Ignarro L J, George W J
J Exp Med. 1974 Jul 1;140(1):225-38. doi: 10.1084/jem.140.1.225.
The purpose of this investigation was to elucidate the relationship of cyclic GMP and calcium to the immunologic discharge of lysosomal enzymes from purified human neutrophils. Contact of neutrophils with a variety of immunologic stimuli, including zymosan particles treated with either normal or rheumatoid arthritic (RA) serum, heat-aggregated (agg) IgG, particulate and immobilized agg IgG each treated with RA serum, and zymosan-treated serum, provoked the discharge of beta-glucuronidase, but not cytoplasmic lactate dehydrogenase, and stimulated the accumulation of cyclic GMP. Both enzyme release and elevation of cyclic GMP levels required the presence of extracellular calcium as neither cellular event proceeded in its absence. Cholinergic enhancement of the immunologic secretion of beta-glucuronidase from neutrophils by acetylcholine was associated with a concomitant accumulation of cyclic GMP. These actions of acetylcholine on neutrophils did not proceed in the absence of extracellular calcium. Whereas the concentrations of cyclic GMP in neutrophils were elevated by both immune reactants and a combination of the latter and acetylcholine, cyclic AMP levels remained unaltered. Thus, cyclic GMP, but not cyclic AMP, was associated with the immunologic and pharmacologic discharge of lysosomal enzymes from neutrophils. Contrariwise, cyclic AMP, but not cyclic GMP, was associated with inhibition of lysosomal enzyme release. For example, dibutyryl cyclic AMP and epinephrine inhibited the release of beta-glucuronidase from neutrophils that was elicited by each of the immune reactants tested. Moreover, cyclic AMP levels in the cells were elevated markedly in every instance that enzyme discharge was inhibited by epinephrine. Epinephrine did not alter the neutrophil concentrations of cyclic GMP at times when those of cyclic AMP were elevated. The data in this report constitute partial evidence that the immunologic discharge of lysosomal enzymes from human neutrophils is mediated or signaled by intracellular cyclic GMP and that calcium is linked to this stimulation of enzyme secretion.
本研究的目的是阐明环磷酸鸟苷（cGMP）和钙与从纯化的人中性粒细胞中溶酶体酶的免疫释放之间的关系。中性粒细胞与多种免疫刺激物接触，包括用正常或类风湿性关节炎（RA）血清处理的酵母聚糖颗粒、热聚集（agg）IgG、用RA血清处理的颗粒状和固定化agg IgG以及酵母聚糖处理的血清，可引发β-葡萄糖醛酸酶的释放，但不会引发细胞质乳酸脱氢酶的释放，并刺激cGMP的积累。酶释放和cGMP水平升高均需要细胞外钙的存在，因为在没有细胞外钙的情况下这两种细胞事件都不会发生。乙酰胆碱对中性粒细胞β-葡萄糖醛酸酶免疫分泌的胆碱能增强作用与cGMP的同时积累有关。乙酰胆碱对中性粒细胞的这些作用在没有细胞外钙的情况下不会发生。虽然免疫反应物以及后者与乙酰胆碱的组合均可使中性粒细胞中的cGMP浓度升高，但环磷酸腺苷（cAMP）水平保持不变。因此，与中性粒细胞溶酶体酶的免疫和药理释放相关的是cGMP，而非cAMP。相反，与溶酶体酶释放抑制相关的是cAMP，而非cGMP。例如，二丁酰环磷酸腺苷和肾上腺素可抑制测试的每种免疫反应物引发的中性粒细胞β-葡萄糖醛酸酶的释放。此外，在肾上腺素抑制酶释放的每种情况下，细胞内的cAMP水平均显著升高。在cAMP水平升高时，肾上腺素不会改变中性粒细胞中cGMP的浓度。本报告中的数据构成了部分证据，表明人中性粒细胞溶酶体酶的免疫释放是由细胞内cGMP介导或作为信号的，并且钙与这种酶分泌的刺激有关。
