Polymorphonuclear neutrophils (PMN) have a remarkable capacity for generation of large amounts of
reactive oxygen species in response to a variety of infectious or inflammatory stimuli, a process known as the respiratory burst that involves activation of a multicomponent
NADPH oxidase. Given their short life span, PMN are not amenable to most molecular biology methods for studying activation of this
oxidant-generating system. We have explored a variety of methods for introduction of components of the phagocytic
oxidase (
phox system) into the promyelocytic
erythroleukemia cell line, K-562. Here, we describe a series of cloned K-562 cell lines that were retrovirally transduced for stable production of one or more essential components of the phagocytic
oxidase (
phox) complex. We outline methods for the use of these transfectable cells for investigating structure, function, and signaling requirements for assembly and activation of the
phox system. These versatile lines can be used to examine effects of genetic polymorphisms or mutations in
phox components associated with
chronic granulomatous disease, to serve as a system for testing gene therapy vectors designed to correct the defective
oxidase, to study cross-functioning with recently described
phox component homologs, or to explore signaling components involved in regulation of the respiratory burst.