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

Kishimoto M, Nomoto R, Osawa R - Biosci Microbiota Food Health (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioresource Science, Organization of Advanced Science and Technology, Kobe University, 1-1 Rokko-dai, Nada-ku, Kobe 657-8501, Japan.

ABSTRACT
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.

No MeSH data available.


Thin layer chromatography analysis of sugar compositions of each EPS.Lane 1, glucose (0.5%, wt/vol); lane 2, galactose (0.5%, wt/vol); lane 3, glucose+galactose (1:1); lane 4, EPS of L. delbrueckii TU-1; lane 5, EPS of L. delbrueckii JCM1002T; lane 6, EPS of L. delbrueckii JCM1012T; lane 7, EPS of L. delbrueckii JCM1248T; lane 8, EPS of L. delbrueckii JCM15610T; lane 9, EPS of L. delbrueckii KM-1; lane 10, EPS of L. delbrueckii KM-2; lane 11, EPS of L. delbrueckii KM-3; lane 12, EPS of L. delbrueckii KM-4; lane 13, EPS of L. delbrueckii KM-5. Development system: isopropanol/ethyl acetate/water (3:1:1) solvent.
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fig_001: Thin layer chromatography analysis of sugar compositions of each EPS.Lane 1, glucose (0.5%, wt/vol); lane 2, galactose (0.5%, wt/vol); lane 3, glucose+galactose (1:1); lane 4, EPS of L. delbrueckii TU-1; lane 5, EPS of L. delbrueckii JCM1002T; lane 6, EPS of L. delbrueckii JCM1012T; lane 7, EPS of L. delbrueckii JCM1248T; lane 8, EPS of L. delbrueckii JCM15610T; lane 9, EPS of L. delbrueckii KM-1; lane 10, EPS of L. delbrueckii KM-2; lane 11, EPS of L. delbrueckii KM-3; lane 12, EPS of L. delbrueckii KM-4; lane 13, EPS of L. delbrueckii KM-5. Development system: isopropanol/ethyl acetate/water (3:1:1) solvent.

Mentions: The monosaccharide composition of each EPS produced by L. delbrueckii strains was evaluated by TLC analysis. A dark oval-shaped glucose spot was seen in the upper part of the plate (Fig. 1Fig. 1.


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

Kishimoto M, Nomoto R, Osawa R - Biosci Microbiota Food Health (2014)

Thin layer chromatography analysis of sugar compositions of each EPS.Lane 1, glucose (0.5%, wt/vol); lane 2, galactose (0.5%, wt/vol); lane 3, glucose+galactose (1:1); lane 4, EPS of L. delbrueckii TU-1; lane 5, EPS of L. delbrueckii JCM1002T; lane 6, EPS of L. delbrueckii JCM1012T; lane 7, EPS of L. delbrueckii JCM1248T; lane 8, EPS of L. delbrueckii JCM15610T; lane 9, EPS of L. delbrueckii KM-1; lane 10, EPS of L. delbrueckii KM-2; lane 11, EPS of L. delbrueckii KM-3; lane 12, EPS of L. delbrueckii KM-4; lane 13, EPS of L. delbrueckii KM-5. Development system: isopropanol/ethyl acetate/water (3:1:1) solvent.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300312&req=5

fig_001: Thin layer chromatography analysis of sugar compositions of each EPS.Lane 1, glucose (0.5%, wt/vol); lane 2, galactose (0.5%, wt/vol); lane 3, glucose+galactose (1:1); lane 4, EPS of L. delbrueckii TU-1; lane 5, EPS of L. delbrueckii JCM1002T; lane 6, EPS of L. delbrueckii JCM1012T; lane 7, EPS of L. delbrueckii JCM1248T; lane 8, EPS of L. delbrueckii JCM15610T; lane 9, EPS of L. delbrueckii KM-1; lane 10, EPS of L. delbrueckii KM-2; lane 11, EPS of L. delbrueckii KM-3; lane 12, EPS of L. delbrueckii KM-4; lane 13, EPS of L. delbrueckii KM-5. Development system: isopropanol/ethyl acetate/water (3:1:1) solvent.
Mentions: The monosaccharide composition of each EPS produced by L. delbrueckii strains was evaluated by TLC analysis. A dark oval-shaped glucose spot was seen in the upper part of the plate (Fig. 1Fig. 1.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioresource Science, Organization of Advanced Science and Technology, Kobe University, 1-1 Rokko-dai, Nada-ku, Kobe 657-8501, Japan.

ABSTRACT
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.

No MeSH data available.