Roach T I, Kiderlen A F, Blackwell J M
Department of Medical Parasitology, London School of Hygiene and Tropical Medicine, United Kingdom.
Infect Immun. 1991 Nov;59(11):3935-44. doi: 10.1128/iai.59.11.3935-3944.1991.
The capacity of mature bone-marrow-derived macrophages and resident peritoneal macrophages from Lshr versus Lshs congenic mice to kill intracellular Leishmania donovani amastigotes when activated by recombinant gamma interferon-lipopolysaccharide (rIFN-gamma-LPS) was examined. IFN-gamma alone in doses up to 100 U/ml was unable to activate macrophages to kill L. donovani amastigotes in vitro; LPS was a necessary secondary stimulus. Similarly, LPS alone in doses up to 100 ng/ml produced no leishmanicidal activity. In bone marrow macrophages, a dose-dependent increase in leishmanicidal activity was observed as increasing rIFN-gamma-LPS dose combinations were introduced, with Lshr macrophages maintaining a significant but not dramatic advantage within any particular dose combination. For peritoneal macrophages, the reverse was true, with macrophages from Lshs mice being more efficient at killing for doses of LPS up to 10 ng/ml with doses of rIFN-gamma in the range of 11 to 33 U/ml. The degree of killing in both bone marrow and peritoneal macrophages correlated well with the levels of nitrites measured in the supernatants at 72 h, and a highly significant correlation was observed between 4-, 24-, or 72-h tumor necrosis factor alpha (TNF-alpha) release and nitrite production measured at 72 h. Inclusion of 200 microM NG-monomethyl-L-arginine, a competitive inhibitor of the L-arginine-dependent pathway for the synthesis of inorganic nitrogen oxides, inhibited the killing, as did the addition of neutralizing anti-TNF-alpha antibody. These results are consistent with previous data showing an important autocrine role for TNF-alpha in enhancing production of inorganic nitrogen oxides by primed or activated macrophages. In addition, our results suggest that production of TNF-alpha and nitrites after priming or activation signals may be under a different regulatory control in mature bone marrow macrophages than in the resident peritoneal macrophage population.
研究了来自Lshr与Lshs同基因小鼠的成熟骨髓源性巨噬细胞和常驻腹膜巨噬细胞在重组γ干扰素-脂多糖（rIFN-γ-LPS）激活下杀死细胞内杜氏利什曼原虫无鞭毛体的能力。单独使用剂量高达100 U/ml的IFN-γ无法在体外激活巨噬细胞以杀死杜氏利什曼原虫无鞭毛体；LPS是必要的二次刺激物。同样，单独使用剂量高达100 ng/ml的LPS也未产生杀利什曼活性。在骨髓巨噬细胞中，随着引入的rIFN-γ-LPS剂量组合增加，观察到杀利什曼活性呈剂量依赖性增加，在任何特定剂量组合中，Lshr巨噬细胞都保持显著但不明显的优势。对于腹膜巨噬细胞，情况则相反，在rIFN-γ剂量为11至33 U/ml且LPS剂量高达10 ng/ml时，来自Lshs小鼠的巨噬细胞在杀伤方面更有效。骨髓和腹膜巨噬细胞中的杀伤程度与72小时时上清液中测得的亚硝酸盐水平密切相关，并且在4小时、24小时或72小时时测得的肿瘤坏死因子α（TNF-α）释放与72小时时测得的亚硝酸盐产生之间观察到高度显著的相关性。加入200 μM NG-单甲基-L-精氨酸（一种L-精氨酸依赖性无机氮氧化物合成途径的竞争性抑制剂）可抑制杀伤，加入中和抗TNF-α抗体也有同样效果。这些结果与先前的数据一致，表明TNF-α在增强致敏或活化巨噬细胞产生无机氮氧化物方面具有重要的自分泌作用。此外，我们的结果表明，在成熟骨髓巨噬细胞中，致敏或激活信号后TNF-α和亚硝酸盐的产生可能受到与常驻腹膜巨噬细胞群体不同的调控。
