title: Monokine-induced neutrophil chemotactic factor gene expression in human fibroblasts.
abstract: Although fibroblasts are important in providing a structural framework for most tissues, they also appear to be active participants in the inflammatory process via the production of specific mediators. The production of inflammatory mediators by fibroblasts is especially important in relation to their strategic location within connective tissue as they may act as a cellular communication bridge between the interstitium and vasculature. In this paper, we demonstrate that fibroblasts may participate in these inflammatory reactions by the production of a neutrophil chemotactic factor (NCF) with characteristics similar to a recently isolated and cloned monocyte-derived NCF. Either tumor necrosis factor-alpha-, interleukin-1 alpha-, or interleukin-1 beta-stimulated fibroblasts showed both a time- and dose-dependent increase in steady-state levels of NCF mRNA and secretion of chemotactic activity. In contrast, lipopolysaccharide and interleukin-6 failed to induce fibroblast-derived NCF. The expression of fibroblast-derived NCF mRNA was first detectable by 30 min poststimulation, whereas chemotactic activity was significantly observed 3-4 h postchallenge. Heat-inactivated monokine (100 degrees C) failed to induce NCF mRNA expression, suggesting that only the active proteins are capable of inducing NCF. Gel filtration analysis using high pressure liquid chromatography indicated peak chemotactic activity with an approximate molecular mass of 8000 daltons. This peak of NCF activity was found to be relatively stable to both heat and trypsin inactivation. Specificity of the fibroblast-derived neutrophil chemotactic activity was demonstrated with inhibition of chemotaxis by the addition of neutralizing antibody directed against recombinant human neutrophil chemotactic factor. These data provide evidence that monokine-treated fibroblasts can synthesize a potent chemotactic agent with molecular and physicochemical characteristics similar to monocyte-derived NCF and that this factor may contribute to neutrophil-mediated disease processes.
authors.affiliation: Department of Pathology, University of Michigan Medical School, Ann Arbor 48109-0602.
authors.forename: R M
authors.initials: RM
authors.lastname: Strieter
authors.forename: S H
authors.initials: SH
authors.lastname: Phan
authors.forename: H J
authors.initials: HJ
authors.lastname: Showell
authors.forename: D G
authors.initials: DG
authors.lastname: Remick
authors.forename: J P
authors.initials: JP
authors.lastname: Lynch
authors.forename: M
authors.initials: M
authors.lastname: Genord
authors.forename: C
authors.initials: C
authors.lastname: Raiford
authors.forename: M
authors.initials: M
authors.lastname: Eskandari
authors.forename: R M
authors.initials: RM
authors.lastname: Marks
authors.forename: S L
authors.initials: SL
authors.lastname: Kunkel
mesh_terms: D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D002478:Cells, Cultured; D002630:Chemotactic Factors / Q000235:genetics*; D015342:DNA Probes; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005796:Genes / Q000187:drug effects*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D015850:Interleukin-6; D016209:Interleukin-8; D007378:Interleukins / Q000494:pharmacology*; D007700:Kinetics; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism*; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics*; D011994:Recombinant Proteins / Q000494:pharmacology; D012867:Skin / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D002352:Carrier Proteins; D002630:Chemotactic Factors; D015342:DNA Probes; D007375:Interleukin-1; D015850:Interleukin-6; D016209:Interleukin-8; D007378:Interleukins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha