title: Characterization of lipoteichoic acid structures from three probiotic Bacillus strains: involvement of D-alanine in their biological activity.
abstract: Probiotics represent a potential strategy to influence the host's immune system thereby modulating immune response. Lipoteichoic Acid (LTA) is a major immune-stimulating component of Gram-positive cell envelopes. This amphiphilic polymer, anchored in the cytoplasmic membrane by means of its glycolipid component, typically consists of a poly (glycerol-phosphate) chain with D-alanine and/or glycosyl substitutions. LTA is known to stimulate macrophages in vitro, leading to secretion of inflammatory mediators such as Nitric Oxide (NO). This study investigates the structure-activity relationship of purified LTA from three probiotic Bacillus strains (Bacillus cereus CH, Bacillus subtilis CU1 and Bacillus clausii O/C). LTAs were extracted from bacterial cultures and purified. Chemical modification by means of hydrolysis at pH 8.5 was performed to remove D-alanine. The molecular structure of native and modified LTAs was determined by (1)H NMR and GC-MS, and their inflammatory potential investigated by measuring NO production by RAW 264.7 macrophages. Structural analysis revealed several differences between the newly characterized LTAs, mainly relating to their D-alanylation rates and poly (glycerol-phosphate) chain length. We observed induction of NO production by LTAs from B. subtilis and B. clausii, whereas weaker NO production was observed with B. cereus. LTA dealanylation abrogated NO production independently of the glycolipid component, suggesting that immunomodulatory potential depends on D-alanine substitutions. D-alanine may control the spatial configuration of LTAs and their recognition by cell receptors. Knowledge of molecular mechanisms behind the immunomodulatory abilities of probiotics is essential to optimize their use.
authors.affiliation: Laboratoire de Chimie des Substances Naturelles, EA 1069, Antenne IUT, Département Génie Biologique, Allée André Maurois, 87065, Limoges, France.
authors.forename: Romain
authors.initials: R
authors.lastname: Villéger
authors.forename: Naima
authors.initials: N
authors.lastname: Saad
authors.forename: Karine
authors.initials: K
authors.lastname: Grenier
authors.forename: Xavier
authors.initials: X
authors.lastname: Falourd
authors.forename: Loïc
authors.initials: L
authors.lastname: Foucat
authors.forename: Maria C
authors.initials: MC
authors.lastname: Urdaci
authors.forename: Philippe
authors.initials: P
authors.lastname: Bressollier
authors.forename: Tan-Sothea
authors.initials: TS
authors.lastname: Ouk
mesh_terms: D000409:Alanine / Q000032:analysis* / Q000276:immunology*; D000818:Animals; D001407:Bacillus / Q000737:chemistry* / Q000276:immunology; D002460:Cell Line; D008401:Gas Chromatography-Mass Spectrometry; D006868:Hydrolysis; D007155:Immunologic Factors / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D008070:Lipopolysaccharides / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*; D008264:Macrophages / Q000187:drug effects; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D015394:Molecular Structure; D009569:Nitric Oxide / Q000378:metabolism; D019936:Probiotics / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013682:Teichoic Acids / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*
chemical_list: D007155:Immunologic Factors; D008070:Lipopolysaccharides; D013682:Teichoic Acids; D009569:Nitric Oxide; C009900:lipoteichoic acid; D000409:Alanine
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