Killing of intracellular Leishmania donovani by human mononuclear phagocytes. Evidence for oxygen-dependent and -independent leishmanicidal activity.
Killing of intracellular Leishmania donovani by human mononuclear phagocytes. Evidence for oxygen-dependent and -independent leishmanicidal activity.
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人单核吞噬细胞杀死细胞内杜氏利什曼原虫。
DOI：
10.1172/jci110972
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Cartelli,DM
中科院分区：
文献类型：
--
作者：
Murray,HW;Cartelli,DM
在检测 H2O2 释放或用杜氏利什曼原虫前鞭毛体 (LDP) 或无鞭毛体 (LDA) 攻击之前，培养人外周血单核细胞 1-30 天。 1-d 细胞响应佛波醇肉豆蔻酸酯乙酸酯触发（1,013 +/- 58 nmol/mg 蛋白质。90 分钟）和 LDP 摄入，很容易产生 H2O2，并在 6 小时内杀死 50% 的 LDP，到 24 小时杀死 90%。相比之下，相同的细胞在摄入LDA期间释放很少的H2O2，在6小时时没有杀死LDA，到24小时时释放的H2O不到30%，并且支持细胞内LDA复制。单核细胞衍生的巨噬细胞（首先培养大于或等于 7 天的细胞）产生的 H2O2 低于 125 nmol/mg。佛波醇肉豆蔻酸酯乙酸酯触发后 90 分钟，既不杀死 LDP 也不杀死 LDA，并且允许两种形式复制。然而，添加有丝分裂原或抗原刺激的淋巴因子可以防止单核细胞氧化能力的下降，使巨噬细胞 H2O2 释放增加六倍以上，同时诱导 1-d 单核细胞杀死 LDA，并培养巨噬细胞显示杀鞭毛体和杀鞭毛活性。与正常供体的 1 天单核细胞和淋巴因子激活的巨噬细胞相比，来自慢性肉芽肿病 (CGD) 患者的相同细胞或氧化活性因过氧化氢酶预处理或葡萄糖剥夺而受损的正常细胞在吞噬后早期（6-24 小时）表现出相当低的抗利什曼活性，或没有表现出抗利什曼活性。然而，感染后 48 小时，1-d CGD 单核细胞和氧化受损的正常细胞分别杀死了 40% 和超过 80% 的 LDP。尽管需要较长时间的淋巴因子刺激，并且所产生的抗利什曼效应不如正常细胞那么快，但活化的CGD单核细胞和巨噬细胞最终也实现了杀鞭毛和抑制鞭毛的活性。这些结果表明，人类单核吞噬细胞利用氧依赖性和非依赖性机制来实现针对摄入的利什曼原虫的活性，并且还证明了（a）两种形式的杜氏利什曼原虫对细胞内杀伤的不同敏感性，（b）氧中间体在有效的单核吞噬细胞抗菌活性中的关键作用，（c）淋巴细胞产物增强氧依赖性和非依赖性途径的能力，以及（d）在没有淋巴因子刺激的情况下，单核细胞来源的巨噬细胞对利什曼原虫感染的影响。图像图2
Human peripheral blood monocytes were cultivated for 1-30 d before assay for H2O2 release or challenge with Leishmania donovani promastigotes (LDP) or amastigotes (LDA). 1-d cells readily generated H2O2 in response to both phorbol myristate acetate triggering (1,013 +/- 58 nmol/mg protein . 90 min) and LDP ingestion, and killed 50% of LDP within 6 h, and 90% by 24 h. In contrast, the same cells released little H2O2 during LDA ingestion, killed no LDA at 6 h and less than 30% by 24 h, and supported intracellular LDA replication. Monocyte-derived macrophages (cells first cultivated for greater than or equal to 7 d) generated less than 125 nmol H2O2/mg . 90 min after phorbol myristate acetate triggering, killed neither LDP nor LDA, and permitted both forms to replicate. The addition of mitogen- or antigen-stimulated lymphokines, however, prevented the decline in monocyte oxidative capacity, enhanced macrophage H2O2 release by more than sixfold, and, in parallel, induced 1-d monocytes to kill LDA and cultivated macrophages to display both promastigocidal and amastigocidal activity. In comparison to 1-d monocytes and lymphokine-activated macrophages from normal donors, the same cells from patients with chronic granulomatous disease (CGD) or normal cells whose oxidative activity had been impaired by catalase pretreatment or glucose deprivation exerted considerably less or no antileishmanial activity during the early (6-24 h) postphagocytic period. By 48 h after infection, however, 1-d CGD monocytes and oxidatively impaired normal cells killed 40 and greater than 80% of LDP, respectively. Although a longer period of lymphokine stimulation was required and the resulting antileishmanial effects were not as rapid as with normal cells, activated CGD monocytes and macrophages also eventually achieved promastigocidal and amastigostatic activity. These results indicate that human mononuclear phagocytes utilize both oxygen-dependent and -independent mechanisms to achieve activity against ingested Leishmania, and also demonstrate (a) the differential susceptibilities of the two forms of L. donovani to intracellular killing, (b) the key role of oxygen intermediates in effective mononuclear phagocyte antimicrobial activity, (c) the capacity of lymphocyte products to enhance oxygen-dependent as well as -independent pathways, and (d) the vulnerability of the monocyte-derived macrophage to Leishmania infection in the absence of lymphokine stimulation.ImagesFIGURE 2