title: Expression of TNF and the necessity of TNF receptors in bleomycin-induced lung injury in mice.
abstract: Bleomycin (BLM) induction of lung fibrosis in mice is an established model to study the mechanism of pulmonary fibrosis. Cytokine secretion has been implicated as a fundamental component of the lung fibrotic process observed in response to BLM. Among the cytokines implicated in lung fibrosis, Tumor necrosis factor (TNF) alpha has been considered to play a fundamental role. In the present study, we characterized the cellular sources of TNF during BLM-induced lung injury and examined the importance of TNF receptors in this process. To characterize the expression of TNF, we utilized two strains of mice, one sensitive (C57BL/6) and one resistant (BALB/c) to BLM-induced lung injury. Mice received BLM (120 mg/kg total) or saline, as control, by multiple subcutaneous injections. BLM induced the development of inflammation in subpleural areas only in the lungs of BLM-sensitive mice. These subpleural areas were characterized by infiltration of CD68-positive macrophages and increased collagen deposition. BLM enhanced the expression of TNF mRNA in BLM-sensitive, but not in BLM-resistant, mice. In situ hybridization studies localized the expression of TNF in the areas of BLM-induced inflammation in 6% and 27% of macrophages at 14 and 21 days post BLM treatment. In addition to TNF, BLM exposure resulted in the upregulated expression of transforming growth factor (TGF)-beta 1, but not interleukin (IL)-1, mRNA in the lungs of both murine strains at 14 and 21 days. This upregulated expression of TGF-beta 1 mRNA was greater in the lungs of BLM-sensitive mice. In separate experiments, double TNF receptor knockout mice were exposed to BLM. These animals demonstrated an increased expression of TNF, but not TGF-beta 1, mRNA in response to BLM and did not exhibit histologic evidence of lung injury following BLM exposure. In summary, the upregulation of TNF mRNA in macrophages correlated with the appearance of inflammation following BLM exposure and was limited to the BLM-sensitive strain. Furthermore, in addition to the release of the TNF ligand, it appears that the presence of TNF receptors is necessary for the development of BLM-induced lung injury, and signaling through these receptors may contribute to the regulation of the TGF-beta 1 mRNA expression observed in response to bleomycin. These results provide further support for a role of macrophages and TNF in the induction of lung inflammation.
authors.affiliation: Department of Medicine, Tulane University Medical Center, New Orleans, Louisiana 70112-2699, USA. lortiz@mailhost.tcs.tulane.edu
authors.forename: L A
authors.initials: LA
authors.lastname: Ortiz
authors.forename: J
authors.initials: J
authors.lastname: Lasky
authors.forename: R F
authors.initials: RF
authors.lastname: Hamilton
authors.forename: A
authors.initials: A
authors.lastname: Holian
authors.forename: G W
authors.initials: GW
authors.lastname: Hoyle
authors.forename: W
authors.initials: W
authors.lastname: Banks
authors.forename: J J
authors.initials: JJ
authors.lastname: Peschon
authors.forename: A R
authors.initials: AR
authors.lastname: Brody
authors.forename: G
authors.initials: G
authors.lastname: Lungarella
authors.forename: M
authors.initials: M
authors.lastname: Friedman
mesh_terms: D000818:Animals; D001761:Bleomycin / Q000633:toxicity; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D005838:Genotype; D006909:Hydroxyproline / Q000378:metabolism; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D007375:Interleukin-1 / Q000378:metabolism; D008168:Lung / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D011658:Pulmonary Fibrosis / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000096:biosynthesis; D018124:Receptors, Tumor Necrosis Factor / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D013047:Specific Pathogen-Free Organisms; D016212:Transforming Growth Factor beta / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D007375:Interleukin-1; D012333:RNA, Messenger; D018124:Receptors, Tumor Necrosis Factor; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D001761:Bleomycin; D006909:Hydroxyproline