Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker

PP Bradley, DA Priebat, RD Christensen… - Journal of investigative …, 1982 - Elsevier
PP Bradley, DA Priebat, RD Christensen, G Rothstein
Journal of investigative dermatology, 1982Elsevier
We examined the hypothesis that myeloperoxidase (MPO), a plentiful constituent of
neutrophils, might seve as a marker for tissue neutrophil content. To completely extract MPO
from either neutrophils or skin, hexadecyl-trimethylammonium bromide (HTAB) was used to
solubilize the enzyme. With this detergent treatment, 97.8±0.2% of total recoverable MPO
was extracted from neutrophils with a single HTAB treatment; 93.1±1.0% was solubilized
with a single treatment of skin. Neutrophil MPO was directly related to neutrophil number; …
We examined the hypothesis that myeloperoxidase (MPO), a plentiful constituent of neutrophils, might seve as a marker for tissue neutrophil content. To completely extract MPO from either neutrophils or skin, hexadecyl-trimethylammonium bromide (HTAB) was used to solubilize the enzyme. With this detergent treatment, 97.8 ± 0.2% of total recoverable MPO was extracted from neutrophils with a single HTAB treatment; 93.1 ± 1.0% was solubilized with a single treatment of skin. Neutrophil MPO was directly related to neutrophil number; with the dianisidine-H2O2 assay as few as 104 neutrophils could be detected. The background level of MPO within uninflamed tissue was 0.385 ± 0.018 units per gram of tissue, equivalent to only 7.64 ± 0.36 × 105 neutrophils. In experimental staphylococcal infection, skin specimens contained 34.8 ± 3.8 units MPO per gram, equivalent to 8.55 ± 0.93 × 107 neutrophils. These studies demonstrate that MPO can be used as a marker for skin neutrophil content: it is recoverable from skin in soluble form, and is directly related to neutrophil number. Further, normal skin possesses a low background of MPO compared to that of inflamed skin.
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