Abstract
This chapter describes various methods that can be used to measure the activity of nitric oxide synthase (NOS) during inflammation in vivo . These methods demonstrate nitrate synthesis in humans and synthesis from L-[ 15 N] arginine in patients receiving interleukin-2 (IL-2) infusions as cancer immunotherapy. Currently, the role of inducible NOS (iNOS) in human inflammatory states is a controversial issue. This stems primarily from the lack of a reproducible assay for inducing NOS in human mononuclear phagocytes in vitro under conditions where high-output nitric oxide synthesis is observed as in murine macrophages activated with proinflammatory cytokines. iNOS protein—demonstrable by western blot and low-level nitric oxide synthesis—does occur in vitro under defined conditions in mononuclear phagocytes from some individuals. The functional representation of this activity is unknown. The methods described in this chapter are useful for clinical studies toward the understanding the role of NOS in inflammatory diseases, including the biochemical mechanisms operating in cell-mediated immunity to intracellular pathogens in humans.
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