The structure of a core oligosaccharide component from Hafnia alvei strain 32 and 1192 lipopolysaccharides.
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Carbohydrate Sequence↗
Biomedical subjects
C Lugowski
Publications and source records attributed to C Lugowski.
Immunochemical characterization of Citrobacter strain PCM 1487 O-specific polysaccharide- and core oligosaccharide-protein conjugates.
O-specific-polysaccharide and core oligosaccharide were isolated from Citrobacter strain PCM 1487 lipopolysaccharide and purified. The polysaccharide was selectively devoid of 4-deoxy-D-arabinohexose residues. Covalent conjugates of the modified O-specific polysaccharide and of the core oligosaccharide with tetanus toxoid were prepared. Immunochemical characterization of these conjugates proved that they are strong immunogens. Using monospecific rabbit antisera raised against the conjugates, the antigenic relationships between lipopolysaccharides of various strains of Citrobacter, Shigella sonnei, and Shigella flexneri were studied by quantitative microprecipitin and quantitative microprecipitin inhibition and immunoblotting tests.
Bacterial Proteins↗
Effect of natural and synthetic immunomodulators on the synthesis of interferon by peritoneal cells of mice.
The effect of different natural and synthetic immunomodulators on the spontaneous interferon (IFN) synthesis by freshly isolated resident peritoneal cells of BALB/c, NZB and C3H mice was investigated. Actinomycetal glycolipids isolated from Curtobacterium betae, Faenia rectivirgula, Rothia dentocariosa and Saccharopolyspora hirsuta at the concentration 1-20 micrograms/ml were found to potentiate the IFN synthesis by the peritoneal cells of BALB/c mice. Similar results were obtained when dsRNA, LPS of Shigella sonnei and lipid A isolated from the LPS were used. The effect of potentiation of the physiological IFN production by the immunomodulators was observed also in the cells of C3H and NZB mice. In contrast, the inhibition of the IFN synthesis was observed when the peritoneal cells of BALB/c and NZB mice were treated with imuthiol at concentration 0.1-10 micrograms/ml. Thymomodulin (TFX-Polfa) at concentration of 1-100 micrograms/ml had no effect on the spontaneous IFN production.
Adjuvants, Immunologic↗
Characterization of an enterobacterial common antigen (ECA) epitope recognized by anti-ECA-tetanus toxoid conjugate serum.
The antigenic reactivity of both native and chemically modified enterobacterial common antigen (ECA) with anti-ECA-tetanus toxoid (TT) conjugate serum was investigated. The results obtained suggest that reduction of the carboxyl group of the mannosaminuronic acid component of ECA diminishes, but does not destroy its antigenic reactivity. Each of the sugar components were found to contribute to its reactivity with anti-ECA-TT conjugate serum, indicating that the trisaccharide repeating unit represents the ECA epitope. A nonasaccharide (trimer of the ECA repeating unit) inhibited antibody binding better than the hexasaccharide dimer, a finding which suggests that oligosaccharide conformation also makes a contribution to its inhibitory activity.
Animals↗
Saccharide-protein covalent conjugates: immunochemical characterization of Citrobacter 036 core oligosaccharide-tetanus toxoid conjugates.
Core oligosaccharides (complete and incomplete) isolated from Citrobacter 036 lipopolysaccharide were covalently conjugated with tetanus toxoid. Serological examination of the Citrobacter 036 core oligosaccharide-tetanus toxoid conjugates showed that they are strong immunogens. The monospecific anti-conjugate sera prepared by immunization of rabbits, were used to study the antigenic relations between lipopolysaccharide core regions of 8 strains of Citrobacter. Immunoelectrophoresis, immunoblotting and quantitative microprecipitation were performed in the experiments.
Animals↗
Structure elucidation of the core regions from Citrobacter O4 and O36 lipopolysaccharides by chemical and enzymatic methods, gas chromatography/mass spectrometry, and NMR spectroscopy at 500 MHz.
Novel enterobacterial core oligosaccharides were isolated from Citrobacter O4 and O36 lipopolysaccharides, and their structures were determined by methylation analysis, Smith degradation and enzymatic degradations, gas chromatography/mass spectrometry, and two-dimensional phase-sensitive correlated, relayed coherence transfer, double-quantum, triple-quantum-filtered, and nuclear Overhauser effect (NOE) 1H NMR spectroscopy at 500 MHz. In the formulas, all hexose residues are D-hexopyranoses, and heptoses are L-glycero-D-manno-heptopyranoses; the alternative locations of the side-chain heptose and pyrophosphorylethanolamine (PPEtN) residues are marked by dashed lines; dOclA stands for 3-deoxy-D-manno-octulosonic acid. (formula; see text) Along with these complete cores, incomplete ones, lacking the hexosamine trisaccharides, occur in the lipopolysaccharides of both types. Qualitative NOE data were in good agreement with the minimum energy conformation of the above O36 oligosaccharide, calculated with the aid of the SUGAR program [Sundin, A., Carter, R. E., & Liljefors, T. (1988) J. Mol. Graphics (in press)].
Acetylglucosaminidase↗
Characterization and diagnostic application of a lipopolysaccharide core oligosaccharide-protein conjugate.
Shigella sonnei phase II core oligosaccharide (OSPhII) was linked covalently with protein (bovine serum albumin or tetanus toxoid) to obtain a conjugate (OSPhII-protein) of good immunogenicity in rabbits. Anti-OSPhII-protein conjugate sera contained high levels of immunoglobulin G antibodies against the complete core region of Sh. sonnei lipopolysaccharide. The antigenic relationships between lipopolysaccharides of R1, R3 and R4 core types using anti-OSPhII-protein sera were studied. These antisera may be used for the rapid determination of core types in unknown enterobacterial strains.
Animals↗
Structural studies of the O-specific side-chains of the Escherichia coli O 10 lipopolysaccharide.
The structure of the O-specific side-chains of the Escherichia coli O 10 lipopolysaccharide has been investigated. Methylation analysis, n.m.r. spectroscopy, and various specific degradations were the principal methods used. It is concluded that the polysaccharide is composed of pentasaccharide repeating-units having the following structure, in which D-Fuc4NAcyl is 4-amino-4,6-dideoxy-D-galactose, acylated with acetic acid (60%) or D-3-hydroxybutyric acid (40%). (Formula: see text).
Carbohydrate Conformation↗
Citrobacter lipopolysaccharides: structure elucidation of the O-specific polysaccharide from strain PCM 1487 by mass spectrometry, one-dimensional and two-dimensional 1H-NMR spectroscopy and methylation analysis.
Structural studies on the O-specific polysaccharide of Citrobacter PCM 1487 lipopolysaccharide, using methylation analysis, Smith degradation and 1H-NMR spectroscopy, indicate that it consists of the trisaccharide repeating units (formula, see text) In this structure, 4-deoxy-D-araHex stands for 4-deoxy-D-arabino-hexose.
Carbohydrate Conformation↗
The enterobacterial common-antigen, a cyclic polysaccharide.
Structural studies of the enterobacterial common-antigen, using chemical methods and fast-atom-bombardment mass spectrometry, indicate that it is a cyclic polysaccharide, composed of four, five, and, to a smaller extent, six trisaccharide repeating-units. In the structure of the antigen, given below, D-Fuc4NAc stands for 4-acetamido-4,6-dideoxy-D-galactose.
Antigens, Bacterial↗
Structural determination of the capsular polysaccharide of Streptococcus pneumoniae type 18C (56).
The specific capsular polysaccharide of Streptococcus pneumoniae type 18C (56) contains D-glucose, D-galactose, L-rhamnose, and glycerol residues, and phosphate and O-acetyl groups in the molar ratios of 3:1:1:1:1:1. Accumulated data from methylation analyses of the native and the specifically degraded, native polysaccharide indicated that it is composed of the repeating unit shown; it also contains O-acetyl groups, of undetermined location, in the molar ratio to L-rhamnose of 1:1. (formula; see text).
Carbohydrate Conformation↗
Conjugation of meningococcal lipopolysaccharide R-type oligosaccharides to tetanus toxoid as route to a potential vaccine against group B Neisseria meningitidis.
Oligosaccharides were obtained by the mild acid hydrolysis of the lipopolysaccharides from a number of different strains of Neisseria meningitidis, serotypes L2, L3, L4, L5, and L10. The dephosphorylated oligosaccharides were conjugated to tetanus toxoid as their 2-(4-isothiocyanatophenyl)-ethylamine derivatives, which resulted in the incorporation of from 18 to 38 oligosaccharides per molecule of tetanus toxoid. When injected in rabbits, the conjugates produced oligosaccharide-specific antibodies which were predominantly serologically specific but which also exhibited some cross-reactivity. These serological results can be attributed to regions of structural dissimilarity and similarity within the oligosaccharides. The oligosaccharide-specific antibodies were also lipopolysaccharide serotype specific, thus indicating that the oligosaccharides are the determinants associated with this serotype specificity. Consistent with the serological results, the conjugate antisera were bactericidal for the homologous serotype meningococcal organisms and in some cases for heterologous serotype organisms.
Animals↗
Characterization of anti-Citrobacter 036 specific polysaccharide monoclonal antibodies.
Monoclonal mouse antibodies specific for the 0 antigen of Citrobacter 036, a homopolymer of beta (1----2)-linked 4-deoxy-D-arabinohexose, were generated by the hybridoma technique. Balb/c mice were immunized with killed whole-cell vaccine and initial selection of active clones was based on enzyme-linked immunosorbent assay (ELISA) employing purified lipopolysaccharide (LPS). Concentrated culture supernatants from selected hybrid cultures were used to identify 10 0-antigen specific monoclonal antibodies using the multiple criteria of immunoprecipitation of 0 chains and LPS, inhibition by acid hydrolyzed 0 chains in the screening ELISA, and antibody class analysis. Four monoclonal antibodies were chosen for further study using dose-dependent 0-chain inhibition of ELISA and passive hemagglutination, passive hemolysis, and bacterial agglutination titres. When screened with Citrobacter serotypes known to contain the sugar 4-deoxy-D-arabinose, passive hemagglutination tests showed that the two monoclonal antibodies examined possessed titres which could be correlated with the reported 4-deoxy-D-arabinohexose content of the respective LPS's. This sugar is an antigenically important unit of several Citrobacter serotypes as defined by these well-characterized monoclonal antibodies.
Animals↗
Structure of native polysaccharide antigens of type Ia and type Ib group B Streptococcus.
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Antigens, Bacterial↗
Enterobacterial common antigen-tetanus toxoid conjugate as immunogen.
The methods of limited periodate oxidation and reductive amination were used to obtain covalently linked enterobacterial common antigen (ECA) with tetanus toxoid. This procedure is simple and gives a good yield of the conjugate with high ECA content (molecular ratio of ECA to tetanus toxoid, 6:1). The ECA-tetanus toxoid conjugate is immunogenic in rabbits, in contrast to free ECA or a mixture of ECA with proteins. This conjugate produces high levels of ECA-specific immunoglobulin G antibodies, which can be used as a standard serum.
Animals↗
Interaction of Wratizolin with serum albumins and its antiproteolytic activity.
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Animals↗
Isolation of a serogroup 1-specific antigen from Legionella pneumophila.
Outer membrane material was extracted from a serogroup 1 strain of Legionella pneumophila using ethylenediaminetetraacetate. Ferritin-conjugated antiserum reacted only at the surface of the organism, as seen by electron microscopy. Outer membrane material was fractionated into five peaks by molecular sieve chromatography using a gel that had been equilibrated in a buffer containing sodium deoxycholate. One resolved peak (approximately 4x10(4) daltons) was highly active serologically. When rechromatographed in the absence of sodium deoxycholate, material from this peak reaggregated to approximately 10(6) daltons. Ther serologic activity of this antigen was restricted to L. pneumophila serogroup 2, although minor cross-reactions with strains of serogroups 2 and 4 were detected using an enzyme-linked immunosorbent assay. The antigen was less than 10% carbohydrate, 15% protein, 1.1% phosphate, and the remainder lipid of unknown composition. Neither 2-keto-3-deoxyoctonate nor heptose was detected, and the antigen did not induce a Shwartzman reaction. Only one band was seen on sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Animals↗
Conformational aspects critical to the immunospecificity of the type III group B streptococcal polysaccharide.
Immunization of rabbits with group B type III streptococcus organisms induces two distinct populations of antibodies with a specificity for determinants on the native capsular polysaccharide antigen of these organisms. Some of the structural and conformational features of the two determinants responsible for the formation of these antibodies were elucidated by (13)C NMR and serological studies on the native type III polysaccharide and some of its structurally modified analogues. The specificity of the determinant corresponding to the major population of antibodies is dependent of the presence of sialic acid residues on the native type III antigen, and although these residues are not an integral part of the determinant, they exert conformational control over it. The carboxylate groups of the sialic acid residues are an important factor in this control mechanism which could possibly involve intramolecular hydrogen bonding. The terminal sialic acid residues control the orientation of the penultimate beta-d-galactopyranose residues with respect to the backbone of the native antigen. The orientation of these residues is critical to the determinant because the determinant is probably small and is located precisely at the junction of the same beta-d-galactopyranose residues with the backbone of the native type III antigen. The determinant corresponding to the other population of antibodies is not sialic acid dependent. This determinant is located on the backbone of the native antigen in the vicinity of the other determinant but on the opposite side to the oligosaccharide branches. In this position, its conformation is unaffected even by the removal of the oligosaccharide branches from the native antigen.
Antibodies↗