Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity.
Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity.
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DOI：
10.1084/jem.158.3.670
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发表时间：
1983-09-01
期刊：
影响因子：
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通讯作者：
Rubin BY
中科院分区：
文献类型：
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作者：
Nathan CF;Murray HW;Wiebe ME;Rubin BY
经弓形虫抗原、刀豆蛋白A、甲素加小扁豆凝集素或葡萄球菌肠毒素A刺激的人血单核白细胞分泌一种因子(巨噬细胞激活因子或MAF)，增强人巨噬细胞释放过氧化氢和杀灭弓形虫的能力。同样的淋巴上清液中含有干扰素-γ，但不含干扰素-α或干扰素-β。7个未分离上清液中有6个的MAF活性可被中和干扰素-γ的单抗完全消除，其余上清液中的MAF几乎完全中和。经两种相互独立的方案部分纯化的天然干扰素，比活性分别为1×10(6)和10(7)U/mg蛋白，其MAF活性至少与抗病毒活性相同。巨噬细胞在部分纯化的天然干扰素中孵育后分泌过氧化氢的能力(平均峰值刺激，8.8倍)大于未纯化的淋巴因子(3.8倍)，有时相当于或超过新鲜收获的单核细胞的能力。部分纯化的天然干扰素-γ制剂的MAF活性可被单抗-干扰素-γ消除。最后，分离出了纯度大于99%的干扰素-γ(在基因技术公司)。用克隆的人类淋巴因子基因转化的细菌。重组干扰素γ具有较强的MAF活性，在峰值反应时刺激巨噬细胞释放过氧化氢的能力平均提高19.8倍，并将其杀灭弓形虫的能力从未处理细胞的2.6+/-1.3%提高到处理细胞的54+/-0.4%。要达到最大过氧化氢释放能力的50%，需要0.1抗病毒U/ml的重组干扰素的几何平均浓度，估计该制剂的抗病毒浓度约为6微微克分子。巨噬细胞的过氧化分泌量和杀弓形体活性在染毒后2~4d达到高峰。在连续暴露的至少6天内，过氧化物酶的分泌能力保持较高水平，但干扰素-γ的作用在其去除后的大约3天内被逆转。激活通常但不总是伴随着细胞形态的特征性变化。因此，干扰素能激活人巨噬细胞的氧化代谢和抗菌活性，似乎是在所测试的各种LK制剂中唯一能够持续做到这一点的因素。
Human blood mononuclear leukocytes stimulated with toxoplasma antigen, concanavalin A, mezerein plus lentil lectin, or staphylococcal enterotoxin A secreted a factor (macrophage-activating factor, or MAF) that enhanced the capacity of human macrophages to release H2O2 and to kill toxoplasmas. The same lymphoid supernatants contained IFN gamma but not IFN alpha or IFN beta. The MAF activity of six of seven unfractionated supernatants was completely eliminated by a monoclonal antibody that neutralizes IFN gamma, and MAF in the remaining supernatant was almost completely neutralized. Native IFN gamma partially purified by two independent protocols to specific activities of 1 X 10(6) and 10(7) U/mg protein was enriched in MAF activity at least as much as in antiviral activity. The capacity of macrophages to secrete H2O2 after incubation in partially purified native IFN gamma (mean peak stimulation, 8.8-fold) was greater than with unpurified lymphokines (3.8-fold) and sometimes equaled or exceeded the capacity of freshly harvested monocytes. The MAF activity of the partially purified native IFN gamma preparations was abolished by monoclonal anti- IFN gamma. Finally, IFN gamma of greater than 99% estimated purity was isolated (at Genentech, Inc.) from bacteria transformed with the cloned human gene for this lymphokine. Recombinant IFN gamma had potent MAF activity, stimulating the peroxide-releasing capacity of macrophages an average of 19.8-fold at peak response and enhancing their ability to kill toxoplasmas from 2.6 +/- 1.3% for untreated cells to 54 +/- 0.4% for treated cells. Attainment of 50% of the maximal elevation in peroxide-releasing capacity required a geometric mean concentration of 0.1 antiviral U/ml of recombinant IFN gamma, which is estimated to be approximately 6 picomolar for this preparation. Peroxide secretory capacity and toxoplasmacidal activity of macrophages peaked 2-4 d after exposure to IFN gamma. Peroxide-secretory capacity remained elevated during at least 6 d of continuous exposure, but the effect of IFN gamma was reversed within about 3 d of its removal. Activation was usually but not invariably accompanied by characteristic changes in cell morphology. Thus, IFN gamma activates human macrophage oxidative metabolism and antimicrobial activity, and appeared to be the only factor consistently capable of doing so in the diverse LK preparations tested.