title: In vitro acute effects of tobacco smoke on tumor necrosis factor alpha and interleukin-6 production by alveolar macrophages.
abstract: Tobacco smoke is a usual form of oxidant aggression present in the domestic environment. In the present study, the in vitro acute effects of a 2-cigarette smoke gas phase were evaluated on cell viability and cytokine secretion by alveolar macrophages (AM) from guinea pigs and human healthy subjects. Cell injury was estimated immediately after smoke exposure by evaluation of ATP cell content (measured by bioluminescence) and lactic dehydrogenase (LDH) release in the culture medium. LDH release was also measured when the interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF) activities were evaluated. No cytotoxic effect was found: The ATP cell content of both guinea pig AM and human AM did not significantly change after tobacco smoke exposure. Similarly, the LDH release in the culture medium was unchanged both immediately after tobacco smoke exposure and at the time of the cytokine evaluation (18-20 h later) compared to cells cultured in the air. The total protein synthesis by the guinea pig AM evaluated by 35S-L-methionine labeling was unaffected by tobacco smoke exposure. The production of IL-6 and TNF activities was evaluated 18-20 h after smoke exposure. The IL-6 activity was measured by the proliferation test of 7TD1 hybridoma cell line; the TNF activity was evaluated by the L929 mouse fibroblast cytotoxic test and by an immunoradiometric assay (for human AM). A 2-cigarette smoke exposure decreased both activities significantly. The exposure of the guinea pig AM reduced IL-6 activity by 24.3 +/- 6.7%, 42.4 +/- 7.8%, and 39.7 +/- 9.6% and TNF activity by 33.8 +/- 10.4%, 35.1 +/- 10.7%, and 38.8 +/- 9.9% (respectively unstimulated cells and AM activated by 0.1 and 10 micrograms LPS/mL). The decrease in monokine production by the human AM was, respectively, 57.8 +/- 8.8%, 59.7 +/- 11.4%, and 49.9 +/- 10.5% of IL-6 activity and 37.4 +/- 14.6%, 17.6 +/- 9.6%, and 37.2 +/- 6.3% of TNF activity. The possible release of cytokine inhibitors was also investigated. The inhibitory activity against recombinant TNF and IL-6 was evaluated in culture medium from unstimulated AM exposed to tobacco smoke and did not significantly differ from that of AM exposed to air, demonstrating that the decrease of monokine levels could not be explained by the release of inhibitory factors.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Laboratoire de Pathologie Respiratoire Expérimentale et de Pollution, Atmosphérique, Institut Pasteur, Lille, France.
authors.forename: V
authors.initials: V
authors.lastname: Dubar
authors.forename: P
authors.initials: P
authors.lastname: Gosset
authors.forename: C
authors.initials: C
authors.lastname: Aerts
authors.forename: C
authors.initials: C
authors.lastname: Voisin
authors.forename: B
authors.initials: B
authors.lastname: Wallaert
authors.forename: A B
authors.initials: AB
authors.lastname: Tonnel
mesh_terms: D000818:Animals; D002460:Cell Line; D006168:Guinea Pigs; D006801:Humans; D006825:Hybridomas / Q000473:pathology; D015850:Interleukin-6 / Q000096:biosynthesis*; D016676:Macrophages, Alveolar / Q000378:metabolism*; D012907:Smoking / Q000009:adverse effects; D014028:Tobacco Smoke Pollution / Q000009:adverse effects*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
chemical_list: D015850:Interleukin-6; D014028:Tobacco Smoke Pollution; D014409:Tumor Necrosis Factor-alpha