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 (ECA PG and ECA LPS , respectively). Only the latter form is ECAimmunogenic. We previously demonstrated that Yersinia enterocolitica O : 3 and its rough (Ospecific polysaccharide-negative) mutants were ECA-immunogenic, suggesting that they contained ECA LPS ; 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 ECA PG and ECA LPS 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 ECA LPS -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 ECA LPS was linked to the Kdo region of LPS in Y. enterocolitica O : 3.

Generation of a CRISPR database for Yersinia pseudotuberculosis complex and role of CRISPR‐based immunity in conjugation

“…pTM100‐CRISPR was electroporated into E. coli strain ω7249. pTM100‐waaF (Noszczynska et al ., ) was used as spacer‐free control plasmid in mobilization experiments.…”

Interaction of human mannose-binding lectin (MBL) with Yersinia enterocolitica lipopolysaccharide

“…MBL was originally described as the murine Ra-reactive factor (RaRF), activating the complement cascade by interaction with Salmonella LPS of Ra type but not with the smooth or Re chemotypes (Ihara et al, 1982). Later, MBL has been demonstrated to react with OPS or YeO3-c-R1 waaF::cat-Mu, clmR Rd2 (LA-3/8IC) c Noszczynska et al (2015) a All the Y. enterocolitica strains are ECA-positive, except the ones from which the ECA-gene cluster has been deleted. b The OC hexasaccharide is truncated by two glucose residues.…”

“…The exact linkage of ECA LPS to the LA-IC (Fig. 1B) is not determined yet although it was indirectly shown to be the Kdo region (Noszczynska et al, 2015).…”

Lipopolysaccharide-Linked Enterobacterial Common Antigen (ECALPS) Occurs in Rough Strains of Escherichia coli R1, R2, and R4

“…E. coli O14 was proven later on to be rough and synthesize LOS characterized by the R4 core chemotype [ 13 , 14 ]. Further studies demonstrated that ECA LPS -dependent immunogenicity generally was limited only to a few rough enterobacteria with complete core OS, such as, for example, E. coli R1, R4, and K-12 [ 15 ], E. coli R2 and R3 [ 15 , 16 , 17 ], Proteus mirabilis [ 18 ], S. sonnei phase II [ 19 ], and Yersinia enterocolitica O:3 and O:9 [ 20 , 21 , 22 , 23 ].…”