Characterization of a human eosinophil proteoglycan, and augmentation of its biosynthesis and size by interleukin 3, interleukin 5, and granulocyte/macrophage colony stimulating factor.
Characterization of a human eosinophil proteoglycan, and augmentation of its biosynthesis and size by interleukin 3, interleukin 5, and granulocyte/macrophage colony stimulating factor.
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DOI：
10.1016/s0021-9258(18)68329-2
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发表时间：
1988-09
期刊：
影响因子：
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通讯作者：
M. E. Rothenberg;J L Pomerantz;W. F. Owen;S. Avraham;R. Soberman;K F Austen;R. Stevens
中科院分区：
文献类型：
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作者：
M. E. Rothenberg;J L Pomerantz;W. F. Owen;S. Avraham;R. Soberman;K F Austen;R. Stevens
在不存在或存在重组人白细胞介素（IL）3、小鼠IL 5或重组人粒细胞/巨噬细胞集落刺激因子（GM-CSF）的情况下，将人嗜酸性粒细胞在富集培养基中培养长达7天，然后用[35 S]硫酸盐进行放射性标记，以表征其细胞相关蛋白聚糖。未暴露于任何这些细胞因子的新鲜分离的嗜酸性粒细胞合成Mr约80，000的链霉蛋白酶抗性35 S标记的蛋白聚糖，其含有Mr约80，000的糖胺聚糖。用编码早幼粒细胞白血病HL-60细胞蛋白聚糖肽核心的cDNA对总嗜酸性粒细胞RNA进行RNA印迹分析，发现嗜酸性粒细胞中编码类似分子的mRNA与HL-60细胞中的相似，约为1.3个组氨酸。当嗜酸性粒细胞在10 pM IL 3、1 pM IL 5或10 pM GM-CSF存在下培养1天或更长时间时，[35 S]硫酸盐掺入率增加约2倍，细胞合成Mr约300，000的链霉蛋白酶抗性35 S标记的蛋白聚糖，其中含有Mr约30，000的35 S标记的糖胺聚糖。约93%的35 S-标记的糖胺聚糖结合到由noncytokine和甘氨酸处理的嗜酸性粒细胞合成的蛋白聚糖，对软骨素酶ABC降解敏感。如通过高效液相色谱法所评估的，这些软骨素酶ABC产生的35 S标记的二糖中的6-16%是衍生自硫酸软骨素E的二硫酸化二糖;其余的是衍生自硫酸软骨素A的单硫酸化二糖。利用GM-CSF作为细胞因子的模型，证明了GM-CSF处理的细胞比非精氨酸处理的细胞在β-D-木糖苷上合成更大的糖胺聚糖。因此，IL 3、IL 5和GM-CSF诱导人嗜酸性粒细胞通过增加新合成的蛋白聚糖及其单独的硫酸软骨素链的大小来增加蛋白聚糖的生物合成。
Human eosinophils were cultured for up to 7 days in enriched medium in the absence or presence of recombinant human interleukin (IL) 3, mouse IL 5, or recombinant human granulocyte/macrophage colony stimulating factor (GM-CSF) and then were radiolabeled with [35S]sulfate to characterize their cell-associated proteoglycans. Freshly isolated eosinophils that were not exposed to any of these cytokines synthesized Mr approximately 80,000 Pronase-resistant 35S-labeled proteoglycans which contained Mr approximately 80,000 glycosaminoglycans. RNA blot analysis of total eosinophil RNA, probed with a cDNA that encodes a proteoglycan peptide core of the promyelocytic leukemia HL-60 cell, revealed that the mRNA which encodes the analogous molecule in eosinophils was approximately 1.3 kilobases, like that in HL-60 cells. When eosinophils were cultured for 1 day or longer in the presence of 10 pM IL 3, 1 pM IL 5, or 10 pM GM-CSF, the rates of [35S]sulfate incorporation were increased approximately 2-fold, and the cells synthesized Mr approximately 300,000 Pronase-resistant 35S-labeled proteoglycans which contained Mr approximately 30,000 35S-labeled glycosaminoglycans. Approximately 93% of the 35S-labeled glycosaminoglycans bound to the proteoglycans synthesized by noncytokine- and cytokine-treated eosinophils were susceptible to degradation by chondroitinase ABC. As assessed by high performance liquid chromatography, 6-16% of these chondroitinase ABC-generated 35S-labeled disaccharides were disulfated disaccharides derived from chondroitin sulfate E; the remainder were monosulfated disaccharides derived from chondroitin sulfate A. Utilizing GM-CSF as a model of the cytokines, it was demonstrated that the GM-CSF-treated cells synthesized larger glycosaminoglycans onto beta-D-xyloside than the noncytokine-treated cells. Thus, IL 3, IL 5, and GM-CSF induce human eosinophils to augment proteoglycan biosynthesis by increasing the size of the newly synthesized proteoglycans and their individual chondroitin sulfate chains.