Panuska J R, Fukui K, Parker C W
Howard Hughes Medical Institute Laboratory, Washington University School of Medicine, St. Louis, MO 63110.
Biochem J. 1988 Jan 15;249(2):501-11. doi: 10.1042/bj2490501.
A monocyte-rich preparation from the adherent cell fraction of human peripheral blood leukocytes was incubated for 1-8 h with [35S]methionine or [3H]leucine in the presence and absence of bacterial lipopolysaccharide (LPS). The macromolecules released into the supernatant were analysed by two-dimensional gel electrophoresis and radioautography. A complex labelling pattern involving at least 20 easily demonstrable and apparently distinct products with a broad range of molecular masses and isoelectric points was observed. LPS or LPS plus actinomycin in combination markedly stimulated the labelling and release of at least twelve different macromolecules ranging in apparent Mr from 12,000 to 46,000. Studies with monocytes that had been additionally purified by centrifugal elutriation and with the monocyte-like human cell line U-937 indicated that monocytes rather than contaminating cells were the source of these products. The majority of the secreted products were unique and did not cross-react with antibodies to interleukin 1 or tumour necrosis factor. The high resolving capacity of two-dimensional gel electrophoresis may be useful to define further the diverse biological activities and potential monokines released from monocytes at various stages of their differentiation and activation.
将人外周血白细胞贴壁细胞部分富含单核细胞的制剂，在有和没有细菌脂多糖（LPS）存在的情况下，与[35S]甲硫氨酸或[3H]亮氨酸一起孵育1 - 8小时。通过二维凝胶电泳和放射自显影分析释放到上清液中的大分子。观察到一种复杂的标记模式，涉及至少20种易于证明且明显不同的产物，其分子量和等电点范围广泛。LPS或LPS与放线菌素联合使用显著刺激了至少12种不同大分子的标记和释放，其表观分子量在12,000至46,000之间。对通过离心淘析进一步纯化的单核细胞以及单核细胞样人细胞系U - 937的研究表明，这些产物的来源是单核细胞而非污染细胞。大多数分泌产物是独特的，并且不与白细胞介素1或肿瘤坏死因子的抗体发生交叉反应。二维凝胶电泳的高分辨率能力可能有助于进一步确定单核细胞在其分化和激活的各个阶段释放的多种生物活性和潜在单核因子。
