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1

Serological classification and epitope specificity of Proteus penneri S29 lipopolysaccharide

100%

Zych K. , Siwinska M. , Sidorczyk Z.

Archivum Immunologiae et Therapiae Experimentalis

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2005

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tom 53

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nr 6

540-545

EN

Gram-negative bacteria of genus Proteus are common human intestinal and urinary tract pathogens. In the genus Proteus there are four clinically important named species: P. mirabilis, P. vulgaris, P. penneri, and P. hauseri, and three unnamed Proteus genomospecies: 4, 5, and 6. The clinical significance of P. penneri, described in 1982 as a new species, is poorly documented. The aim of this work is serological characterization and classification of a ceftriaxone-susceptible P. penneri S29 strain isolated from a 34-year-old patient with postneurosurgical meningitis. In this characterization we will also include a ceftriaxon-resistant strain, P. penneri R15, isolated from the same patient after 12 days' treatment with ceftriaxon and other antibiotics. Rabbit polyclonal O-antisera were obtained against these two strains and purified lipopolysaccharides (LPS) were extracted from the bacterial mass of the P. penneri S29 and R15 strains. In the serological investigations the following tests were used: enzyme immunosorbent assay (EIA), passive immunohemolysis (PIH), inhibition of these tests, absorption of rabbit O-antisera with the respective LPS, and repeated PIH, SDS/PAGE, and Western blot techniques. The serological studies of the LPS extracted from both P. penneri strains showed the identity of both preparations of O-polysaccharides from LPS.. In P. penneri S29 O-antiserum, four different types of antibodies were described and characterized. Both investigated P. penneri S29 and R15 strains were classified to the Proteus O31ab serogroup.

2

Serological classification and epitope specificity of Proteus vulgaris TG 251 from Proteus serogroup O65

88%

Zych K. , Kolodziejska K. , Drzewiecka D. , Perepelov A. V. , Knirel Y. A. , Sidorczyk Z.

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nr 3

187-191

EN

Introduction: Proteus rods are currently subdivided into five named species, i.e. Proteus mirabilis, P. vulgaris, P. penneri, P. hauseri, and P. myxofaciens, and three unnamed Proteus genomospecies 4 to 6. Based on the serospecificity of the lipopolysaccharide (LPS; O-antigen), strains of P. mirabilis and P. vulgaris were divided into 49 O-serogroups and 11 additional O-serogroups were proposed later. About 15 further O-serogroups have been proposed for the third medically important species, P. penneri. Here the serological classification of P. vulgaris strain TG 251, which does not belong to these serogroups, is reported. Serological investigations also allowed characterization of the epitope specificity of its LPS. Materials and Methods: Purified LPSs from five Proteus strains were used as antigens in enzyme immunosorbent assay (EIA), SDS/PAGE, and Western blot and alkali-treated LPSs in the passive immunohemolysis (PIH) test, inhibition of PIH and EIA, and absorption of the rabbit polyclonal O-antisera with the respective LPS. Results: The serological studies of P. vulgaris TG 251 LPS indicated the identity of its O-polysaccharide with that of P. penneri O65. The antibody specificities of P. vulgaris TG 251 and P. penneri O65 O-antisera, were described. Conclusions: P. vulgaris TG 251 was classified to the Proteus O65 serogroup. Two disaccharide-associated epitopes present in P. vulgaris TG 251 and P. penneri O65 LPSs are suggested to be responsible for cross-reactions with three heterologous Proteus strains.

3

Structure of the O-polysaccharide of Proteus mirabilis O19 and reclassification of certain Proteus strains that were formerly classified in serogroup O19

88%

Perepelov A. V. , Rozalski A. , Bartodziejska B. , Senchenkova S. N. , Knirel Y. A.

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2004

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tom 52

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nr 3

188-196

EN

Bacteria of the genus Proteus are a common cause of urinary tract infections. The O-polysaccharide chain of their LPS (O-antigen) defines the serological specificity of these bacteria. Based on the immunospecificity of the O-antigens, two species, P. mirabilis and P. vulgaris, were classified into 49 O-serogroups, and more O-serogroups for strains of these species and P. penneri have been subsequently proposed. The lipopolysaccharide of P. mirabilis CCUG 19011 from serogroup O19 was degraded and under mildly acidic and mildly alkaline conditions. Polysaccharides thus obtained were studied by chemical methods, including O-deacetylation, sugar and methylation analyses, and 1H- and 13C-NMR spectroscopy. Antisera were obtained by immunization of New Zealand white rabbits with heat-killed bacteria. In serological studies, enzyme immunosorbent assay, passive hemolysis test, and inhibition of passive hemolysis were used.The following structure of the O-polysaccharide repeating unit was established. ->3)-beta-D-GlcpNAc-(1->3)alpha-D-GalpNAc4,6(R-Pyr)-(1->4)-alpha-D-GalpA-(1->3)-alpha-L-Rhap2Ac-(1-> where R-Pyr is (R)-1-carboxyethylidene (an acetal-linked pyruvic acid). This structure is significantly different from the O-polysaccharide structures of P. vulgaris, P. hauseri and P. penneri strains from the same Proteus serogroup O19. Conclusions: Based on immunochemical studies of the lipopolysaccharides, it is suggested 1) to keep P. vulgaris CCUG 4654 and P. penneri 31 in serogroup O19 as two subgroups, 2) to reclassify P. mirabilis CCUG 19011 into a new Proteus serogroup, O51, and 3) to classify serologically related strains, including P. vulgaris ATCC 49990, P. hauseri 1732-80 and 1086-80, P. penneri 15, and some other P. penneri strains, in yet another Proteus serogroup, O52. This structure is significantly different from the O-polysaccharide structures of P. vulgaris, P. hauseri and P. penneri strains from the same Proteus serogroup O19. Based on immunochemical studies of the lipopolysaccharides, it is suggested 1) to keep P. vulgaris CCUG 4654 and P. penneri 31 in serogroup O19 as two subgroups, 2) to reclassify P. mirabilis CCUG 19011 into a new Proteus serogroup, O51, and 3) to classify serologically related strains, including P. vulgaris ATCC 49990, P. hauseri 1732-80 and 1086-80, P. penneri 15, and some other P. penneri strains, in yet another Proteus serogroup, O52.

4

Characterization and serological classification of a collection of Proteus penneri clinical strains

88%

Drzewiecka D. , Zych K. , Sidorczyk Z.

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nr 2

121-128

EN

Chronic Granulomatous disease bacteria of the genus Proteus, which are a common cause of urinary tract infections, are divided into four species: P. mirabilis, P. vulgaris, P. penneri, and P. hauseri, and three unnamed genomospecies, Proteus 4, 5, and 6 (single-strain species P. myxofaciens was isolated from the gypsy moth). Establishing the serological classification of these species would aid in completing the classification scheme of the whole genus Proteus and in applying serological methods in diagnostic procedures and epidemiological investigations for these opportunistic pathogens. The aim of this research was a serological characterization and classification of 57 Proteus penneri clinical strains, isolated from patients from different countries all over the world, into Proteus O serogroups. Purified lipopolysaccharides (LPSs) extracted from 57 P. penneri strains were used as antiandgens in enzyme immunosorbent assay (EIA), SDS/PAGE, and Western blot techniques, and alkali-treated LPSs in passive immunohemolysis test (PIH), inhibition of PIH, and absorption of rabbit polyclonal O-antisera. Results: That result confirms the serological distinction of this species within the genus Proteus, and may have diagnostic significance. Conclusions: As a result of serological studies of LPSs extracted from the P. penneri strains, one new Proteus serogroup, represented by the P. penneri 97 strain, was established. Three further strains were classified into the Proteus serogroup O8, which had not contained any P. penneri strains before. All the remaining strains were classified into 11 already existing Proteus O serogroups. It is important to emphasize that 72% of studied strains were classified into serogroups that contain P. penneri strains only.