| Summary: | Immune information in the form of inflammatory mediators directs phagocyte locomotion and increases expression of opsonin receptors such that contact with an opsonized microbe results in receptor ligation and activation of microbicidal metabolism. Carbohydrate dehydrogenation and O2 consumption feed reactions that effectively lower the spin quantum number (S) of O2 from 1 to 1/2 and finally to 0. Oxidase-catalyzed univalent reduction of O2 (S = 1; triplet multiplicity) yields hydrodioxylic acid (HO2) and its conjugate base superoxide, O2- (S = 1/2; doublet multiplicity). Acid or enzymatic disproportionation of superoxide yields H2O2 (S = 0; singlet multiplicity). Haloperoxidase catalyzes H2O2-dependent oxidation of Cl- yielding HOCl (S = 0), and reaction of HOCl with H2O2 yields singlet molecular oxygen, 1O2 (S = 0; singlet multiplicity). The Wigner spin conservation rule restricts direct reaction of S = 1 O2 with S = 0 organic molecules. Lowering the S of O2 overcomes this spin restriction and allows microbicidal combustion. High exergonicity dioxygenation reactions yield electronically excited carbonyl products that relax by photon emission, i.e., phagocyte luminescence. Addition of high quantum yield substrates susceptible to spin allowed dioxygenation, i.e., chemiluminigenic substrates, greatly increases detection sensitivity and defines the nature of the oxygenating agent. Measurement of luminescence allows high sensitivity, real-time, and substrate-specific differential analysis of phagocyte dioxygenating activities. Under assay conditions where immune mediator and opsonin exposure are controlled, luminescence analysis of the initial phase of opsonin-stimulated oxygenation activity allows functional assessment of the opsonin receptor expression per circulating phagocyte and can be used to gauge the in vivo state of immune activation.
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