Structural and Serological Characterization of the O Antigen of Proteus mirabilis Clinical Isolates Classified into a New Proteus Serogroup, O84.

Two closely related Proteus mirabilis smooth strains, Kr1 and Ks20, were isolated from wound and skin samples, respectively, of two infected patients in central Poland. Serological tests, using the rabbit Kr1-specific antiserum, revealed that both strains presented the same O serotype. Their O antigens are unique among the Proteus O serotypes, which had been described earlier, as they were not recognized in an enzyme-linked immunosorbent assay (ELISA) by a set of Proteus O1-O83 antisera. Additionally, the Kr1 antiserum did not react with O1-O83 lipopolysaccharides (LPSs). The O-specific polysaccharide (OPS, O antigen) of P. mirabilis Kr1 was obtained via the mild acid degradation of the LPSs, and its structure was established via a chemical analysis and one- and two-dimensional 1H and 13C nuclear magnetic resonance (NMR) spectroscopy applied to both initial and O-deacetylated polysaccharides, where most ß-2-acetamido-2-deoxyglucose (N-acetylglucosamine) (GlcNAc) residues are non-stoichiometrically O-acetylated at positions 3, 4, and 6 or 3 and 6, and a minority of α-GlcNAc residues are 6-O-acetylated. Based on the serological features and chemical data, P. mirabilis Kr1 and Ks20 were proposed as candidates to a new successive O-serogroup in the genus Proteus, O84, which is another example of new Proteus O serotypes identified lately among serologically differentiated Proteus bacilli infecting patients in central Poland.

The unique structure of bacterial polysaccharides - Immunochemical studies on the O-antigen of Proteus penneri 4034-85 clinical strain classified into a new O83 Proteus serogroup.

The serological classification scheme of the opportunistic Proteus bacilli includes a number of Proteus penneri strains. The tested P. penneri 4034-85 strain turned out to be serologically distinguished in ELISA and Western blotting. The O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of this strain and studied by sugar and methylation analyses and dephosphorylation along with 1H and 13C NMR spectroscopy, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C HSQC, HMBC, and HSQC-TOCSY experiments, The O-polysaccharide was found to have a linear repeating unit containing glycerol 1-phosphate and two residues each of Gal and GlcNAc. The following O-polysaccharide structure was established, which, to our knowledge, is unique among known bacterial polysaccharide structures.

Structural and serological characterization of the O82 antigen of a Proteus mirabilis strain isolated from a patient in Poland.

P. mirabilis strains Kro 45 and Kwy 46 were isolated from the pus and the muscular fluid, respectively, of a hospitalized 61-year-old female in Lódz, Poland. Both strains demonstrated a good swarming ability on a solid medium, and the Dienes test for differentiation of swarming strains indicated their identity. The strains were serologically identical and did not belong to any of the known Proteus O1-O81 serogroups. In this work, we studied the O-specific polysaccharide (O antigen) of P. mirabilis Kwy46, which defines the immunospecificity of the strain. The O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide, and the following structure of its oligosaccharide repeat (O-unit) was established by sugar analysis along with 1D and 2D 1H and 13C NMR spectroscopy: where (S)-lac indicates an (S)-1-carboxyethyl group [an (S)-lactic acid residue], which forms an ether with a GlcNAc residue (so called glycolactilic acid). This structure is unique among Proteus O-polysaccharides but shares a trisaccharide fragment with that of P. mirabilis O5. Studies of the cross-reactivity between P. mirabilis Kwy 46 O antiserum/lipopolysaccharide and Proteus O1-O81 lipopolysaccharides/O antisera allowed identification of a putative Kwy 46 O-antigen epitope. Based on the data obtained, it is proposed to create a new O82 serogroup within the genus Proteus represented by the studied P. mirabilis isolates.

Classification of a Proteus penneri clinical isolate with a unique O-antigen structure to a new Proteus serogroup, O80.

Proteus penneri is an opportunistic pathogen, which may cause severe diseases, most frequently urinary tract infections in immunocompromised patients. P. penneri Br 114 exhibiting a good swarming growth ability as an S-form strain was isolated from a wound of a patient in Lódz, Poland. Serological studies using ELISA and Western blotting and chemical analyses along with (1)H and (13)C NMR spectroscopy showed that the O-antigen (O-polysaccharide) of this strain is unique among the known Proteus serotypes O1-O79. It possesses a linear pentasaccharide repeating unit containing a partially O-acetylated amide of D-glucuronic acid (GlcA) with L-serine having the following structure: [structure: see text]. These data are a basis for creating a new Proteus serogroup, O80, so far represented by the single Br 114 isolate. The O80 is the 21st O-serogroup containing P. penneri strains and the fourth serogroup based on Proteus spp. clinical isolates from Lódz, Poland.

Structure and gene cluster of the O-antigen of Escherichia coli O36.

The O-polysaccharide (O-antigen) of Escherichia coli O36 was isolated from the lipopolysaccharide and studied by sugar analyses and Smith degradation along with (1)H and (13)C NMR spectroscopy. The following structure of the branched pentasaccharide repeating unit was established, which is unique among the known structures of bacterial polysaccharides: The O-antigen gene cluster of E. coli O36 has been sequenced. The gene functions were tentatively assigned by comparison with sequences in the available databases and found to be in full agreement with the O-polysaccharide structure.