Interactions between Streptomyces californicus and Stachybotrys chartarum can induce apoptosis and cell cycle arrest in mouse RAW264.7 macrophages.
Exposure to complex mixtures of bacteria and fungi in moisture-damaged buildings is a potential cause of inflammatory related symptoms among occupants. The present study assessed interactions between two characteristic moldy house microbes Streptomyces californicus and Stachybotrys chartarum. Differences in cytotoxic and inflammatory responses in mouse (RAW264.7) macrophages were studied after exposure to the spores of co-cultivated microbes, the mixture of separately cultivated spores, and the spores of either of these microbes cultivated alone. The RAW264.7 cells were exposed to six doses (1 x 10(4) to 3 x 10(6) spores/ml) for 24 h, and the time course of the induced responses was evaluated after 4, 8, 16, and 24 h of exposure (1 x 10(6) spores/ml). The cytotoxic potential of the spores was characterized by the MTT test, DNA content analysis, and enzyme assay for caspase-3 activity. The production of cytokines (IL-1beta, IL-6, IL-10, TNFalpha, and MIP2) was measured immunochemically and nitric oxide by the Griess method. Co-cultivation increased the ability of the spores to cause apoptosis by more than 4-fold and the proportion of RAW264.7 cells at the G2/M stage increased nearly 2-fold when compared to the response induced by the mixture of spores. In contrast, co-cultivation decreased significantly the ability of the spores to trigger the production of NO and IL-6 in RAW264.7 cells. In conclusion, these data suggest that co-culture of S. californicus and S. chartarum can result in microbial interactions that significantly potentiate the ability of the spores to cause apoptosis and cell cycle arrest in mammalian cells.
Penttinen
Piia
P
Department of Environmental Health, National Public Health Institute, FIN-70701 Kuopio, Finland. Piia.Penttinen@ktl.fi
Pelkonen
Jukka
J
Huttunen
Kati
K
Toivola
Mika
M
Hirvonen
Maija-Riitta
MR
D000818:Animals; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism / Q000382:microbiology*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D013171:Spores, Bacterial / Q000502:physiology; D013172:Spores, Fungal / Q000502:physiology; D013191:Stachybotrys / Q000472:pathogenicity*; D013302:Streptomyces / Q000472:pathogenicity*
D016207:Cytokines; D018836:Inflammation Mediators; D009569:Nitric Oxide; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases