Serological characterization of the enterobacterial common antigen substitution of the lipopolysaccharide of 'Yersinia enterocolitica' O:3
| Autor: | Joanna Radziejewska-Lebrecht, Karolina Gwizdała, Anna Stanisława Świerzko, Mikael Skurnik, Katarzyna A. Duda, Joanna Podhorodecka, Kamila Rabsztyn, Otto Holst, Magdalena Noszczyńska, Katarzyna Kasperkiewicz |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Lipopolysaccharides
Lipopolysaccharide electrospray ionization whole-cell lysate Microbiology Antibodies Lipid A enterobacterial common antigen chemistry.chemical_compound Antigen Animals Serotyping Yersinia enterocolitica Antiserum Antigens Bacterial biology Immune Sera caprylic acid biology.organism_classification Enterobacteriaceae 3. Good health Yersinia enterocolitica O : 3 chemistry Mutation biology.protein lipids (amino acids peptides and proteins) Rabbits Antibody Bacterial outer membrane |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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