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Environ Health Perspect. 2003 Jan; 111(1): 85–92.
PMCID: PMC1241310
Research Article

Production of proinflammatory mediators by indoor air bacteria and fungal spores in mouse and human cell lines.


We compared the inflammatory and cytotoxic responses caused by household mold and bacteria in human and mouse cell lines. We studied the fungi Aspergillus versicolor, Penicillium spinulosum, and Stachybotrys chartarum and the bacteria Bacillus cereus, Pseudomonas fluorescens, and Streptomyces californicus for their cytotoxicity and ability to stimulate the production of inflammatory mediators in mouse RAW264.7 and human 28SC macrophage cell lines and in the human A549 lung epithelial cell line in 24-hr exposure to 10(5), 10(6), and 10(7) microbes/mL. We studied time dependency by terminating the exposure to 10(6) microbes/mL after 3, 6, 12, 24, and 48 hr. We analyzed production of the cytokines tumor necrosis factor-alpha and interleukins 6 and 1ss (TNF-alpha, IL-6, IL-1ss, respectively) and measured nitric oxide production using the Griess method, expression of inducible NO-synthase with Western Blot analysis, and cytotoxicity with the MTT-test. All bacteria strongly induced the production of TNF-alpha, IL-6 and, to a lesser extent, the formation of IL-1ss in mouse macrophages. Only the spores of Str. californicus induced the production of NO and IL-6 in both human and mouse cells. In contrast, exposure to fungal strains did not markedly increase the production of NO or any cytokine in the studied cell lines except for Sta. chartarum, which increased IL-6 production somewhat in human lung epithelial cells. These microbes were less cytotoxic to human cells than to mouse cells. On the basis of equivalent numbers of bacteria and spores of fungi added to cell cultures, the overall potency to stimulate the production of proinflammatory mediators decreased in the order Ps. fluorescens > Str. californicus > B. cereus > Sta. chartarum > A. versicolor > P. spinulosum. These data suggest that bacteria in water-damaged buildings should also be considered as causative agents of adverse inflammatory effects.

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Selected References

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  • Anderson AS, Wellington EM. The taxonomy of Streptomyces and related genera. Int J Syst Evol Microbiol. 2001 May;51(Pt 3):797–814. [PubMed]
  • Andersson MA, Nikulin M, Köljalg U, Andersson MC, Rainey F, Reijula K, Hintikka EL, Salkinoja-Salonen M. Bacteria, molds, and toxins in water-damaged building materials. Appl Environ Microbiol. 1997 Feb;63(2):387–393. [PMC free article] [PubMed]
  • Andersson AM, Weiss N, Rainey F, Salkinoja-Salonen MS. Dust-borne bacteria in animal sheds, schools and children's day care centres. J Appl Microbiol. 1999 Apr;86(4):622–634. [PubMed]
  • Batrakov SG, Bergelson LD. Lipids of the Streptomycettes. Structural investigation and biological interrelation a review. Chem Phys Lipids. 1978 Apr;21(1-2):1–29. [PubMed]
  • Brightbill HD, Libraty DH, Krutzik SR, Yang RB, Belisle JT, Bleharski JR, Maitland M, Norgard MV, Plevy SE, Smale ST, et al. Host defense mechanisms triggered by microbial lipoproteins through toll-like receptors. Science. 1999 Jul 30;285(5428):732–736. [PubMed]
  • de Vera ME, Shapiro RA, Nussler AK, Mudgett JS, Simmons RL, Morris SM, Jr, Billiar TR, Geller DA. Transcriptional regulation of human inducible nitric oxide synthase (NOS2) gene by cytokines: initial analysis of the human NOS2 promoter. Proc Natl Acad Sci U S A. 1996 Feb 6;93(3):1054–1059. [PMC free article] [PubMed]
  • Dörger M, Jesch NK, Rieder G, Hirvonen MR, Savolainen K, Krombach F, Messmer K. Species differences in NO formation by rat and hamster alveolar macrophages in vitro. Am J Respir Cell Mol Biol. 1997 Apr;16(4):413–420. [PubMed]
  • Flappan SM, Portnoy J, Jones P, Barnes C. Infant pulmonary hemorrhage in a suburban home with water damage and mold (Stachybotrys atra). Environ Health Perspect. 1999 Nov;107(11):927–930. [PMC free article] [PubMed]
  • Fogelmark B, Thorn J, Rylander R. Inhalation of (1-->3)-beta-D-glucan causes airway eosinophilia. Mediators Inflamm. 2001 Feb;10(1):13–19. [PMC free article] [PubMed]
  • Ganster RW, Taylor BS, Shao L, Geller DA. Complex regulation of human inducible nitric oxide synthase gene transcription by Stat 1 and NF-kappa B. Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8638–8643. [PMC free article] [PubMed]
  • Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem. 1982 Oct;126(1):131–138. [PubMed]
  • Gutierrez HH, Pitt BR, Schwarz M, Watkins SC, Lowenstein C, Caniggia I, Chumley P, Freeman BA. Pulmonary alveolar epithelial inducible NO synthase gene expression: regulation by inflammatory mediators. Am J Physiol. 1995 Mar;268(3 Pt 1):L501–L508. [PubMed]
  • Hirvonen MR, Nevalainen A, Makkonen N, Mönkkönen J, Savolainen K. Induced production of nitric oxide, tumor necrosis factor, and interleukin-6 in RAW 264.7 macrophages by streptomycetes from indoor air of moldy houses. Arch Environ Health. 1997 Nov-Dec;52(6):426–432. [PubMed]
  • Hirvonen MR, Suutari M, Ruotsalainen M, Lignell U, Nevalainen A. Effect of growth medium on potential of Streptomyces Anulatus spores to induce inflammatory responses and cytotoxicity in RAW264.7 macrophages. Inhal Toxicol. 2001 Jan;13(1):55–68. [PubMed]
  • Husman T. Health effects of indoor-air microorganisms. Scand J Work Environ Health. 1996 Feb;22(1):5–13. [PubMed]
  • Jarvis BB, Sorenson WG, Hintikka EL, Nikulin M, Zhou Y, Jiang J, Wang S, Hinkley S, Etzel RA, Dearborn D. Study of toxin production by isolates of Stachybotrys chartarum and Memnoniella echinata isolated during a study of pulmonary hemosiderosis in infants. Appl Environ Microbiol. 1998 Oct;64(10):3620–3625. [PMC free article] [PubMed]
  • Jesch NK, Dörger M, Enders G, Rieder G, Vogelmeier C, Messmer K, Krombach F. Expression of inducible nitric oxide synthase and formation of nitric oxide by alveolar macrophages: an interspecies comparison. Environ Health Perspect. 1997 Sep;105 (Suppl 5):1297–1300. [PMC free article] [PubMed]
  • Johanning E, Biagini R, Hull D, Morey P, Jarvis B, Landsbergis P. Health and immunology study following exposure to toxigenic fungi (Stachybotrys chartarum) in a water-damaged office environment. Int Arch Occup Environ Health. 1996;68(4):207–218. [PubMed]
  • Johanning E, Landsbergis P, Gareis M, Yang CS, Olmsted E. Clinical experience and results of a Sentinel Health Investigation related to indoor fungal exposure. Environ Health Perspect. 1999 Jun;107 (Suppl 3):489–494. [PMC free article] [PubMed]
  • Jussila J, Komulainen H, Huttunen K, Roponen M, Hälinen A, Hyvärinen A, Kosma VM, Pelkonen J, Hirvonen MR. Inflammatory responses in mice after intratracheal instillation of spores of Streptomyces californicus isolated from indoor air of a moldy building. Toxicol Appl Pharmacol. 2001 Feb 15;171(1):61–69. [PubMed]
  • Lowenstein CJ, Alley EW, Raval P, Snowman AM, Snyder SH, Russell SW, Murphy WJ. Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon gamma and lipopolysaccharide. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9730–9734. [PMC free article] [PubMed]
  • Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. [PubMed]
  • Murtoniemi T, Nevalainen A, Suutari M, Toivola M, Komulainen H, Hirvonen MR. Induction of cytotoxicity and production of inflammatory mediators in raw264.7 macrophages by spores grown on six different plasterboards. Inhal Toxicol. 2001 Mar;13(3):233–247. [PubMed]
  • Nikulin M, Pasanen AL, Berg S, Hintikka EL. Stachybotrys atra Growth and Toxin Production in Some Building Materials and Fodder under Different Relative Humidities. Appl Environ Microbiol. 1994 Sep;60(9):3421–3424. [PMC free article] [PubMed]
  • Nikulin M, Reijula K, Jarvis BB, Veijalainen P, Hintikka EL. Effects of intranasal exposure to spores of Stachybotrys atra in mice. Fundam Appl Toxicol. 1997 Feb;35(2):182–188. [PubMed]
  • Pasanen AL, Yli-Pietila K, Pasanen P, Kalliokoski P, Tarhanen J. Ergosterol content in various fungal species and biocontaminated building materials . Appl Environ Microbiol. 1999 Jan;65(1):138–142. [PMC free article] [PubMed]
  • Peat JK, Dickerson J, Li J. Effects of damp and mould in the home on respiratory health: a review of the literature. Allergy. 1998 Feb;53(2):120–128. [PubMed]
  • Raetz CR, Ulevitch RJ, Wright SD, Sibley CH, Ding A, Nathan CF. Gram-negative endotoxin: an extraordinary lipid with profound effects on eukaryotic signal transduction. FASEB J. 1991 Sep;5(12):2652–2660. [PubMed]
  • Rylander R, Lin RH. (1-->3)-beta-D-glucan - relationship to indoor air-related symptoms, allergy and asthma. Toxicology. 2000 Nov 2;152(1-3):47–52. [PubMed]
  • Xie QW, Whisnant R, Nathan C. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide. J Exp Med. 1993 Jun 1;177(6):1779–1784. [PMC free article] [PubMed]

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