title: Commercial peptidoglycan preparations are contaminated with superantigen-like activity that stimulates IL-17 production.
abstract: The immunomodulatory properties of peptidoglycan (PGN), a constituent of the bacterial cell wall, have been studied extensively but with contrasting results. Recent studies have demonstrated that the TLR2-mediated inflammatory responses elicited by Gram-positive PGN preparations are in fact a result of contaminating lipoproteins and lipoteichoic acid that can be removed only through sophisticated extraction procedures. Here, we report that commercial preparations of Staphylococcus aureus or Streptococcus pyogenes PGN are contaminated with bacterial superantigens (SAg). The T cell-derived cytokines IL-17A and IL-17F were induced by PGN preparations but not by TLR agonists or nucleotide-binding and oligomerization domain-like receptor agonists in human PBMC. IL-17 induction by PGN preparations was sensitive to protease digestion and required TCR signaling. Bacterial SAg could be detected by immunoblot in the PGN preparations, and purified recombinant SAg were powerful inducers of IL-17. Finally, the PGN preparations stimulated proliferation and expansion of T cells bearing specific TCR V beta elements. Our results suggest that a large body of literature that relied on commercial PGN preparations to study inflammatory diseases, such as arthritis, where IL-17 also plays an important role, should be interpreted with caution and possibly revisited. Future studies aimed at characterizing the activities of PGN should use PGN preparations of proven purity.
authors.affiliation: Department of Molecular Sciences, University of Tennessee Health Science Center, 858 Madison Avenue, Memphis, TN 38163, USA.
authors.forename: Hanfen
authors.initials: H
authors.lastname: Li
authors.forename: Mohammed M
authors.initials: MM
authors.lastname: Nooh
authors.forename: Malak
authors.initials: M
authors.lastname: Kotb
authors.forename: Fabio
authors.initials: F
authors.lastname: Re
mesh_terms: D000942:Antigens, Bacterial / Q000032:analysis* / Q000494:pharmacology; D016477:Artifacts; D002460:Cell Line; D004340:Drug Contamination; D005786:Gene Expression Regulation / Q000187:drug effects*; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D006801:Humans; D020381:Interleukin-17 / Q000096:biosynthesis* / Q000235:genetics; D008213:Lymphocyte Activation / Q000187:drug effects; D008222:Lymphokines / Q000096:biosynthesis / Q000235:genetics; D016328:NF-kappa B / Q000378:metabolism; D010457:Peptidoglycan / Q000032:analysis* / Q000302:isolation & purification; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis; D013211:Staphylococcus aureus / Q000737:chemistry*; D013297:Streptococcus pyogenes / Q000737:chemistry*; D018089:Superantigens / Q000032:analysis* / Q000494:pharmacology; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D051195:Toll-Like Receptor 2 / Q000819:agonists / Q000502:physiology
chemical_list: D000942:Antigens, Bacterial; C508717:IL17A protein, human; C440021:IL17F protein, human; D020381:Interleukin-17; D008222:Lymphokines; D016328:NF-kappa B; D010457:Peptidoglycan; D016693:Receptors, Antigen, T-Cell, alpha-beta; D018089:Superantigens; C495342:TLR2 protein, human; D051195:Toll-Like Receptor 2