Johnston R B, Godzik C A, Cohn Z A
J Exp Med. 1978 Jul 1;148(1):115-27. doi: 10.1084/jem.148.1.115.
We studied the capacity of cultured mouse peritoneal macrophages to generate superoxide anion (O(2-)), the initial product of conversion of oxygen to microbicidal species, during phagocytosis of opsonized zymosan or upon contact with the membrane-active agent phorbel myristate acetate (PMA). Macrophages from mice infected with Bacille Calmette-Guerin (BCG) or injected intraperitoneally with thioglycollate broth or endotoxin, released up to 12 times more O(2-) than did resident peritoneal macrophages, depending upon the cell type and whether the stimulus was zymosan or PMA. There was little if any O(2-) release from resting (unstimulated) macrophages. The density of cells on culture dishes was an important variable since crowding of the dish markedly reduced the efficiency of O(2-) production. The enhanced O(2-) release of chemically elicited and infection-activated macrophages was noted after stimulation with a wide range of concentrations of PMA and zymosan, at all time points studied (up to 120 min), and with cells maintained for 140 rain to 16 days in culture. The O(2-) response of resident cells improved twofold to zymosan and ninefold to PMA during the first 3 days in culture. The capacity to release O~ appears to be limited to actively phagocytic cell types: murine macrophage-like tumor lines and cultured human monocytes released O(2-) when stimulated by PMA or zymosan, fibroblast and endothelial lines and embryo-derived cells did not. Activity of superoxide dismutase, which removes O(2-), was not detectable in culture supernates of any cell type, and thus, differences in detectable O(2-) could not be attributed to variations in the release of this enzyme. We conclude that the phagocytosis- associated respiratory burst is significantly enhanced in mononuclear phagocytes obtained ai~r chemical inflammation or BCG infection. Increased capacity to generate O(2-) and other oxygen radicals during phagocytosis could contribute to the improved microbicidal and tumoricidal activity of activated macrophages.
我们研究了培养的小鼠腹腔巨噬细胞在吞噬调理酵母聚糖或与膜活性剂佛波酯肉豆蔻酸酯乙酸酯（PMA）接触过程中产生超氧阴离子（O₂⁻）的能力，O₂⁻是氧气转化为杀菌物质的初始产物。感染卡介苗（BCG）或腹腔注射巯基乙酸肉汤或内毒素的小鼠的巨噬细胞，根据细胞类型以及刺激物是酵母聚糖还是PMA，释放的O₂⁻比常驻腹腔巨噬细胞多高达12倍。静息（未刺激）的巨噬细胞几乎不释放O₂⁻。培养皿上细胞的密度是一个重要变量，因为培养皿拥挤会显著降低O₂⁻产生的效率。在研究的所有时间点（长达120分钟），用各种浓度的PMA和酵母聚糖刺激后，以及培养140分钟至16天的细胞中，都观察到化学诱导和感染激活的巨噬细胞O₂⁻释放增强。培养的前3天，常驻细胞对酵母聚糖的O₂⁻反应提高了两倍，对PMA的反应提高了九倍。释放O₂⁻的能力似乎仅限于活跃吞噬的细胞类型：小鼠巨噬细胞样肿瘤细胞系和培养的人单核细胞在受到PMA或酵母聚糖刺激时释放O₂⁻，成纤维细胞、内皮细胞系和胚胎来源的细胞则不释放。在任何细胞类型的培养上清液中均未检测到去除O₂⁻的超氧化物歧化酶的活性，因此，可检测到的O₂⁻差异不能归因于该酶释放的变化。我们得出结论，在化学炎症或BCG感染后获得的单核吞噬细胞中，与吞噬相关的呼吸爆发显著增强。吞噬过程中产生O₂⁻和其他氧自由基的能力增强可能有助于提高活化巨噬细胞的杀菌和杀肿瘤活性。
