Witko-Sarsat V, Nguyen A T, Knight J, Descamps-Latscha B
INSERM U25, Hôpital Necker, Paris, France.
Free Radic Biol Med. 1992;13(1):83-8. doi: 10.1016/0891-5849(92)90169-h.
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).
我们之前报道过，正常人多形核中性粒细胞（PMN）在存在外源性牛磺酸的情况下，受到佛波酯（PMA）或调理酵母聚糖刺激时会释放牛磺酸氯胺，一种寿命较长的氧化剂。我们现在描述一种新的、简单且高度灵敏的检测氯胺（包括牛磺酸氯胺）的方法，该方法使用化学发光探针Pholasin，即软体动物大西洋船蛆（Pholas dactylus）的荧光素。采用比色法（基于碘化钾的氧化）和依赖Pholasin的化学发光（CL）法评估牛磺酸氯胺（N - 氯代牛磺酸）的检测情况。使用碘化钾（KI）法测定的PMN上清液中牛磺酸氯胺浓度与依赖Pholasin的CL密切相关。还在非氧化条件下评估了这种CL。使用碘化钾（KI）法在患有氧化爆发缺陷（慢性肉芽肿病，CGD）患者的淋巴细胞和PMN上清液中未检测到牛磺酸氯胺，但持续观察到依赖Pholasin的CL。在我们的孵育条件下，使用甲硫氨酸（一种特定的氯胺清除剂）使我们能够定义依赖Pholasin的CL中可被甲硫氨酸抑制的部分（即氯胺诱导的CL）。
