Effects of Live and Heat-Inactivated E. coli Strains and Their Supernatants on Immune Regulation in HT-29 Cells.

Christian Zimmermann, Marius Schild, Clemens Kunz, Kurt Zimmermann, Sabine Kuntz
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引用次数: 8

Abstract

Probiotics are considered to have a beneficial impact on humans, but in some cases, administration of live microorganisms might be risky. In the present study, immunomodulatory effects of different Escherichia coli strains and their super-natants were examined under different inflammatory conditions with living and heat-inactivated strains. HT-29 cells were incubated with E. coli strains (S2-G1, S2-G3, S2-G4 and S2-G8) and their supernatants with or without stimulation with tumor necrosis factor alpha (TNF-α) or interleukin (IL)-1β. Quantification of IL-8 secretion and gene expression was performed by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR). IL-8 secretion by TNF-α- and IL-1β-stimulated cells was attenuated by all four live strains. In contrast, heat inactivation resulted in an elevated IL-8 expression and secretion in unstimulated cells and did not maintain the anti-inflammatory effect of live bacteria in cytokine-stimulated cells. The supernatant of the live S2-G3 led to an elevated IL-8 secretion in unstimulated and IL-1β-stimulated cells but not in TNF-α-stimulated cells. Live bacteria of all strains might induce an immunosuppressive effect after stimulation of HT-29 cells, whereas heat inactivation and the supernatant seem to induce an elevated immune response. These findings might have an impact depending on the indication and purpose of administration.

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活的和热灭活的大肠杆菌菌株及其上清液对HT-29细胞免疫调节的影响。
益生菌被认为对人类有有益的影响,但在某些情况下,使用活微生物可能有风险。在本研究中,用活菌株和热灭活菌株在不同炎症条件下检测了不同大肠杆菌菌株及其上清液的免疫调节作用。HT-29细胞与大肠杆菌菌株(S2-G1、S2-G3、S2-G4和S2-G8)及其上清液在肿瘤坏死因子-α(TNF-α)或白细胞介素-1β刺激或不刺激下孵育。通过酶联免疫吸附试验(ELISA)和实时聚合酶链式反应(PCR)对IL-8分泌和基因表达进行定量。TNF-α和IL-1β刺激的细胞分泌的IL-8被所有四种活菌株减弱。相反,热失活导致未刺激细胞中IL-8的表达和分泌升高,并且在细胞因子刺激的细胞中没有维持活细菌的抗炎作用。活S2-G3的上清液导致未刺激和IL-1β刺激的细胞中IL-8分泌增加,但TNF-α刺激的细胞不分泌。所有菌株的活细菌在刺激HT-29细胞后可能诱导免疫抑制作用,而热灭活和上清液似乎诱导免疫反应升高。这些发现可能会产生影响,这取决于给药的适应症和目的。
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