The differential production of three forms of IL-1 receptor antagonist by human neutrophils and monocytes.
The differential production of three forms of IL-1 receptor antagonist by human neutrophils and monocytes.
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DOI：
10.4049/jimmunol.161.4.2004
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发表时间：
1998-08
影响因子：
4.4
通讯作者：
M. Malyak;Michael F. Smith;Ashley A. Abel;K. Hance;W. Arend
中科院分区：
文献类型：
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作者：
M. Malyak;Michael F. Smith;Ashley A. Abel;K. Hance;W. Arend
IL-1 R拮抗剂（IL-1 Ra）以三种充分表征的亚型存在。17 kDa分泌型IL-1 Ra（sIL-1 Ra）和18 kDa胞内型IL-1 Ra（icIL-1 Ra I）是由同一IL-1 Ra基因的交替转录产生的。最近描述的16-kDa细胞内IL-1 Ra（icIL-1 Ra II）是由sIL-1 Ra mRNA的交替翻译起始形成的。使用RT-PCR、ELISA和Western印迹分析检测LPS刺激的人中性粒细胞和PBMC中IL-1 Ra亚型的转录和翻译。LPS刺激中性粒细胞导致sIL-1 Ra mRNA水平升高1小时，而icIL-1 RaI mRNA在22小时的培养中仍然检测不到。在LPS刺激的中性粒细胞培养3 h后，观察到细胞外糖基化sIL-1 Ra蛋白和细胞内icIL-1 RaII;免疫印迹法未检测到icIL-1 RaI蛋白。LPS刺激PBMC 1h后sIL-1 Ra mRNA水平升高，8h后可检测到icIL-1 RaI mRNA。LPS刺激的PBMC表现出细胞外糖基化的sIL-1 Ra蛋白和细胞内的icIL-1 RaII的刺激3小时内，而检测到的icIL-1 RaI蛋白延迟到15小时的培养。亚细胞定位实验表明，icIL-1 RaI和icIL-1 RaII主要存在于细胞质区室中，正如预期的那样，它们缺乏信号肽。这些结果表明，尽管LPS刺激的中性粒细胞和PBMC都合成sIL-1 Ra和icIL-1 Ra II，但只有PBMC转录和翻译icIL-1 Ra I。此外，sIL-Ra转录和翻译（以及icIL-1 RaII的翻译）是早期事件，而PBMC中的icIL-1 RaI转录延迟。
IL-1R antagonist (IL-1Ra) exists as three well-characterized isoforms. The 17-kDa secretory IL-1Ra (sIL-1Ra) and 18-kDa intracellular IL-1Ra (icIL-1RaI) arise by alternative transcription of the same IL-1Ra gene. The recently described 16-kDa intracellular IL-1Ra (icIL-1RaII) is formed by alternative translation initiation of sIL-1Ra mRNA. Transcription and translation of IL-1Ra isoforms were examined in LPS-stimulated human neutrophils and PBMC using RT-PCR, ELISA, and Western blot analysis. LPS stimulation of neutrophils resulted in elevated sIL-1Ra mRNA levels by 1 h, whereas icIL-1RaI mRNA remained undetectable through 22 h of culture. Extracellular glycosylated sIL-1Ra protein and intracellular icIL-1RaII were observed in LPS-stimulated neutrophils by 3 h of culture; no icIL-1RaI protein was detected by immunoblot. LPS stimulation of PBMC resulted in elevated sIL-1Ra mRNA levels by 1 h and detectable icIL-1RaI mRNA at 8 h of culture. LPS-stimulated PBMC demonstrated extracellular glycosylated sIL-1Ra protein and intracellular icIL-1RaII within 3 h of stimulation, whereas detection of icIL-1RaI protein was delayed until 15 h of culture. Subcellular localization experiments established that both icIL-1RaI and icIL-1RaII were present primarily within the cytoplasmic compartment, as expected by their lack of a signal peptide. These results demonstrate that although both LPS-stimulated neutrophils and PBMC synthesize sIL-1Ra and icIL-1RaII, only PBMC transcribe and translate icIL-1RaI. Furthermore, sIL-Ra transcription and translation (and translation of icIL-1RaII) are early events, whereas icIL-1RaI transcription in PBMC is delayed.