title: Serological characterization of the enterobacterial common antigen substitution of the lipopolysaccharide of Yersinia enterocolitica O : 3.
abstract: Enterobacterial common antigen (ECA) is a polysaccharide present in all members of Enterobacteriaceae anchored either via phosphatidylglycerol (PG) or LPS to the outer leaflet of the outer membrane (ECAPG and ECALPS, respectively). Only the latter form is ECA-immunogenic. We previously demonstrated that Yersinia enterocolitica O : 3 and its rough (O-specific polysaccharide-negative) mutants were ECA-immunogenic, suggesting that they contained ECALPS; however, it was not known which part of the LPS core region was involved in ECA binding. To address this, we used a set of three deep-rough LPS mutants for rabbit immunization. The polyvalent antisera obtained were: (i) analysed for the presence of anti-LPS and anti-ECA antibodies; (ii) treated with caprylic acid (CA) to precipitate IgM antibodies and protein aggregates; and (iii) adsorbed with live ECA-negative bacteria to obtain specific anti-ECA antisera. We demonstrated the presence of antibodies specific for both ECAPG and ECALPS in all antisera obtained. Both CA treatment and adsorption with ECA-negative bacteria efficiently removed anti-LPS antibodies, resulting in specific anti-ECA sera. The LPS of the ECALPS-positive deepest-rough mutant contained only lipid A and 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) residues of the inner core, suggesting that ECALPS was linked to the Kdo region of LPS in Y. enterocolitica O : 3.
authors.affiliation: Department of Microbiology, University of Silesia, Jagiellońska 28, PL- 40-032 Katowice, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Noszczyńska
authors.affiliation: Department of Microbiology, University of Silesia, Jagiellońska 28, PL- 40-032 Katowice, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kasperkiewicz
authors.affiliation: Division of Structural Biochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany.
authors.forename: Katarzyna Anna
authors.initials: KA
authors.lastname: Duda
authors.affiliation: Division of Structural Biochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany.|Department of Microbiology, University of Silesia, Jagiellońska 28, PL- 40-032 Katowice, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Podhorodecka
authors.affiliation: Department of Microbiology, University of Silesia, Jagiellońska 28, PL- 40-032 Katowice, Poland.
authors.forename: Kamila
authors.initials: K
authors.lastname: Rabsztyn
authors.affiliation: Department of Microbiology, University of Silesia, Jagiellońska 28, PL- 40-032 Katowice, Poland.
authors.forename: Karolina
authors.initials: K
authors.lastname: Gwizdała
authors.affiliation: Department of Immunobiology of Infections, Institute of Medical Biology, PAS, Lodowa 106, PL- 93-232 Łódź, Poland.
authors.forename: Anna Stanisława
authors.initials: AS
authors.lastname: Świerzko
authors.affiliation: Department of Microbiology, University of Silesia, Jagiellońska 28, PL- 40-032 Katowice, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Radziejewska-Lebrecht
authors.affiliation: Division of Structural Biochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany.
authors.forename: Otto
authors.initials: O
authors.lastname: Holst
authors.affiliation: Helsinki University Central Hospital Laboratory Diagnostics, FIN-00270 Helsinki, Finland.|Department of Bacteriology and Immunology, Haartman Institute, and Research Programs Unit, Immunobiology, University of Helsinki, FIN-00014, Helsinki, Finland.
authors.forename: Mikael
authors.initials: M
authors.lastname: Skurnik
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology*; D007106:Immune Sera; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology*; D009154:Mutation; D011817:Rabbits; D012703:Serotyping; D015008:Yersinia enterocolitica / Q000145:classification / Q000235:genetics / Q000276:immunology*
chemical_list: D000906:Antibodies; D000942:Antigens, Bacterial; D007106:Immune Sera; D008070:Lipopolysaccharides