title: Platelet factor 4 binding to lipid A of Gram-negative bacteria exposes PF4/heparin-like epitopes.
abstract: The positively charged chemokine platelet factor 4 (PF4) forms immunogenic complexes with heparin and other polyanions. Resulting antibodies can induce the adverse drug effect heparin-induced thrombocytopenia. PF4 also binds to bacteria, thereby exposing the same neoantigen(s) as with heparin. In this study, we identified the negatively charged lipopolysaccharide (LPS) as the PF4 binding structure on Gram-negative bacteria. We demonstrate by flow cytometry that mutant bacteria with progressively truncated LPS structures show increasingly enhanced PF4 binding activity. PF4 bound strongest to mutants lacking the O-antigen and core structure of LPS, but still exposing lipid A on their surfaces. Strikingly, PF4 bound more efficiently to bisphosphorylated lipid A than to monophosphorylated lipid A, suggesting that phosphate residues of lipid A mediate PF4 binding. Interactions of PF4 with Gram-negative bacteria, where only the lipid A part of LPS is exposed, induce epitopes on PF4 resembling those on PF4/heparin complexes as shown by binding of human anti-PF4/heparin antibodies. As both the lipid A on the surface of Gram-negative bacteria and the amino acids of PF4 contributing to polyanion binding are highly conserved, our results further support the hypothesis that neoepitope formation on PF4 after binding to bacteria is an ancient host defense mechanism.
authors.affiliation: Institut für Immunologie und Transfusionsmedizin, Greifswald, Germany.
authors.forename: Krystin
authors.initials: K
authors.lastname: Krauel
authors.forename: Claudia
authors.initials: C
authors.lastname: Weber
authors.forename: Sven
authors.initials: S
authors.lastname: Brandt
authors.forename: Ulrich
authors.initials: U
authors.lastname: Zähringer
authors.forename: Uwe
authors.initials: U
authors.lastname: Mamat
authors.forename: Andreas
authors.initials: A
authors.lastname: Greinacher
authors.forename: Sven
authors.initials: S
authors.lastname: Hammerschmidt
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D000939:Epitopes / Q000276:immunology* / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006493:Heparin / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D008050:Lipid A / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000235:genetics / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D019081:O Antigens / Q000276:immunology; D010587:Phagocytosis; D010978:Platelet Factor 4 / Q000276:immunology* / Q000378:metabolism; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013921:Thrombocytopenia / Q000276:immunology* / Q000378:metabolism
chemical_list: D000939:Epitopes; D008050:Lipid A; D008070:Lipopolysaccharides; D019081:O Antigens; D010978:Platelet Factor 4; D006493:Heparin