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1

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

100%
Perepelov A. V., Rozalski A., Bartodziejska B., Senchenkova S. N., Knirel Y. A.
Archivum Immunologiae et Therapiae Experimentalis
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2004
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vol. 52
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issue 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.
2

Serological characterization of the O-specific polysaccharide of Providencia alcalifaciens O23

100%
Torzewska A., Kocharova A. N., Maszewska A., Knirel Y. A., Rozalski A.
Archivum Immunologiae et Therapiae Experimentalis
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2004
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vol. 52
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issue 1
43-49
EN
The genus Providencia belongs to the Enterobacteriaceae family and currently consists of five species: P.alcalifaciens, P.heimbachae, P.rettgerii, P.rustigianii and P.stuartii .The serological classification scheme of P.alcalifaciens, P.rustigianii and P.stuartii includes 63 O-serogroups and 30 H-serogroups. The O-antigenic specificity is defined by the structure of the O-antigen(O-specific polysaccharide ?OPS), a part of the lipopolysaccharide (LPS,endotoxin),one of the major components of the outer membrane of Gram-negative bacteria and an important virulence factor of these bacteria. Among the bacteria of the Enterobacteriaceae family, the genus Providencia is one of the least studied in respect to its LPS structure and antigenic specificity. Studies of the chemical structures and the serological specificity of the O-antigens aim at the elucidation of the molecular basis of the serological classification of Providenciasp. LPS and alkali-treated LPS of P.alcalifaciens O23 and serologically related P.rustigianii O14, P.mirabilis O13 and P.myxofaciens as well as O-antiserum against P.alcalifaciens O23 were used. Serological characterization of P.alcalifaciens O23 O-specific polysaccharide was done by use enzyme immunosorbent assay (EIA), passive hemolysis test (PHT)as well as by inhibition and sodium deoxycholate polyacrylamide gel electrophoresis (DOC-PAGE)of LPS and Western blot. The OPS of P.alcalifaciens,O23,contains an N-(D-glucuronoyl)-N-[(R)-1-carboxyethyl ]-L- lysine residue (GlcAAlaLys).The LPS of P.alcalifaciens,O23,and other LPSs containing AlaLys from Providencia and Proteus strains were tested with rabbit anti-P.alcalifiaciens O23 serum. The serological data showed that a GlcAAlaLys-associated epitope plays a role as an antigenic determinant in the P.alcalifaciens O23 OPS and revealed the particular importance of glucuronic acid and the carboxyethyl group for the binding of O23-specific antibodies.
3

Structures and serology of the O-specific polysaccharides of bacteria of the genus Citrobacter

100%
Knirel Y. A., Kocharova N. A., Bystrova O. V., Katzenellenbogen E., Gamian A.
Archivum Immunologiae et Therapiae Experimentalis
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2002
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vol. 50
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issue 6
377-389
EN
The review presents the structures of the O-specific polysaccharides (O-antigens) of the lipopolysaccharides isolated from over 25 Citrobacter strains, which represent different species and serogroups. The correlation between O-antigen structure and immunospecificity as well as numerous cross-reactions between Citrobacter and other enterobacterial species are discussed.
4

Immunochemical studies on the O-antigens of Proteus mirabilis O23 and Proteus vulgaris O23

86%
Rozalski A., Perepelov A. V., Bartodziejska B., Senchenkova S. N., Babicka D., Kaca W., Knirel Y. A.
Archivum Immunologiae et Therapiae Experimentalis
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2003
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vol. 51
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issue 1
69-73
EN
Analysis by 1H and 13C nuclear magnetic resonance (NMR) spectroscopy demonstrated that the O-specific polysaccharides of P. mirabilis PrK 42/57 and P. vulgaris PrK 43/57 are structurally similar to that of P. vulgaris PrK 44/57 and different from the polysaccharide of P. mirabilis PrK 41/57 studied earlier. The lipopolysaccharides of these strains were tested using enzyme immunosorbent assay, passive hemolysis and Western blot with O-antisera against P. mirabilis 42/57 and P. vulgaris 43/57 and 44/57, as well as with cross-absorbed O-antisera. The chemical and serological data revealed the basis for combining the four strains into Proteus serogroup O23 and division of this serogroup to three subgroups, one for P. vulgaris 43/57 and 44/57 and two others for P. mirabilis 41/57 and 42/57.
5

Classification of Proteus mirabilis TG 115 and CCUG 10701 into the Proteus O23 serogroup based on chemical and serological studies of O-polysaccharides

86%
Zablotni A., Perepelov A. V., Kolodziejska K., Zych K., Knirel Y. A., Sidorczyk Z.
Archivum Immunologiae et Therapiae Experimentalis
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2006
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vol. 54
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issue 6
411-417
EN
Introduction: Bacteria of the genus Proteus are a common cause of urinary tract infections. The O-polysaccharide (OPS) chain of their lipopolysaccharide (LPS) defines the serological specificity of strains. Based on the OPS structures and the immunospecificity of the LPS, Proteus strains have been classified into 74 O-serogroups. Materials and Methods: The OPS of P. mirabilis TG 115 was obtained by mild acid degradation of the LPS and studied by 1H and 13C nuclear magnetic resonance spectroscopy. Antisera were raised by immunization of rabbits with heat-killed bacteria. Serological studies were performed using enzyme immunosorbent assay, passive immunohemolysis, inhibition experiments, absorption of O-antisera, and Western blot.Results: The following structure of the P. mirabilis TG 115 OPS was established: 2)--D-GalpA-(13)--D-GalpNAc-(14)--D-GalpA-(13)--D-GlcpNAc-(1 The same structure has been reported previously for the O-polysaccharides of P. mirabilis CCUG 10701 (O74) and P. mirabilis 41/57 (O23), except that they contain O-acetyl groups in non-stoichiometric quantities. Serological studies showed the antigenic identity of the three strains and their close serological relatedness to P. vulgaris 44/57. Conclusions: Based on the OPS structures and serological data, it is suggested to classify P. mirabilis 41/57, TG 115, and CCUG 10701 into one subgroup and P. mirabilis 42/57 and P. vulgaris 43/57 and 44/57 into another subgroup of the Proteus O23 serogroup.
6

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

86%
Zych K., Kolodziejska K., Drzewiecka D., Perepelov A. V., Knirel Y. A., Sidorczyk Z.
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2007
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vol. 55
|
issue 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.
7

Structures and serology of the O-antigens of Proteus strains classified into serogroup O17 and former serogroup O35

73%
Torzewska A., Grabowski S., Kondakova A. N., Toukach F. V., Senchenkova S. N., Shashkov A. S., Arbatsky N. P., Knirel Y. A., Rozalski A., Kaca W.
Archivum Immunologiae et Therapiae Experimentalis
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2006
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vol. 54
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issue 4
277-282
EN
Introduction: Bacteria of the genus Proteus are facultative pathogens which commonly cause urinary tract infections. Based on the serological specificity of the O-chain polysaccharide of the lipopolysaccharide (O-polysaccharide, O-antigen), strains of P. mirabilis and P. vulgaris have been classified into 60 serogroups. Studies on the chemical structure and serological specificity of the O-antigens aim at the elucidation of the molecular basis and improvement of the serological classification of these bacteria. Materials and Methods: The O-polysaccharide was prepared by acetic acid degradation of the lipopolysaccharide isolated from dried bacterial mass of each strain by hot phenol/water extraction. 1H- and 13C-NMR spectroscopy was used for structural studies. Serological studies were performed with rabbit O-antisera using enzyme immunosorbent assay, passive hemolysis test, and the inhibition of reactions in these assays as well DOC-PAGE and Western blot. Results: Four Proteus strains belonging to serogroups O17 and O35 were found to possess similar O-polysaccharide structures, in particular having the same carbohydrate backbone built up of tetrasaccharide repeating units. However, they differ in the presence or absence of additional substituents, such as phosphoethanolamine in P. mirabilis O17 and glucose in P. penneri O17, as well as in the pattern and degree of O-acetylation of various monosaccharide residues. Serological studies also showed close relationships between the O-antigens studied. Conclusions: Based on these data it is proposed to reclassify strain P. mirabilis PrK 61/57, formerly representing the O35 serogroup, into the serogroup O17 in the Kauffman-Perch classification system of Proteus.
8

Structural and serological characterization of the lipopolysaccharide from proteus penneri 20 and classification of cross-reacting proteus penneri strains 10, 16, 18, 20, 32 and 45 in proteus serogroup O17

73%
Sidorczyk Z., Toukach F. V., Zych K., Arbatsky N. P., Drzewiecka D., Ziolkowski A., Shashkov A. S., Knirel Y. A.
Archivum Immunologiae et Therapiae Experimentalis
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2002
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vol. 50
|
issue 5
345-355
EN
O-specific polysaccharide (O-antigen) of the lipopolysaccharide of Proteus penneri 20 was studied using sugar analysis along with various one- and two-dimensional NMR spectroscopy techniques. The structure of the polysaccharide was established. It has the same carbohydrate backbone structure as that described earlier for P. penneri 16, in which the positions of the O-acetyl groups have not been determined. P. penneri 20 O-antiserum showed a strong cross-reactivity with the lipopolysaccharides of P. penneri 10, 16, 18, 32, 45 and P. mirabilis O17. These data enable classifying these strains together with P. penneri 20 in one Proteus serogroup, O17.
9

Immunochemical studies of the lipopolysaccharides of Hafnia alvei PCM 1219 and other strains with the O-antigens containing D-glucose 1-phosphate and 2-deoxy-2-[(R)-3-hydroxybutyramido]-D-glucose

73%
Katzenellenbogen E., Kocharova N. A., Korzeniowska-Kowal A., Gamian A., Bogulska M., Szostko B., Shashkov A. S., Knirel Y. A.
Archivum Immunologiae et Therapiae Experimentalis
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2008
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vol. 56
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issue 5
347-352
EN
Introduction: Hafnia alvei is the only species of the genus Hafnia, which belongs to the family of Enterobacteriaceae. These Gram-negative bacteria are commonly distributed in the natural environment and are often the cause of human opportunistic infections. Their lipopolysaccharides (LPSs) are important surface antigens which are responsible for the serological specificity and numerous cross-reactions with other enterobacterial genera. So far, 29 different O-polysaccharide (OPS, O-antigen) structures in Hafnia LPSs have been established and for some of them the molecular basis of the serological activity has been elucidated. Materials and Methods: OPS from H. alvei strain PCM 1219 was obtained by mild acid hydrolysis of the LPS followed by gel permeation chromatography of carbohydrate material on Sephadex G-50 column. The polysaccharide structure was determined using chemical methods as well as 13C NMR and 1H NMR spectroscopy. For serological studies, SDS-PAGE, immunoblotting, and passive hemagglutination tests were used. Results: The serological studies revealed a cross-reactivity of the LPSs of H. alvei PCM 1219 and a group of H. alvei strains with an O-antigen containing D-glucose 1-phosphate and [(R)-3-hydroxybutyramido]-D-glucose. The following structure of the OPS was established: 2)-alpha-d-Glcp-(1-PO4-6)-alpha-d-GlcpNAcyl-(14)-alpha-d-GalpNAc-(13)-beta-d-GalpNAc-(1- > 3---- OAc6<-1 alpga-d-Glcp where Acyl stands for (R)-3-hydroxybutyryl and the degree of O-acetylation is 70%. The structure of the core oligosaccharide was found to be typical of the genus Hafnia. Conclusions: Based on the OPS structure and serological results it was concluded that H. alvei strain PCM 1219 should be classified in the same serogroup as the H. alvei type strain ATCC 13337 and five other strains containing D-glucose 1-phosphate and 2-deoxy-2-[(R)-3-hydroxybutyramido]-D-glucose in their O-antigens.
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