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Measurement of human neutrophil respiratory burst activity during phagocytosis of bacteria
Book chapter   Peer reviewed

Measurement of human neutrophil respiratory burst activity during phagocytosis of bacteria

Richard F. Rest
Methods in Enzymology
1994
PMID: 7968605

Abstract

This chapter describes the methods to measure the level of human neutrophil respiratory burst activity induced in response to bacterial challenge. Phagocyte activation can occur in the presence or absence of phagocytosis, if there is contact between bacteria and phagocytes. Although similar results may be obtained using granulocytes from animals, they can differ dramatically, both structurally and functionally, from human neutrophils. The methods are presented as a set of assays to begin an organized approach to measure the oxidative burst induced by the bacterial strain(s) under investigation in the laboratory. Each bacterial strain yields slightly different results, and major differences might be observed using unopsonized versus opsonized bacteria. The methods presented include assays to measure two afferent and several efferent activities of the oxidative burst, including oxygen uptake, hexose monophosphate shunt activity, intracellular and extracellular production of superoxide, intracellular and extracellular production of hydrogen peroxide, luminol- and lucigenin-dependent chemiluminescence, and nitroblue tetrazolium (NBT) reduction.

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Web of Science research areas
Biochemical Research Methods
Biochemistry & Molecular Biology
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