Pholasin: a new chemiluminescent probe for the detection of chloramines derived from human phagocytes.
We have previously reported that normal human polymorphonuclear neutrophils (PMN) release taurine-chloramine, a long-lived oxidant, in response to stimulation by phorbol myristate acetate (PMA) or opsonized zymosan in the presence of exogenous taurine. We now describe a new, simple, and highly sensitive method for the detection of chloramines, including taurine-chloramine, using the chemiluminescent probe Pholasin, the luciferin of the mollusc Pholas dactylus. Taurine-chloramine (N-chlorotaurine) detection was assessed with both a colorimetric method (based on the oxidation of potassium iodide) and with the Pholasin-dependent chemiluminescence (CL) method. The taurine-chloramine concentration in PMN supernatants determined using the potassium iodide (KI) method correlated closely with Pholasin-dependent CL. This CL was also assessed in nonoxidative conditions. No taurine-chloramine was detected in supernatants of lymphocytes and PMN from patients with an oxidative burst defect (chronic granulomatous disease, CGD) with the KI method, but Pholasin-dependent CL was consistently observed. The use of methionine, a specific chloramine scavenger in our incubation conditions, allowed us to define a methionine-inhibitable fraction of Pholasin-dependent CL (i.e., chloramine-induced CL).
Witko-Sarsat
V
V
INSERM U25, Hôpital Necker, Paris, France.
Nguyen
A T
AT
Knight
J
J
Descamps-Latscha
B
B
D002648:Child; D002700:Chloramines / Q000378:metabolism*; D049454:Firefly Luciferin; D005609:Free Radicals; D006105:Granulomatous Disease, Chronic / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D009504:Neutrophils / Q000378:metabolism; D010586:Phagocytes / Q000378:metabolism*; D013654:Taurine / Q000031:analogs & derivatives / Q000378:metabolism
D002700:Chloramines; D005609:Free Radicals; D007202:Indicators and Reagents; D013654:Taurine; C043410:N-chlorotaurine; D049454:Firefly Luciferin; C052157:pholasin