Potent synergistic effect of IL-3 and TNF on matrix metalloproteinase 9 generation by human eosinophils

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• 作者：Elizabeth A.B. Kelly¹ ; ^{eak@medicine.wisc.edu} ; Lin Ying Liu¹ ; ^{lliu6@bidmc.harvard.edu} ; Stephane Esnault ^{sesnault@medicine.wisc.edu} ; Beatriz Helena Quinchia Johnson ^{beatrizq@msn.com} ; Nizar N. Jarjour ^{nnj@medicine.wisc.edu}
• 关键词：Ab ; antibody ; 尾c ; common beta chain ; Ct ; comparative cycle threshold ; ECM ; extracellular matrix ; ERK ; extracellular signal-regulated protein kinase ; GUS ; glucuronidase ; MAP ; mitogen-activated protein kinase ; MEK ; mitogen-activated protein kinase kinase
• 刊名：Cytokine
• 出版年：2012
• 期刊代码：97_10434666
• 类别：imm
• 出版时间：May, 2012
• 卷：58
• 期：2
• 页码：199-206
• 文件大小：779 K

摘要

TNF (designated as TNF-伪 under previous nomenclature) is the preeminent activator of MMP-9 generation from a variety of cells including eosinophils. We have previously established that TNF strongly synergizes with IFN-纬 and IL-4 for eosinophil synthesis of Th1- and Th2-type chemokines respectively. Thus, we sought to determine if TNF-induced synthesis of MMP-9 would be enhanced by the presence of Th1, Th2, or the eosinophil-associated common beta chain (尾c) cytokines. Human blood eosinophils were cultured with TNF alone or in combination with either IFN-纬, IL-4, IL-3, IL-5, or GM-CSF. Concentrations and activities of MMP-9 in eosinophil culture supernates were measured by ELISA and gelatin zymography, mRNA transcription and stabilization by quantitative real-time PCR, and signaling events by immunoblotting and intracellular flow cytometric analysis. Individually, TNF, GM-CSF, or IL-3, but not IL-4 or IFN-纬, induced relatively small (<0.2 ng/ml) but statistically significant quantities of MMP-9. Remarkable synergistic synthesis of MMP-9 (ng/ml levels) occurred in response to TNF plus IL-3, GM-CSF or IL-5, in the order of IL-3 > GM-CSF > IL-5. Zymography revealed that eosinophils release MMP-9 in its pro-form. Eosinophil stimulation with the combination of IL-3 plus TNF led to increased steady-state levels of MMP-9 mRNA, prolonged mRNA stabilization, and enhanced activation of ERK1/2 phosphorylation. Inhibition of NF-魏B, MEK kinase, or p38 MAP kinase, but not JNK signaling pathways, diminished IL-3/TNF-induced MMP-9 mRNA and protein production. Thus, the synergistic regulation of eosinophil MMP-9 by IL-3 plus TNF likely involves cooperative interaction of multiple transcription factors downstream from ERK, p38, and NF-魏B activation as well as post-transcriptional regulation of MMP-9 mRNA stabilization. Our data indicate that within microenvironments rich in 尾c-family cytokines and TNF, eosinophils are an important source of proMMP-9 and highlight a previously unrecognized role for synergistic interaction between TNF and 尾c-family cytokines, particularly IL-3, for proMMP-9 synthesis.