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Optimization of peroxide production by resident macrophages
Author(s) -
CookMills Joan M.,
Fraker Pamela J.
Publication year - 1993
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.53.2.205
Subject(s) - biology , production (economics) , peroxide , microbiology and biotechnology , macrophage , immunology , biochemistry , in vitro , chemistry , organic chemistry , macroeconomics , economics
Previous reports indicated that resident macrophages produced low or nondetectable quantities of H2O2 (1—10 nmol H2O2/mg resident macrophage protein) when measured by the phenol red assay described by Pick and Mizel [5]. However, modifications of the assay that included addition of calcium to assay solutions, alterations in cell‐harvesting methods, increasing the concentration of cells analyzed, incubating the cells in ambient air, and enhancing the interactions between macrophages and particulate stimulants by centrifugation resulted in increased production of peroxide. Each variable was instrumental in improving the assay conditions and the combination of changes dramatically increased the amount of peroxide (30‐60 nmol H2O2/mg resident macrophage protein) obtained from murine resident macrophages, making it possible to better assess the role of these cells in microbicidal killing.