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In vitro evaluation of immunological properties of extracellular polysaccharides produced by Lactobacillus delbrueckii strains

机译:德氏乳杆菌菌株产生的细胞外多糖的免疫学特性的体外评估

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摘要

We investigated the variation in immunological properties of the extracellular polysaccharides (EPSs) produced by different Lactobacillus delbrueckii strains as well as that of their monosaccharide composition. The monosaccharide composition of each EPS produced by L. delbrueckii strains, as determined by thin layer chromatography (TLC), showed an appreciable variation in a strain-dependent manner, which could be broadly assigned to 4 TLC groups. Meanwhile, the immunological properties of the EPSs produced by 10 L. delbrueckii strains were evaluated in a semi-intestinal model using a Transwell co-culture system, which employed human intestinal epithelial Caco-2 cells on the apical side and murine macrophage RAW264.7 cells on the basolateral side. Each EPS was added to the apical side to allow direct contact with Caco-2 cells and incubated for 6 hr. After incubation, the amounts of TNF-α and several cytokines that had been released by either RAW264.7 or Caco-2 cells were then quantified by cytotoxic activity on L929 cells or the RT-PCR method. It was found that the EPS-stimulated RAW264.7 cells express different profiles of cytokine production via Caco-2 cells but that the profile difference could not be related to the above TLC grouping. The evidence suggests that the EPSs of L. delbrueckii strains are diverse not only in their biochemical structure but also in their immunological properties.
机译:我们调查了由不同的德氏乳杆菌菌株产生的细胞外多糖(EPS)及其单糖组成的免疫学特性的变化。通过薄层色谱法(TLC)测定,由德氏乳杆菌菌株产生的每种EPS的单糖组成均以菌株依赖性方式显示出明显的变化,该变化可大致分为4个TLC组。同时,使用Transwell共培养系统在半肠道模型中评估了10个德氏乳杆菌菌株产生的EPS的免疫学特性,该系统在顶侧使用人肠道上皮Caco-2细胞并使用鼠巨噬细胞RAW264.7细胞在基底外侧。将每种EPS添加至顶端,以使其与Caco-2细胞直接接触,并孵育6小时。孵育后,然后通过对L929细胞的细胞毒活性或RT-PCR方法对RAW264.7或Caco-2细胞释放的TNF-α和几种细胞因子的量进行定量。发现EPS刺激的RAW264.7细胞表达通过Caco-2细胞产生的细胞因子的不同谱,但是该谱差异与上述TLC分组无关。有证据表明,德氏乳杆菌菌株的EPS不仅在生化结构上而且在免疫学特性上也各不相同。

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