Production of Proinflammatory Mediators by Indoor Air Bacteria and Fungal Spores in Mouse and Human Cell Lines. (Research)

By Huttunen, Kati; Hyvarinen, Anne et al. | Environmental Health Perspectives, January 2003 | Go to article overview
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Production of Proinflammatory Mediators by Indoor Air Bacteria and Fungal Spores in Mouse and Human Cell Lines. (Research)


Huttunen, Kati, Hyvarinen, Anne, Nevalainen, Aino, Komulainen, Hannu, Hirvonen, Maija-Riitta, Environmental Health Perspectives


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.sup.5], [10.sup.6], and [10.sup.7] microbes/mL. We studied time dependency by terminating the exposure to [10.sup.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 1[beta] (TNF-[alpha], IL-6, IL-1[beta], 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-1[beta] 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. Key words: bacteria, cytokine production, fungi, inflammation, mold. Environ Health Perspect 111:85-92 (2003). [Online 4 December 2002] doi:10.1289/ehp.5478 available via http://dx.doi.org/

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Exposure to microbes is recognized as a potential cause of inflammation-related health problems among occupants of moldy buildings (Husman 1996; Peat et al. 1998). Many different microbes, including a variety of fungi and bacteria, thrive in the conditions offered by moist building materials, which contain both the nutrients and the moisture needed for microbial growth (Hyvarinen et al. 2002; Murtoniemi et al. 2001; Nikulin et al. 1994). The current understanding is that microbial growth affects indoor air quality, and occupants are exposed to biological pollutants, which may lead to adverse health effects. Currently it is not known which components of the microbial flora are most harmful to occupants of moldy buildings. Comparisons of the different microbes present in such environments are required to evaluate the potential health effects of a given microbe. Our working hypothesis is that bacteria isolated from moldy buildings are also important in causing inflammatory and cytotoxic responses.

Indoor bacterial flora include both gram-positive and gram-negative strains, which grow in moist building materials (Andersson et al. 1997; Hyvarinen et al. 2002). The gram-positive sporulating bacteria Streptomyces spp. have been frequently isolated from water-damaged buildings, but they are not part of the normal microbial flora of indoor air (Nevalainen et al. 1991). The presence of Streptomyces is also an indicator of moisture damage in buildings (Samson et al. 1994). We demonstrated previously that the spores of streptomycetes originating from moldy buildings can evoke intense production of inflammatory mediators both in mouse and human cell lines (Hirvonen et al. 1997; Jussila et al. 1999) in vitro and in mouse lungs in vivo (Jussila et al. 2001). Thus, the correlation found between in vitro and in vivo data strongly supports the view that streptomycetes play a role in the cascade of events leading to adverse health effects in occupants of moldy buildings.

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Production of Proinflammatory Mediators by Indoor Air Bacteria and Fungal Spores in Mouse and Human Cell Lines. (Research)
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