Modulation of multiple neutrophil functions by preparative methods or trace concentrations of bacterial lipopolysaccharide.
Modulation of multiple neutrophil functions by preparative methods or trace concentrations of bacterial lipopolysaccharide.
复制标题
DOI：
--
复制
发表时间：
1985-04
期刊：
影响因子：
--
通讯作者：
C. Haslett;L. Guthrie;Marguerite M. Kopaniak;B. Richard;Johnston;P. Henson
中科院分区：
文献类型：
--
作者：
C. Haslett;L. Guthrie;Marguerite M. Kopaniak;B. Richard;Johnston;P. Henson
通过四种方法从外周血中分离人嗜中性粒细胞：1）Ficoll-Hypaque梯度和红细胞裂解，2）血浆-Percoll梯度，3）产生85%嗜中性粒细胞的“无脂多糖（LPS）”方法，和4）通过血浆-Percoll梯度离心方法3制备的细胞以产生纯嗜中性粒细胞。与其他方法相比，Ficoll-Hypaque方法的使用导致细胞形状的自发变化，增强了甲酰甲硫氨酰亮氨酰苯丙氨酸（FMLP）刺激的超氧阴离子释放，增加了随后FMLP刺激后溶酶体酶的释放，并降低了趋化反应。这些效应不是由于NH 4Cl溶解红细胞所致，而是通过将“无LPS”方法制备的中性粒细胞暴露于10-100 ng/ml LPS或使用血浆Percoll梯度来再现。中性粒细胞的形状变化和刺激O-2的产生是这些影响的特别敏感的标志。微量浓度的LPS在调节中性粒细胞功能中的作用可能与内毒素血症的病理生理学及其导致的组织损伤有关。
Human neutrophils were isolated from peripheral blood by four methods: 1) Ficoll-Hypaque gradients and erythrocyte lysis, 2) plasma-Percoll gradients, 3) a "lipopolysaccharide (LPS)-free" method yielding 85% neutrophils, and 4) by centrifugation of cells prepared by Method 3 through a plasma-Percoll gradient to produce pure neutrophils. The use of the Ficoll-Hypaque method resulted in spontaneous change of cell shape, enhanced formyl-methionyl-leucyl-phenylalanine (FMLP)-stimulated release of superoxide anion, increased release of lysosomal enzymes upon subsequent FMLP stimulation, and reduced chemotactic responsiveness, by comparison with the other methods. These effects were not due to erythrocyte lysis by NH4C1 but were reproduced by exposure of neutrophils prepared by the "LPS-free" method or the use of plasma-Percoll gradients to 10-100 ng/ml LPS. Neutrophil change of shape and stimulated O-2 production were particularly sensitive markers of these effects. The effects of trace concentrations of LPS in the modulation of neutrophil function may have relevance to the pathophysiology of endotoxemia and its resultant tissue injury.