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Related Experiment Videos

N-nitrosamine formation by macrophages.

M Miwa1, D J Stuehr, M A Marletta

  • 1Massachusetts Institute of Technology, Department of Applied Biological Sciences, Cambridge 02139.

IARC Scientific Publications
|January 1, 1987
PubMed
Summary

Stimulated macrophages can form N-nitrosamines, such as N-nitrosomorpholine (NMOR), when exposed to lipopolysaccharide (LPS) and morpholine. This finding suggests macrophages may produce nitrosamines under certain physiological conditions.

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Area of Science:

  • Immunology
  • Biochemistry
  • Toxicology

Background:

  • Nitrate biosynthesis is a recognized mammalian process.
  • Macrophages stimulated with Escherichia coli lipopolysaccharide (LPS) synthesize nitrate.
  • Macrophages produce both nitrite and nitrate in cell cultures.

Purpose of the Study:

  • To investigate N-nitrosamine formation by stimulated macrophages.
  • To determine if macrophages can produce N-nitrosomorpholine (NMOR).
  • To explore the role of LPS and gamma-interferon (IFN) in this process.

Main Methods:

  • Macrophage cell lines (J774.1, WEHI-3, RAW 264) were cultured.
  • Nitrate concentrations were measured in the supernatant.
  • N-nitrosamines were extracted and analyzed using gas chromatography-thermal energy analysis.

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  • Cells were incubated with LPS, IFN, and morpholine under various conditions.

    • Main Results:

    • LPS-stimulated macrophages produced N-nitrosomorpholine (NMOR) in the presence of morpholine.
    • Both RAW 264 and WEHI-3 cell lines produced NMOR, requiring LPS.
    • gamma-Interferon (IFN) enhanced both NMOR and nitrite formation.
    • Nitrite alone did not form NMOR with morpholine.
    • NMOR formation peaked earlier than nitrite accumulation.

    Conclusions:

    • Macrophages are capable of forming N-nitrosamines under specific stimulated conditions.
    • LPS is crucial for both nitrite and NMOR production in macrophages.
    • The kinetics suggest a complex pathway for nitrosamine formation distinct from simple nitrite conversion.