Respiratory Burst in Human Neutrophils

During phagocytosis of microbial intruders, professional phagocytes of our innate immune system increase their oxygen consumption through the activity of an NADPH-oxidase that generates superoxide anion (O(2)(-)) and hydrogen peroxide (H(2)O(2)). These oxygen metabolites give rise to yet other reactive oxygen species that are strongly anti-microbial but which may also cause damage by destructing surrounding tissue and inducing apoptosis in other immune reactive cells. The development of methodology to measure the generation/release of phagocyte respiratory burst products is thus of great importance, and a number of different techniques are currently in use for this purpose. Three of the techniques that we have used, (luminol/isoluminol amplified chemiluminescence, cytochrome C reduction, and PHPA oxidation technique) are described in more detail in this review. We hope to convince the readers that these techniques are valuable tools in basic as well as more clinically oriented research dealing with phagocyte function. The basic principles for luminol/isoluminol-amplified chemiluminescence is used as the starting point for discussing methodological problems related to measurements of oxygen metabolites generated by professional phagocytes.