Devalon M L, Elliott G R, Regelmann W E
University of Minnesota, Department of Pediatrics, Minneapolis 55455.
Infect Immun. 1987 Oct;55(10):2398-403. doi: 10.1128/iai.55.10.2398-2403.1987.
In contrast to results with bacterial suspensions, phagocytosis of unopsonized bacteria readily occurs when bacteria are adhered to glass or plastic surfaces. However, in contrast to neutrophils, alveolar macrophages produced much less DNA denaturation as measured by acridine orange metachromasia of phagocytized Staphylococcus aureus. We have studied the phagocytosis of unopsonized surface-adherent S. aureus and the subsequent production of reactive oxygen species by peripheral blood neutrophils, monocytes, and alveolar macrophages. Phagocyte-free systems were then used to show the relationship of the reactive oxygen species produced by neutrophils and alveolar macrophages and the denaturation of unopsonized S. aureus DNA with acridine orange. Peripheral blood neutrophils, monocytes, and alveolar macrophages from normal human volunteers were added to vials with adherent S. aureus without opsonin. Bacterial uptake and luminol- and lucigenin-dependent chemiluminescence were measured. Neutrophils developed much greater luminol-dependent chemiluminescence than monocytes or alveolar macrophages. Compared with neutrophils and monocytes, alveolar macrophages developed significantly greater concentrations of superoxide, as measured by lucigenin-dependent chemiluminescence and ferricytochrome c reduction. These findings suggested that products of the myeloperoxidase-hydrogen peroxide-halide pathway were generated when peripheral blood neutrophils were stimulated and that alveolar macrophages primarily produced superoxide. When these reactive oxygen species were generated in phagocyte-free systems containing S. aureus, products of the myeloperoxidase-hydrogen peroxide-halide pathway produced denaturation of S. aureus DNA, whereas superoxide did not. Thus, differences in reactive oxygen species produced during phagocytosis may be related to the different capacities of neutrophils and alveolar macrophages to denature unopsonized adherent S. aureus DNA.
与细菌悬液的结果相反，当细菌附着在玻璃或塑料表面时，未调理细菌的吞噬作用很容易发生。然而，与中性粒细胞不同，通过吖啶橙对吞噬的金黄色葡萄球菌进行异染性染色测定，肺泡巨噬细胞产生的DNA变性要少得多。我们研究了未调理的表面附着金黄色葡萄球菌的吞噬作用以及外周血中性粒细胞、单核细胞和肺泡巨噬细胞随后产生活性氧的情况。然后使用无吞噬细胞系统来显示中性粒细胞和肺泡巨噬细胞产生活性氧与未调理的金黄色葡萄球菌DNA用吖啶橙变性之间的关系。将来自正常人类志愿者的外周血中性粒细胞、单核细胞和肺泡巨噬细胞加入到含有未加调理素的附着金黄色葡萄球菌的小瓶中。测量细菌摄取以及鲁米诺和光泽精依赖性化学发光。中性粒细胞产生的鲁米诺依赖性化学发光比单核细胞或肺泡巨噬细胞强得多。通过光泽精依赖性化学发光和高铁细胞色素c还原测定，与中性粒细胞和单核细胞相比，肺泡巨噬细胞产生的超氧化物浓度明显更高。这些发现表明，外周血中性粒细胞受到刺激时会产生活髓过氧化物酶-过氧化氢-卤化物途径的产物，而肺泡巨噬细胞主要产生超氧化物。当在含有金黄色葡萄球菌的无吞噬细胞系统中产生活性氧时，髓过氧化物酶-过氧化氢-卤化物途径的产物会导致金黄色葡萄球菌DNA变性，而超氧化物则不会。因此，吞噬过程中产生活性氧的差异可能与中性粒细胞和肺泡巨噬细胞使未调理的附着金黄色葡萄球菌DNA变性的不同能力有关。
