Decisive role of lipopolysaccharide in activating nitric oxide and cytokine production by the probiotic Escherichia coli strain Nissle 1917.
Effects of Gram-negative probiotic E. coli strain Nissle 1917 (EcN) on the production of nitric oxide (NO) and cytokines were determined in cultures of resident peritoneal cells of rats. The cells (2 x 10(6)/mL) were cultured for 24 h in the presence of live EcN suspension (EcN-Susp), bacteria-free supernatant of this suspension (Sup-EcN), and LPS of EcN origin (LPS-EcN). The biosynthesis of NO was substantially enhanced using live bacteria counts as low as 10(3)/mL applied in the form of EcN-Susp. The same NO-enhancing effect was produced by the correspondingly diluted Sup-EcN. It was found that Sup-EcN contained relatively high amounts of LPS. Administration of the LPS-EcN mimicked the high NO-augmenting activities of both Sup-EcN and EcN-Susp. However, the activity of LPS-EcN was significantly less pronounced than were the activities of Sup-EcN and EcN-Susp containing identical amounts of LPS. The NO-stimulatory effects of the EcN preparations were completely inhibited by polymyxin B. All LPS-EcN and correspondingly diluted Sup-EcN and EcN-Susp stimulated the secretion of cytokines TNF-alpha, IL-1beta, IL-6, IL-10 and VEGF. Also these effects were abrogated by polymyxin B. In contrast to the effects on NO production, the cytokine-stimulatory effects were significantly less pronounced after the exposure of the cells to Sup-EcN and EcN-Susp than to the identical amounts of LPS-EcN. It may be concluded that the in vitro stimulatory effects of EcN on NO and cytokine production are mediated by LPS. It is suggested that the immunostimulatory activity of LPS is modulated by EcN-derived factor(s), the nature of which remains to be identified.
Zídek
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Institute of Experimental Medicine, v.v.i., Academy of Sciences of the Czech Republic, 142 20, Prague 4, Czech Republic. zidekz@biomed.cas.cz
Kmonícková
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Kostecká
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Tlaskalová-Hogenová
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D000818:Animals; D001424:Bacterial Infections / Q000276:immunology* / Q000382:microbiology; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology*; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000276:immunology* / Q000378:metabolism; D005260:Female; D006801:Humans; D008070:Lipopolysaccharides / Q000276:immunology* / Q000378:metabolism; D009569:Nitric Oxide / Q000276:immunology*; D010529:Peritoneal Cavity / Q000166:cytology / Q000382:microbiology; D019936:Probiotics; D051381:Rats; D017208:Rats, Wistar
D016207:Cytokines; D008070:Lipopolysaccharides; D009569:Nitric Oxide