Witko V, Nguyen A T, Descamps-Latscha B
INSERM U25, Hôpital Necker, Paris, France.
J Clin Lab Anal. 1992;6(1):47-53. doi: 10.1002/jcla.1860060110.
Despite their potential importance, the role of phagocyte-derived chloramines ("long-lived oxidants") has not yet been investigated in inflammatory or infectious diseases. We have developed a sensitive spectrophotometric microtiter plate assay for chloramines based on their capacity to oxidize potassium iodide (KI). Consistent levels of endogenous chloramines were detected in normal human polymorphonuclear neutrophil (PMN) supernatants after stimulation by phorbol myristate acetate (PMA) or opsonized zymosan. Exogenous taurine strongly enhanced chloramine secretion and was used to quantify the chlorinating potential of PMN. Taurine-chloramines were also detectable in monocyte supernatants, although in smaller amounts. The specificity of the KI assay was assessed both in terms of effect of compounds inhibiting (KBr) or interacting with (sodium azide and catalase) chloramine formation and by showing that PMN from patients with chronic granulomatous disease, due to a hereditary lack of oxidative response capacity, were unable to produce chloramines. Taurine-chloramine levels secreted by PMA (but not zymosan)-stimulated PMN were closely related to the cellular luminol-amplified chemiluminescence (CL) responses although the CL assay failed to detect chloramines in PMN supernatants. We consider that this KI assay should be of use in studying the role of long-lived phagocyte-derived oxidants in clinical medicine.
尽管吞噬细胞衍生的氯胺（“长寿命氧化剂”）具有潜在的重要性，但尚未在炎症或感染性疾病中对其作用进行研究。我们基于氯胺氧化碘化钾（KI）的能力，开发了一种灵敏的分光光度微孔板法来检测氯胺。在用佛波酯肉豆蔻酸酯（PMA）或调理酵母聚糖刺激后，在正常人多形核中性粒细胞（PMN）上清液中检测到了内源性氯胺的稳定水平。外源性牛磺酸强烈增强了氯胺的分泌，并用于量化PMN的氯化潜力。在单核细胞上清液中也可检测到牛磺酸 - 氯胺，不过含量较少。通过抑制氯胺形成的化合物（KBr）或与氯胺形成相互作用的化合物（叠氮化钠和过氧化氢酶）的作用，以及通过显示由于遗传性氧化反应能力缺乏导致的慢性肉芽肿病患者的PMN无法产生氯胺，来评估KI测定法的特异性。PMA（而非酵母聚糖）刺激的PMN分泌的牛磺酸 - 氯胺水平与细胞鲁米诺增强的化学发光（CL）反应密切相关，尽管CL测定法未能检测到PMN上清液中的氯胺。我们认为这种KI测定法在研究长寿命吞噬细胞衍生的氧化剂在临床医学中的作用方面应该是有用的。
