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脂多糖(LPS)刺激的人血单核白细胞产生的中性粒细胞趋化因子:部分特性及与白细胞介素1(IL-1)的分离

Neutrophil chemotactic factor produced by lipopolysaccharide (LPS)-stimulated human blood mononuclear leukocytes: partial characterization and separation from interleukin 1 (IL 1).

作者信息

Yoshimura T, Matsushima K, Oppenheim J J, Leonard E J

出版信息

J Immunol. 1987 Aug 1;139(3):788-93.

PMID:3298433
Abstract

LPS stimulated human blood mononuclear leukocytes to produce a chemotactic factor for human neutrophils. The effect of LPS was dose-dependent; 10 micrograms/ml was optimal for production of chemotactic factor. Chemotactic activity was detected 3 hr after LPS stimulation, and reached its peak at 12 hr. No activity was detected in culture supernatants of unstimulated cells, provided LPS-free media were selected. Isoelectric point of the factor, determined by chromatofocusing, was approximately 8 to 8.5. Molecular weight was approximately 10 kilodaltons by Sephacryl S-200 gel filtration or by HPLC gel filtration on TSK-2000 and -3000 columns in succession. The gel filtration fractions were also assayed for IL 1 activity. The elution position of IL 1 activity corresponded to a m.w. of 18. There was no chemotactic activity in the IL 1 activity peak. Furthermore, highly purified natural Il 1 alpha and -beta and recombinant Il 1 alpha and -beta did not exhibit chemotactic activity for neutrophils in our assay. Among mononuclear leukocytes, the monocyte was the principal producer of neutrophil chemotactic factor. These results suggest that a chemotactic factor for neutrophils, different from IL 1, is produced by LPS-stimulated blood monocytes.

摘要

脂多糖刺激人血单核白细胞产生一种针对人中性粒细胞的趋化因子。脂多糖的作用呈剂量依赖性;10微克/毫升最有利于趋化因子的产生。脂多糖刺激3小时后可检测到趋化活性,并在12小时达到峰值。如果选用无脂多糖的培养基,未刺激细胞的培养上清液中未检测到活性。通过色谱聚焦法测定,该因子的等电点约为8至8.5。通过Sephacryl S - 200凝胶过滤或在TSK - 2000和 - 3000柱上连续进行HPLC凝胶过滤,分子量约为10千道尔顿。还对凝胶过滤级分进行了白细胞介素1(IL 1)活性测定。IL 1活性的洗脱位置对应于分子量18。在IL 1活性峰中没有趋化活性。此外,在我们的检测中,高度纯化的天然IL 1α和 -β以及重组IL 1α和 -β对中性粒细胞均未表现出趋化活性。在单核白细胞中,单核细胞是中性粒细胞趋化因子的主要产生者。这些结果表明,脂多糖刺激的血液单核细胞产生了一种不同于IL 1的中性粒细胞趋化因子。

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