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PubMed1988

[Antigenic polysaccharides of bacteria. 34. Structure of O-specific polysaccharide chains of lipopolysaccharides from Pseudomonas cepacia strains IMV 4207 (Serotype A) and IMV 598/2].

On the basis of non-destructive analysis by means of 1H and 13C NMR spectroscopy and calculation of specific optical rotation, it was concluded that O-specific polysaccharide of Pseudomonas cepacia strain IMV 4207 (serotype A) has the structure (I): (formula; see text) Two structurally different polysaccharides were found in the ratio of approximately 2.5:1 in P. cepacia strain IMV 598/2 which is serologically related to serotype A in Nakamura classification and serotype 2 in Heidt classification. The minor polysaccharide has the structure (I) whereas the major one possesses the structure (II) which is characteristic of the formerly studied O-specific polysaccharide of P. cepacia strain IMV 4137 belonging to serotype 2: ----4)-beta-D-Galp-(1----2)-alpha-L-Rhap-(1----.

Antigens, Bacterial↗
PubMed1965

THE BIOSYNTHESIS OF POLYSACCHARIDES. INCORPORATION OF D-(1-14C)GLUCOSE AND D-(6-14C)GLUCOSE INTO PLUM-LEAF POLYSACCHARIDES.

1. The utilization of specifically labelled d-glucose in the biosynthesis of plum-leaf polysaccharides has been studied. After these precursors had been metabolized in plum leaves, the polysaccharides were isolated from the leaves, and their monosaccharide constituents isolated and purified. 2. Both the specific activities and the distribution of (14)C along the carbon chains of the monosaccharides were determined. Significant (14)C activity was found in units of d-galactose, d-glucose, d-xylose and l-arabinose, but their specific activities varied widely. The labelling patterns suggest that in the leaves the other monosaccharides all arise directly from d-glucose without any skeletal change in the carbon chain, other than the loss of a terminal carbon atom in the synthesis of pentoses. 3. The results indicated that within the leaf there are various precursor pools for polysaccharide synthesis and that these pools are not in equilibrium with one another.

Arabinose↗
PubMed1987

The role of capsular polysaccharide K21b of Klebsiella and of the structurally related colanic-acid polysaccharide of Escherichia coli in resistance to phagocytosis and serum killing.

The behaviour of strains of Klebsiella aerogenes of capsular serotype K21 and strains of Escherichia coli producing a structurally related polysaccharide (colanic acid) was analysed by phagocytic and serum-killing assays. The cell-surface characteristics of these strains and of non-capsulate strains derived from them were also investigated by partitioning experiments in aqueous two-polymer phase systems. The possession of K21-type capsule by K. aerogenes or colanic-acid polysaccharide by E. coli conferred a strong negative charge on capsulate bacteria. Negatively charged bacteria of E. coli producing colanic-acid capsules, however, like non-capsulate K. aerogenes, were susceptible to uptake by polymorphonuclear leukocytes. In contrast, K21 polysaccharide conferred on klebsiellae considerable resistance to phagocytic uptake. The finding that ingested non-capsulate derivative strains of K. aerogenes were less rapidly degraded by phagocytes than E. coli strains suggested that other components of the cell surface of Klebsiella, notably lipopolysaccharide, may be involved in protection against phagocytic killing. The presence of colanic-acid capsules on E. coli conferred little resistance to the bactericidal activity of human serum or phagocytic uptake and did not protect against intracellular killing by polymorphonuclear leukocytes.

Blood Bactericidal Activity↗
PubMed1963

Polysaccharide produced by the genus Pullularia. I. Production of polysaccharide by growing cells.

A dextranlike polysaccharide was found to be produced on substrates of sucrose, maltose, glucose, and fructose by growing cells of various strains of the genus Pullularia. The polysaccharide, obtainable in amounts as large as 2 to 3 g per 100 ml of culture medium using various carbohydrates as the carbon source, was soluble in cold water but not in 50% alcohol. The polysaccharide obtained had a alpha(d) = +197.5 degrees (c = 0.2 in water), and its molecular weight, determined by the light-scattering method, was found to be approximately 250,000.

Carbohydrates↗
PubMed1996

[Study on the enhancing effect of polyporus polysaccharide, mycobacterium polysaccharide and lentinan on lymphokine-activated killer cell activity in vitro].

The actions of Polyporus polysaccharide (PPS), mycobacterium polysaccharide (MPS) and lentinan (LEN) on lymphokine-activated killer (LAK) cell activity in vitro were investigated in this study. Human peripheral blood mononuclear cells (PBMC) were cultured for 96 hours with medium containing different concentrations of the above-mentioned drugs in combination with recombinant interleukin 2 (rIL-2). Then cell-mediated lysis was determined by 1H-TdR release assay including NK sensitive and NK resistant target cells. The results demonstrated that, when combined with rIL-2 in a certain concentration, all three kinds of polysaccharides could enhance the LAK activity by 42%-56.9%, and reduce the dose of rIL-2 by 50% (P < 0.05-0.01). It suggested that the PPS, MPS and LEN could be used as bioactivity regulators in LAK cell therapy in tumor treatment.

Cytotoxicity, Immunologic↗
PubMed2006

Polysaccharide microarrays for polysaccharide-platelet-derived-growth-factor interaction studies.

Polysaccharide microarrays have great potential for the high-throughput analysis of polysaccharide-protein interactions. Here we demonstrate that a polysaccharide microarray prepared by printing a library of dextran polymers derivatized by methylcarboxylate, benzylamide, and sulfate groups (DMCBSu compounds) on to glass slides permitted the rapid identification of a set of compounds able to interact with the platelet-derived growth factor BB (PDGF-BB) isoform, a growth factor involved in wound healing. Microarray interaction results were compared to the capacity of DMCBSu compounds to potentiate the in vitro PDGF-BB-induced proliferation of human dermal fibroblasts.

Becaplermin↗
PubMed1992

Induction of anti-pneumococcal cell wall polysaccharide antibodies by type 4 pneumococcal polysaccharide-protein conjugates.

We have prepared polysaccharide-protein conjugates consisting of type 4 pneumococcal capsular polysaccharide (PS4) coupled to tetanus toxoid. The PS4 preparation used contained 2.5% pneumococcal cell wall polysaccharide (CPs). During the conjugation process, in addition to PS4-protein conjugates, CPs-protein conjugates were also formed. After immunization with PS4-protein conjugates, CPs-protein conjugates that are present as a contaminant induce IgG anti-CPs antibodies in mice. Pneumococcal oligosaccharides, prepared by periodate oxidation of PS4, did not contain detectable amounts of CPs; hence, oligosaccharide-protein conjugates did not induce anti-CPs antibodies.

Animals↗
PubMed1985

Intra-axonal polysaccharide deposits in the peripheral nerve seen in adult polysaccharide storage myopathy.

This is the first report of an adult polysaccharide storage myopathy demonstrating polysaccharide deposits within the axons of the peripheral nerve. Histochemical and electron-microscopic studies revealed that the intra-axonal deposits were basophilic, PAS-positive, diastase-resistant, and composed of filamentous and granular material. The structural similarity among the polysaccharide deposits in our case, amylopectin-like materials in type IV glycogenosis, Lafora bodies, corpora amylacea, and polyglucosan bodies is discussed.

Adult↗
PubMed2006

Complement activation by core-shell poly(isobutylcyanoacrylate)-polysaccharide nanoparticles: influences of surface morphology, length, and type of polysaccharide.

PURPOSE: Biodistribution of intravenously administered nanoparticles depends on opsonization. The aim of this study was the evaluation of complement activation induced by nanoparticles coated with different polysaccharides. Influences of size and configuration of dextran, dextran sulfate, or chitosan bound onto nanoparticles were investigated. METHOD: Core-shell nanoparticles were prepared by redox radical or anionic polymerization of isobutylcyanoacrylate in the presence of polysaccharides. Conversion of C3 into C3b in serum incubated with nanoparticles was evaluated. RESULTS: Cleavage of C3 increased with size of dextran bound in "loops" configuration, whereas it decreased when dextran was bound in "brush." It was explained by an increasing steric repulsive effect of the brush, inducing poor accessibility to OH groups. The same trend was observed for chitosan-coated nanoparticles. Nanoparticles coated with a brush of chitosan activated the complement system lesser than nanoparticles coated with a brush of dextran. This was explained by an improved repelling effect. Dextran-sulfate-coated nanoparticles induced a low cleavage of C3 whereas it strongly enhanced protein adsorption. CONCLUSION: Complement activation was highly sensitive to surface features of the nanoparticles. Type of polysaccharide, configuration on the surface, and accessibility to reactive functions along chains are critical parameters for complement activation.

Chitosan↗
PubMed1987

New n.m.r.-spectroscopic approaches for structural studies of polysaccharides: application to the Haemophilus influenzae type a capsular polysaccharide.

The extension of several modern nuclear magnetic resonance (n.m.r.) spectroscopic techniques to polysaccharides is discussed and illustrated, using the native Haemophilus influenzae type a capsular polysaccharide. These techniques provide for the unambiguous assignment of all n.m.r. resonances (1H, 13C, and 31P) via high-sensitivity homonuclear and 1H-detected heteronuclear correlations, and they are capable of locating the intersaccharide linkages (both O-linked and phosphoric diester-linked) and appended groups (e.g. O-acetyl groups). To illustrate the power and sensitivity of these methods, a 10-mg sample of the H. Influenzae type a polysaccharide (repeat unit mol. wt. = 376) was studied. The combined acquisition time for the two-dimensional 1H-13C correlation data (one-bond and multiple-bond), the 1H-31P correlation data, and the 1H-1H (homonuclear Hartmann-Hahn) data was approximately 18 h.

Carbohydrate Conformation↗
PubMed1983

Influences of anionic polysaccharides on DNA synthesis in isolated nuclei and by DNA polymerase alpha: correlation of observed effects with properties of the polysaccharides.

The basis of the differential effect of anionic polysaccharides on replicative DNA synthesis in liver and hepatoma cell nuclei was investigated. The differential effect of heparin was lost when more than 40% of its sulfate was removed. DNA synthesis in liver nuclei was optimally stimulated by heparin of molecular weight 22600 and sulfate to hexosamine ratio 2.42, but inhibited by heparin of molecular weight 4300 and sulfate to hexosamine ratio 2.35. A heparin fragment (molecular weight 2800 and sulfate to hexosamine ratio 1.81), prepared by partial nitrous acid treatment was a potent inhibitor of DNA synthesis in hepatoma nuclei. There was no significant difference in the rate of entry of heparin or its subfractions into either liver or hepatoma nuclei. In both cases less than 15% of added polysaccharide entered the nuclei and only about 4.5% was found associated with the chromatin. The influence of the anionic polysaccharides on DNA synthesis was correlated with their ability to complex with histones as determined by relative light scattering in a laser nephelometer. The relative light scattered on mixing with histones (H1, H2A + H3, H4) was high for DNA synthesis stimulators (heparin, dextran sulfate); medium for DNA synthesis inhibitors (chondroitin 4- and 6-sulfates, heparan sulfate) and low for non-effectors (keratan sulfate, hyaluronic acid). Heparin and chondroitin sulfate H, which at low concentrations stimulate DNA synthesis in liver nuclei, inhibited DNA synthesis by calf thymus DNA polymerase alpha at all concentrations. This inhibition was not simply due to electrostatic interactions.

Animals↗
PubMed1992

Structure of the Escherichia coli 0104 polysaccharide and its identity with the capsular K9 polysaccharide.

The 0104 antigen (lipopolysaccharide, LPS) of Escherichia coli has an acidic O specific polysaccharide. From the aqueous phase of a phenol water extraction of E. coli O104: K-, a fraction was obtained by ultracentrifugation and Cetavlon precipitation of the supernatant, which was enriched in long-chain LPS. Compositional analysis, NMR spectroscopy, periodate oxidation and methylation analysis showed that the polysaccharide chain of O104 LPS II consisted of galactose, N-acetylgalactosamine and neuraminic acid and acetate in the molar ratio of 2:1:1:1 and contained 3-beta Gal, 3-beta GalNAc, 4-alpha Gal, and 4-alpha(9-OAc-NeuNAc) in linear sequence. The same results were obtained with the capsular K9 polysaccharide from E. coli O9:K9, as presented here and reported previously (Dutton et al. (1987) Carbohydr. Res. 170, 193-206).

Bacterial Capsules↗
PubMed2003

Induction of a protective capsular polysaccharide antibody response to a multiepitope DNA vaccine encoding a peptide mimic of meningococcal serogroup C capsular polysaccharide.

Systemic infection by encapsulated organisms, such as Neisseria meningitidis, is a major cause of morbidity and mortality worldwide, especially in individuals less than 2 years of age. Antibodies directed at the capsular polysaccharide are shown to be protective against disease by inducing complement-dependent bactericidal activity. The current polysaccharide vaccine has been shown to be poorly immunogenic in high-risk groups and this is probably related to its T-independent properties. An alternative approach to eliciting a T-dependent serum immunoglobulin G (IgG) antibody response to encapsulated pathogens is DNA vaccination. We assessed the immunogenicity of a multiepitope DNA vaccine encoding a T-cell helper epitope and a peptide mimic of N. meningitidis serogroup C. The DNA construct induced a significant anti-polysaccharide antibody response that was bactericidal. Mice immunized with the DNA construct were subsequently protected against challenge with a lethal dose of N. meningitidis serogroup C.

Animals↗
PubMed1981

Further studies of the polysaccharide of Klebsiella pneumoniae possessing strong adjuvanticity. II. Serological relationship between the adjuvant polysaccharide and O3 antigen of Klebsiella.

The serological specificity of the neutral polysaccharide possessing extraordinarily strong adjuvanticity originally isolated from the culture supernatant of Klebsiella K1 strain Kasuya has been investigated. Among all of the reference strains (K1-K82) of Klebsiella obtained from the International Escherichia and Klebsiella Center, Statens Seruminstitut, Copenhagen, only 13 strains have been shown to produce the adjuvant polysaccharide by the passive hemagglutination inhibition test. All of these 13 strains belong to the O3 group, and the strains which belong to other O groups of which were not identifiable did not produce it. The gel precipitation test has demonstrated that the adjuvant polysaccharide is antigenically identical to O3 antigen isolated from the cells of the decapsulated mutant (strain LEN 1) of Klebsiella K1 strain Kasuya and to O9 antigen of Escherichia coli isolated from either the culture supernatant or the cells, which has already been shown to be antigenically and structurally identical to the O3 antigen of Klebsiella.

Adjuvants, Immunologic↗
PubMed1993

Structural determination of the capsular polysaccharide produced by Klebsiella pneumoniae serotype K40. NMR studies of the oligosaccharide obtained upon depolymerisation of the polysaccharide with a bacteriophage-associated endoglycanase.

The Klebsiella pneumoniae K40 capsular polysaccharide has been isolated and investigated by use of methylation analysis, specific degradations and NMR spectroscopy. The polysaccharide was depolymerised by a bacteriophage-associated endogalactosidase, and the resulting oligosaccharide was characterised by one-dimensional and two-dimensional NMR spectroscopy and direct chemical ionisation MS. The repeating unit of the K40 capsular polysaccharide was shown to be a linear hexasaccharide with the composition-->3)- alpha-L-Rhap-(1-->2)-alpha-L-Rhap-(1-->4)-alpha-D-GlcpA++ +-(1-->2-)- alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->3)-alpha-D-Galp-(1--> (Rha, rhamnose).

Bacteriophages↗
PubMed1997

Identification of the common antigenic determinant shared by Streptococcus pneumoniae serotypes 35A and 20 capsular polysaccharides--structural analysis of the Streptococcus pneumoniae serotype 35A capsular polysaccharide.

The specific polysaccharide antigen of Streptococcus pneumoniae serotype 35A was shown, by a combination of one- and two-dimensional NMR methods and chemical analyses, to be a high-molecular-mass polymer composed of D-galactose, D-glucose, mannitol, and phosphate (3:1:1:1). The pentasaccharide repeating unit is polymerized through phosphate diester linkages to give the structure, [formula in text] O-Acetyl substituents are present at positions 5 and 6 of the 3)-beta-D-Galf residue and at position 2 of the 6)-beta-D-Galf residue. The capsular polysaccharides of S. pneumoniae serotypes 20 and 35A both contain the disaccharide unit -->3)-beta-D-Galf-(1-->3)-beta-D-Glcp-(1--> which is the probable structural determinant responsible for the serological cross reactivity of the two polysaccharides.

Bacterial Capsules↗
PubMed1983

Purification and immunochemical properties of Escherichia coli B polysaccharide cross-reacting with Salmonella typhi Vi antigen: preliminary evidence for cross-reaction of the polysaccharide with Escherichia coli K1 antigen.

An acidic polysaccharide of Escherichia coli B was isolated by a mild procedure and purified to homogeneity. The polysaccharide was found to react in Salmonella typhi Vi antisera and E. coli K1 antisera. Serological analysis and preliminary chemical characterization of the polysaccharide indicated that it is an aminouronic acid polymer which, although not structurally identical to either Vi or K1, appears more like the Vi antigen, both immunochemically and chemically.

Antigens↗
PubMed1985

Immunity and protection of mice against Neisseria meningitidis group B by vaccination, using polysaccharide complexed with outer membrane proteins: a comparison with purified B polysaccharide.

A methodology for preparing outer membrane proteins (type specific) complexed to group B polysaccharide of Neisseria meningitidis is described. These complexes, low in nucleic acid and lipopolysaccharide content, were immunogenic in mice with induction of humoral antigroup B and antiprotein responses. Immunized mice were also protected against challenge with N. meningitidis group B strains of the same or a different type from that used for vaccination. Both immunity and protection were enhanced when the mice received a secondary immunization with the protein-polysaccharide complex. Additional data have shown the capacity of purified B polysaccharide to induce immunological memory, even though it is incapable of inducing a humoral response when given alone.

Animals↗