CYTOKINE-INDUCED AND LPS-INDUCED SYNTHESIS OF INTERLEUKIN-8 FROM HUMAN MESANGIAL CELLS

被引:69
|
作者
KUSNER, DJ
LUEBBERS, EL
NOWINSKI, RJ
KONIECZKOWSKI, M
KING, CH
SEDOR, JR
机构
[1] UNIV HOSP CLEVELAND, DEPT MED, DIV NEPHROL, 2074 ABINGTON RD, CLEVELAND, OH 44106 USA
[2] UNIV HOSP CLEVELAND, DIV GEOG MED, CLEVELAND, OH 44106 USA
[3] CASE WESTERN RESERVE UNIV, SCH MED, DEPT MED, CLEVELAND, OH 44106 USA
关键词
D O I
10.1038/ki.1991.157
中图分类号
R5 [内科学]; R69 [泌尿科学(泌尿生殖系疾病)];
学科分类号
1002 ; 100201 ;
摘要
The cytokine neutrophil-activating peptide-1/interleukin-8 (NAP/IL-8) activates neutrophils (PMN) and elicits selective diapedesis of PMN into the extracellular space. The glomerular mesangial cell (MC) is a specialized pericyte that controls glomerular filtration and synthesizes and responds to a variety of cytokines. Because of its location within the glomerulus, the MC is in a pivotal position to orchestrate events underlying immune injury. Since immune-injured glomeruli have been shown to produce NAP/IL-8 activity in vitro, we assessed whether lipopolysaccharide (LPS)- or cytokine-activated MC might be a source of this activity. Pure human MC, devoid of monocyte/macrophage and fibroblast contamination, were grown by explant from collagenase-treated glomeruli. Human recombinant interleukin-1-alpha (IL-1-alpha, 20 ng/ml), IL-1-beta (50 ng/ml), tumor necrosis factor-alpha (TNF, 100 ng/ml) and lipopolysaccharide (LPS, 10-mu-g/ml) stimulated release of a neutrophil chemotactic factor from cultured MC. Both concentrated (fivefold) and unconcentrated MC supernatants stimulated directed neutrophil migration under agarose at a level similar to that of the bacterial chemotactic factor, FMLP. In contrast, unstimulated MC-conditioned media and IL-1-alpha, IL-1-beta, TNF and LPS in medium alone did not directly induce PMN migration. Molecular sizing studies using sequential membrane ultrafiltration identified significant TNF-stimulated, MC-derived chemotactic activity in the 3000 to 10000 kD fraction. An anti-NAP/IL-8 monoclonal antibody, 46E5, significantly inhibited PMN chemotaxis stimulated by TNF-stimulated, MC-conditioned media in a dose-dependent manner. S1 nuclease analyses of a total RNA extracted from TNF-stimulated MC showed a dose- and time-dependent increase in NAP/IL-8 mRNA transcripts and demonstrated that a chemotaxin, produced by cytokine- and LPS-stimulated MC, is highly homologous to NAP/IL-8. Northern analysis of TNF-stimulated MC RNA identified NAP-1/IL-8 transcripts in MC which comigrated with the 1.8 kb message in LPS-stimulated human monocytes. Using [S-35]labelled oligonucleotide probes, NAP/IL-8 mRNA transcripts were demonstrated in the cytoplasm of IL-1 and TNF-stimulated, but not control, mesangial cells. Taken together, these data demonstrate the human MC synthesize and release NAP/IL-8, a potent chemotactic peptide for PMN that may contribute to the structural and functional derangements which occur in glomerular inflammation.
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收藏
页码:1240 / 1248
页数:9
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