood mononuclear cells (PBMs) by using a new sensitive fluorescent indicator. Materials and Methods: PBMs from healthy volunteers were collected, plated in 96-well microplates, and loaded with the fluorescent nitric oxide probe, 4,5-diaminofluorescein diacetate (DAF-2DA). Experiments were performed in normal control and endotoxin-stimulated PBMs, with and without exogenous L-arginine. The exogenous nitric oxide donor S-nitroso-N-acetyl-penicillamine (SNAP) was used as a positive control. Fluorescence intensity was measured with a fluorescence microplate reader. Results: Nitric oxide production by human PBMs can be demonstrated by the use of the fluorescent indicator, DAF-2DA, in both control and endotoxin-stimulated conditions. Nitric oxide production was independent of the concentration of exogenous L-arginine. The addition of endotoxin did not change nitric oxide production. PBMs treated with SNAP showed a concentration dependent increase in fluorescence. Nitric oxide production over 5 hours was constant and identical in both control and stimulated groups. Conclusion: This fluorescent indicator technique is useful for the study of NO production by human PBMs. Nitric oxide production by PBMs was independent of exogenous L-arginine concentration and was not affected by endotoxin.