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Microtiter plate assay for phagocyte-derived taurine-chloramines
V Witko1, A T Nguyen, B Descamps-Latscha
1INSERM U25, Hôpital Necker, Paris, France.
Journal of Clinical Laboratory Analysis
|January 1, 1992
Summary
Researchers developed a new assay to measure chloramines, which are oxidants produced by immune cells. This method helps study their role in diseases and shows their presence in human neutrophils and monocytes.
Area of Science:
- Immunology
- Biochemistry
Background:
- Phagocyte-derived chloramines, or long-lived oxidants, are crucial in inflammatory and infectious diseases.
- Their role has not been extensively studied due to a lack of appropriate detection methods.
Purpose of the Study:
- To develop and validate a novel assay for detecting and quantifying chloramines.
- To investigate the presence and production of chloramines by human phagocytes.
Main Methods:
- A sensitive spectrophotometric microtiter plate assay was developed using potassium iodide (KI) to detect chloramines.
- The assay's specificity was confirmed using inhibitors and by testing neutrophils from patients with chronic granulomatous disease.
- Chloramine production was stimulated using phorbol myristate acetate (PMA) or opsonized zymosan, with and without exogenous taurine.
Main Results:
- Endogenous chloramines were detected in human polymorphonuclear neutrophil (PMN) and monocyte supernatants.
- Taurine significantly enhanced chloramine secretion, allowing quantification of PMN chlorinating potential.
- The assay demonstrated specificity, and PMN from patients lacking oxidative capacity did not produce chloramines.
- Taurine-chloramine levels correlated with chemiluminescence responses in PMA-stimulated PMN.
Conclusions:
- A novel, sensitive KI-based assay for chloramines has been established.
- This assay facilitates the study of phagocyte-derived oxidants in clinical settings.
- Chloramines are produced by human neutrophils and monocytes and their production can be quantified.