Schmitt M, Cochrane C G
Scripps Clinic and Research Foundation, La Jolla, CA 92037.
Free Radic Res Commun. 1987;2(4-6):359-68. doi: 10.3109/10715768709065303.
Binding of purified monoclonal antibody (moAB) IgM NMS-1 to suspended initially spherical living human PMNLs is not associated with the generation of chemiluminescence but was found to enhance the chemiluminescence response to the N-formyl chemotactic peptide FNLPNTL. We investigated quantitatively the kinetics of oxygen metabolite generation by PMNLs stimulated with FNLPNTL +/- moAB NMS-1 using luminol-dependent chemiluminescence as a very sensitive detection system. Chemiluminescence detection allowed the analysis of the time sequence of onset and development of reactive oxygen metabolites following stimulation of PMNLs by FNLPNTL in the presence of moAB NMS-1. The increase of response of PMNLs stimulated with FNLPNTL in the presence of moAB NMS-1 depended on the concentration of the antibody and the sequence of stimulus addition. Stimulation of human PMNLs by 10 nM FNLPNTL induced a rapid burst of chemiluminescence which peaked approximately 5 min after stimulus addition. The subsequent addition of moAB NMS-1 (greater than or equal to 2 micrograms/ml DPBS(+) - 0.1% HSA, 37 degrees C) to FNLPNTL-stimulated PMNLs - after the FNLPNTL-mediated response had already decayed (16-18 min) - without delay induced a second burst of oxygen metabolite generation. The magnitude of this second peak of activation was dose-dependent. Treatment of PMNLs with moAB NMS-1 (greater than or equal to 1 microgram/ml DPBS(+) - 0.1% HSA, 3 min, 37 degrees C) - prior to FNLPNTL (10 nM) stimulation - increased rate and magnitude of the FNLPNTL-mediated response. This response is biphasic with the first peak at the FNLPNTL position and a second, higher peak approximately 16 min after FNLPNTL addition. The magnitude of response was dose-dependent. The latency (lag time) of the response was not changed compared to controls which received no moAB NMS-1 treatment. The observed moAB NMS-1 dependent increase in FNLPNTL-mediated chemiluminescence is transient (50-60 min), persistent activation was not detected.
纯化的单克隆抗体（moAB）IgM NMS-1与最初呈球形悬浮的活人类中性粒细胞的结合并不伴随着化学发光的产生，但发现它能增强对N-甲酰基趋化肽FNLPNTL的化学发光反应。我们使用鲁米诺依赖性化学发光作为一种非常灵敏的检测系统，定量研究了在有或无moAB NMS-1的情况下，FNLPNTL刺激中性粒细胞产生氧代谢产物的动力学。化学发光检测能够分析在moAB NMS-1存在的情况下，FNLPNTL刺激中性粒细胞后活性氧代谢产物产生的起始和发展的时间顺序。在moAB NMS-1存在的情况下，FNLPNTL刺激的中性粒细胞反应的增强取决于抗体的浓度和刺激添加的顺序。10 nM FNLPNTL刺激人类中性粒细胞会诱导化学发光迅速爆发，在刺激添加后约5分钟达到峰值。在FNLPNTL介导的反应已经衰减（16 - 18分钟）后，随后向FNLPNTL刺激的中性粒细胞中添加moAB NMS-1（≥2微克/毫升DPBS(+) - 0.1% HSA，37℃），会立即诱导氧代谢产物产生的第二次爆发。这次激活的第二个峰值的大小是剂量依赖性的。在FNLPNTL（10 nM）刺激之前，用moAB NMS-1（≥1微克/毫升DPBS(+) - 0.1% HSA，3分钟，37℃）处理中性粒细胞，会增加FNLPNTL介导的反应的速率和幅度。这种反应是双相的，第一个峰值在FNLPNTL作用时出现，第二个更高的峰值在添加FNLPNTL后约16分钟出现。反应的幅度是剂量依赖性的。与未接受moAB NMS-1处理的对照组相比，反应的潜伏期（滞后时间）没有变化。观察到的moAB NMS-1依赖性增强的FNLPNTL介导的化学发光是短暂的（50 - 60分钟），未检测到持续激活。
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