title: Lipopeptide structure determines TLR2 dependent cell activation level.
abstract: Bacterial lipoproteins/peptides are composed of di-O-acylated-S-(2,3-dihydroxypropyl)-cysteinyl residues N-terminally coupled to distinct polypeptides, which can be N-acylated with a third fatty acid. Using a synthetic lipopeptide library we characterized the contribution of the lipid portion to the TLR2 dependent pattern recognition. We found that the two ester bound fatty acid length threshold is beyond eight C atoms because almost no response was elicited by cellular challenge with analogues carrying shorter acyl chains in HEK293 cells expressing recombinant human TLR2. In contrast, the amide bound fatty acid is of lesser importance. While two ester-bound palmitic acids mediate a high stimulatory activity of the respective analogue, a lipopeptide carrying one amide-bound and another ester-bound palmitic acid molecule was inactive. In addition, species specific LP recognition through murine and human TLR2 depended on the length of the two ester bound fatty acid chains. In conclusion, our results indicate the responsibility of both ester bound acyl chains but not of the amide bound fatty acid molecule for the TLR dependent cellular recognition of canonical triacylated LP, as well as a requirement for a minimal acyl chain length. Thus they might support the explanation of specific immuno-stimulatory potentials of different microorganisms and provide a basis for rational design of TLR2 specific adjuvants mediating immune activation to distinct levels.
authors.affiliation: Department of Immunology and Cell Biology, Research Center Borstel, Borstel, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Buwitt-Beckmann
authors.forename: Holger
authors.initials: H
authors.lastname: Heine
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Wiesmüller
authors.forename: Günther
authors.initials: G
authors.lastname: Jung
authors.forename: Roland
authors.initials: R
authors.lastname: Brock
authors.forename: Artur J
authors.initials: AJ
authors.lastname: Ulmer
mesh_terms: D000215:Acylation; D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D001426:Bacterial Proteins; D002460:Cell Line; D005227:Fatty Acids / Q000737:chemistry; D006801:Humans; D007109:Immunity / Q000187:drug effects; D016209:Interleukin-8 / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry* / Q000494:pharmacology*; D051379:Mice; D013329:Structure-Activity Relationship; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000502:physiology*; D014162:Transfection
chemical_list: D000276:Adjuvants, Immunologic; D001426:Bacterial Proteins; D005227:Fatty Acids; D016209:Interleukin-8; D008074:Lipoproteins; C495342:TLR2 protein, human; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2