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PubMed1984

Serum antibodies after vaccination with Haemophilus influenzae type b capsular polysaccharide and responses to reimmunization: no evidence of immunologic tolerance or memory.

Haemophilus influenzae type b capsular polysaccharide vaccine was given in Finland in 1974 to approximately 50,000 infants and children, whose serum anti-H influenzae type b capsular polysaccharide levels have been followed for 3 1/2 years. The serum antibodies induced by the vaccination proved short-lived (less than 6 months) in the infants younger than 18 months. Elevated serum antibody levels were detectable for 1 1/2 years but less than 3 1/2 years in the children who were vaccinated when 18 to 35 months old. In the children who were 3 to 5 years old when vaccinated, the elevated anti-H influenzae type b capsular polysaccharide levels persisted for at least 3 1/2 years. Therefore, children vaccinated at the age of 18 months may need a new dose of vaccine 1 to 1 1/2 years after the first dose in order to be protected for the period of high susceptibility, until the age of approximately 7 years. Some of the vaccinated children were reimmunized 3 1/2 years after the first dose, and the anti-H influenzae type b capsular polysaccharide levels in their sera were studied in a similar manner. At no time did the anti-H influenzae type b capsular polysaccharide levels after the reimmunization differ from the anti-H influenzae type b capsular polysaccharide levels seen after the first vaccination in children of the same age.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

PubMed1981

A simple method for the extraction of polysaccharide B from Brucella cells for use in the radial immunodiffusion test diagnosis of bovine brucellosis.

In order to extract the polysaccharide B of brucella used in Radial Immunodiffusion test (RID) for detection of infected cattle, a quick, simple and safe method is proposed. It consists of using phenol killed cells of B. melitensis 16 M known to produce polysaccharide B. After autoclaving brucella cells resuspended in saline, the cells are separated by centrifugation and the polysaccharide B is obtained by two successive ethanol precipitations of the supernatant. Comparison of the results of RID tests carried out with the polysaccharide extract of B. melitensis 16 M (rapid method) and 231 sera of infected cattle shows the validity of this method of preparation. Differences in yield (1.5% for the second precipitate against 0.5% for the polysaccharide-reference and in optimum dose used the reaction (10 microgram against 200 microgram) show that as many as 60 times more RID tests can be made with the polysaccharide obtained by rapid method.

Animals↗

PubMed1993

[Immunological principles of polysaccharide-protein conjugate vaccination].

Haemophilus influenzae and Streptococcus pneumoniae are bacteria with a polysaccharide capsule. The production of specific antibodies against capsular polysaccharide plays a pivotal role in the defence against these organisms. However, children under the age of two years do not at all or only poorly respond to an infection with encapsulated bacteria or after vaccination with purified capsular polysaccharide antigen. In contrast, protein antigens, e.g. tetanus- and diphtheriae-toxoid, are good immunogens in this age group. The difference between polysaccharide antigen and protein antigen is that the former are T-dependent antigens whereas the latter are T-independent. The reason for this age dependent "immunodeficiency" is not clear. A functional immaturity of a B-cell population seems to be the reason for the unresponsiveness of young children against Ti-2 antigens. The Haemophilus conjugate vaccine contains the capsular polysaccharide chemically conjugated to a carrier protein. This results in an immune response against the polysaccharide with characteristics of a T dependent antigen, giving high immunogenicity even in children under the age of two years, resulting in high antibody-production and the induction of immunological memory.

Antibodies, Bacterial↗

PubMedDate not supplied

Capsular polysaccharide serotypes of coagulase-positive staphylococci associated with tenosynovitis, osteomyelitis, and other invasive infections in chickens and turkeys: evidence for new capsular types.

To study the role of capsular polysaccharides in staphylococcal infections in chickens and turkeys, we surveyed 101 coagulase-positive isolates obtained from investigators in geographically diverse locales in the United States and Canada. Fifty-six isolates were obtained from chickens; 51 (91%) were type 5, and 5 (9%) were type 8. Forty isolates were obtained from turkeys; 13 (33%) were type 5, 15 (38%) were type 8, and 12 (29%) were untypable. Five type 5 isolates were obtained from poultry of uncertain species. Two untypable turkey isolates were studied further. Rabbit serum obtained after immunization with formalin-killed staphylococci was type-specific as assessed by a tube agglutination test and promoted opsonophagocytosis of the isolate that had been used for immunization. Chromatography of carboxy-reduced capsular polysaccharide from one of the isolates suggested that mannoseaminouronic acid was present in the unreduced polysaccharide. The spectrum obtained by 13C-nuclear magnetic resonance suggested the presence of N-acetyl sugars such as mannoseaminouronic acid and 6-deoxy sugars such as N-acetyl fucosamine. The structure of the polysaccharide differed from that of the type 5 and type 8 capsular polysaccharides. Thus, poultry isolates can be classified into at least four serotypes on the basis of their capsular polysaccharides. This observation provides a new focus for investigation into the epidemiology and virulence determinants of this important pathogen in chickens and turkeys.

Animals↗

PubMed1998

[Polysaccharides of flower plants: structure and physiological activity].

The results of studies on the chemical structure and physiological activity of phanerogam polysaccharides, accumulated within the last two decades, are reviewed. Three types of polysaccharides are considered: rhamnogalacturonans (pectins and related gums and mucilages, type A), acidic arabinogalactans (mainly plant mucilages, gums, and some hemicelluloses, type B), and neutral glucans and heteroglycans (reserve polysaccharides, type C). Various physiological activities of these plant polysaccharides are discussed, with particular emphasis being placed on their immunomodulatory action. The data available on the relationship between chemical structure and physiological activity of plant polysaccharides are considered. Information on the medicinal use of some plants containing physiologically active polysaccharides is presented.

Adjuvants, Immunologic↗

PubMed1986

Polysaccharide lyases.

Polysaccharide lyases (or eliminases) are a class of enzymes (EC 4.2.2.-) that act to cleave certain activated glycosidic linkages present in acidic polysaccharides. These enzymes act through an eliminase mechanism, rather than through hydrolysis, resulting in unsaturated oligosaccharide products. Acidic polysaccharides are ubiquitous and so are the lyases that degrade them. This review article examines lyases that act on acidic polysaccharides of plant, animal, and microbial origin. These lyases are predominantly of microbial origin and come from a wide variety of both pathogenic and nonpathogenic bacteria and fungi. The lyases discussed include alginate lyase (EC 4.2.2.3), pectin lyase (EC 4.2.2.10), pectate lyase (EC 4.2.2.2), oligogalacturonide lyase (EC 4.2.2.6), exopolygalacturonate lyase (EC 4.2.2.9), chondroitin lyases (EC 4.2.2.4 and EC 4.2.2.5), hyaluronate lyase (EC 4.2.2.1), heparin lyase (EC 4.2.2.7), heparan lyase (EC 4.2.2.8), and other unclassified lyases. This review examines the sources, regulation, purification, and properties of these polysaccharide lyases.

Animals↗

PubMed1999

Antiviral Activities of Marine Pseudomonas Polysaccharides and Their Oversulfated Derivatives.

: A marine Pseudomonas species WAK-1 strain simultaneously produces extracellular glycosaminoglycan and sulfated polysaccharide. Among the antiviral activities tested for these polysaccharides, the latter showed anti-HSV-1 activity in RPMI 8226 cells (50% effective concentration is 1.4 µg/ml). Oversulfated derivatives of these polysaccharides prepared by dicyclohexylcarbodiimide-mediated reaction for both polysaccharides showed antiviral activities against influenza virus type A (for glycosaminoglycan, 50% effective concentration is 11.0 µg/ml; for another, 2.9 µg/ml). Glycosaminoglycan, sulfated polysaccharide, and their chemically synthesized oversulfated derivatives did not show antiviral activities against influenza virus type B and human immunodeficiency virus type 1. No cytotoxicity of these products was noted against host cells at the 50% cytotoxic concentration of 100 µg/ml, except that naturally occurring sulfated polysaccharide had 50% cytotoxicity against MT-4 cells at 8-21 µg/ml.

Journal Article↗

PubMed1991

Heterogeneity and characterisation of mitogenic and anti-complementary pectic polysaccharides from the roots of Glycyrrhiza uralensis Fisch et D.C.

Two anti-complementary polysaccharide fractions (GR-2IIa and GR-2IIb), isolated from the roots of Glycyrrhiza uralensis Fisch et D.C., each gave five anti-complementary polysaccharides (GR-2IIa-1-5 and GR-2IIb-1-5) on h.p.l.c.; likewise, GR-2IIc gave two anti-complementary and mitogenic polysaccharides (GR-2IIc-1-2A and -2IIc-2) by gel filtration and h.p.l.c. GR-2IIc-1-2A showed the most potent anti-complementary activity. GR-2IIa-1-5 and GR-2IIb-1-5 contained 40-85% and 50-90% of GalA, respectively, in addition to Rha, Ara, and Gal. GR-2IIc-1-2A and -2IIc-2 mainly comprised Glc, Gal, GalA, and GlcA in addition to Rha, Fuc, Xyl, Ara, and Man. Methylation analysis and digestion with endo-alpha-(1----4)-polygalacturonase indicated that all of the polysaccharides contained polygalacturonan regions which were frequently methyl-esterified. GR-2II-a, -2IIb, and -2IIc gave enzyme-resistant fractions of large and intermediate sizes, in addition to oligogalacturonides. Each large fraction from GR-2IIa and -2IIb consisted mainly of Ara, Gal, and GalA, whereas the intermediate fractions were composed of small proportions of 2-Me-Fuc, 2-Me-Xyl, and apiose (Api), in addition to Rha, Ara, Gal, and GalA. The large fraction from GR-2IIc mainly contained Rha, Man, Gal, and GalA in addition to Fuc, Ara, Xyl, and Glc, whereas the intermediate fraction consisted of 2-Me-Fuc, 2-Me-Xyl, and Api, in addition to Rha, Ara, GalA, Fuc, Xyl, Man, Gal, and Glc. Base-catalysed beta-elimination followed by ethylation indicated that all the polysaccharides except GR-2IIc-2 contained a 4-linked uronic acid attached to position 2 of 2,4-linked Rha. Single radial gel diffusion, using the beta-D-glucosyl-Yariv antigen, indicated that GR-2IIa-1 and GR-2IIc-2 contained relatively large proportions of (1----3,6)-beta-D-galactan moieties. The anti-complementary activities of GR-2IIa-3, GR-2IIa-4, and GR-2IIb-4 decreased after de-esterification followed by digestion with endo-alpha-(1----4)-polygalacturonase. The large fractions from GR-2IIa-2IIc showed more potent anti-complementary activities than the original polysaccharide fractions, whereas the intermediate fractions and oligogalacturonides were inactive. The large fraction from GR-2IIc had more potent mitogenic activity than GR-2IIc, whereas the intermediate fraction and oligogalacturonides from GR-2IIc were inactive.

Adjuvants, Immunologic↗

PubMed1985

Lymphocytes express a diverse array of specific receptors for sulfated polysaccharides.

Lymphocyte receptors for sulfated polysaccharides were detected in two ways, namely, by the ability of lymphocytes to form rosettes with sheep red blood cells (SRBC) coupled with one of fourteen different sulfated polysaccharides, and by the ability of cholate extracts of lymphocytes to hemagglutinate the same sulfated polysaccharide-coupled SRBC. It was found that murine lymphocytes lacked receptors for a number of glycosaminoglycans, such as hyaluronic acid, chondroitin-4-sulfate, chondroitin-6-sulfate, and dermatan sulfate, but reacted strongly with heparin, arteparon, and a number of sulfated polysaccharides of plant and bacterial origin. In each case receptor activity was demonstrated by rosetting and by the ability of lymphocyte lysates to strongly agglutinate sulfated polysaccharide-coupled SRBC. The receptors exhibited a high degree of diversity as evidenced by (a) only subpopulations of lymphocytes, particularly splenic B cells, expressing receptors for some of the sulfated polysaccharides and (b) hemagglutination-inhibition analyses revealing numerous subsets of receptors with different binding specificities. Receptor diversity was further highlighted by a 48% difference in the hemagglutination-inhibiton results between thymus and spleen. It is proposed that these receptors are involved in cell-cell communication and lymphocyte homing and recirculation. The likely target structures for the receptors in vivo are the heparan sulfates, a ubiquitous and structurally diverse family of sulfated glycosaminoglycans.

Animals↗

PubMed2006

The presence of natural human antibodies reactive against pharmacologically active pectic polysaccharides from herbal medicines.

Direct ELISA was performed using normal human sera and human colostrum, to analyse the presence of antibodies which react with pharmacologically active pectic polysaccharides isolated from plants used in traditional Japanese herbal (Kampo) medicine. All sera and colostrum were shown to contain IgM, IgG, IgA and secretory IgA class antibodies which react with the active pectic polysaccharides to different degrees. The reacting IgG antibody in normal human serum recognized the ramified regions (rhamnogalacturonan core with carbohydrate side-chains) of the pharmacologically active pectic polysaccharides as the active sites for complement-activating activity. Correlation analysis indicated that a significant and positive correlation was observed between reactivity with the reacting antibody of IgG class and the degree of complement-activating activity of the active polysaccharides. The reacting IgG class antibody, which was purified from normal human serum by affinity chromatography on bupleuran 2IIc (a pharmacologically active pectic polysaccharide from the roots of Bupleurum falcatum)-immobilized Sepharose, showed cross-reactivity not only with some other pharmacologically active pectic polysaccharides from other medicinal herbs but also with autoantigens such as single-strand DNA, myosin and tublin from mammals.

Angelica↗

PubMed1997

Structure of the capsular polysaccharide of Clostridium perfringens Hobbs 5 as determined by NMR spectroscopy.

The complete primary structure of the capsular polysaccharide of Clostridium perfringens Hobbs 5, an anaerobic bacterium implicated in food poisoning, was determined. The polysaccharide was isolated from C. perfringens Hobbs 5 cells, after deproteination, by ethanol precipitation and by ion-exchange chromatography. The polysaccharide was comprised of glucose, galactose, mannose, N-acetylglucosamine, N-acetylgalactosamine, and glucuronic acid, in equimolar ratios. Sequence and linkage assignments of the glycosyl residues were obtained by NMR spectroscopy, specifically by the combination of two-dimensional homonuclear TOCSY and NOESY experiments and heteronuclear (1H, 13C) multiple-quantum coherence (HMQC, HMQC-COSY, HMQC-TOCSY and HMBC) experiments. Thus, the envelope polysaccharide of C. perfringens Hobbs 5 was found to be a polymer composed of a hexasaccharide repeating unit with the following structure: [formula: see text] This structure is novel among bacterial cell-surface polysaccharides, and it is the first of many serotypically distinct capsular polysaccharides of C. perfringens to be described.

Bacterial Capsules↗

PubMed1997

Structural studies on the Shigella-like Escherichia coli O121 O-specific polysaccharide.

The O-specific polysaccharide isolated from the lipopolysaccharide (LPS) of Escherichia coli O121 by mild acid hydrolysis has been studied using mainly NMR spectroscopy. The polysaccharide was treated with mild base to yield a O-deacetylated polysaccharide which contained D-GlcNAc, D-GalNAcA, D-GalNAcAN (2-acetamido-2-deoxy-D-galacturonamide) and D-Qui4NAcGly (where D-Qui4N is 4-amino-4,6-dideoxy-D-glucose) in equimolar proportions. The presence of the amide was confirmed by recording the 1H NMR spectrum of the O-deacetylated polysaccharide at different pH values. The O-acetyl group was located on O-3 of the GalNAcAN and the structure of the polysaccharide can be written as [sequence: see text] This structure is almost identical to that previously reported for the O-specific polysaccharide of Shigella dysenteriae type 7 LPS, the only difference being that O-acetylation is stoichiometric in the latter.

Carbohydrate Sequence↗

PubMed2001

Randomized trial of the quantitative and functional antibody responses to a 7-valent pneumococcal conjugate vaccine and/or 23-valent polysaccharide vaccine among HIV-infected adults.

In a double-blinded, randomized trial, human immunodeficiency virus (HIV)-infected adults with > or = 200 CD4 cells/microl received placebo (PL), 7-valent conjugate, or 23-valent pneumococcal polysaccharide (PS) vaccine in one of the following two-dose combinations given 8 weeks apart: conjugate-conjugate, conjugate-polysaccharide, placebo-polysaccharide, placebo-placebo. A total of 67 persons completed the study. Neither significant increases in HIV viral load nor severe adverse reactions occurred in any group. After controlling for confounders, when compared with persons receiving placebo-polysaccharide, persons receiving conjugate-conjugate and conjugate-polysaccharide had higher antibody concentrations (serotypes 4, 6B, 9V and serotype 23F, respectively) and opsonophagocytic titers (functional antibody assay, serotypes 9V, 23F and serotypes 4, 6B, 9V, respectively) after the second dose (P<0.05). The second dose with either conjugate or polysaccharide following the first conjugate dose, however, produced no further increase in immune responses.

Adult↗

PubMed2002

Structure of the O-polysaccharide and classification of Proteus mirabilis strain G1 in Proteus serogroup O3.

The O-chain polysaccharide of the lipopolysaccharide (LPS) of a previously nonclassified strain of Proteus mirabilis termed G1 was studied by sugar analysis and 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, rotating-frame NOE (ROESY), H-detected 1H,13C HMQC, and heteronuclear multiple-bond correlation (HMBC) experiments. The following structure of the polysaccharide was established: [carbohydrate structure: see text] where D-GalA6(L-Lys) stands for N(alpha)-(D-galacturonoyl)-L-lysine. The structure of the O-polysaccharide of P. mirabilis G1 is similar, but not identical, to that of P. mirabilis S1959 and OXK belonging to serogroup O3. Immunochemical studies with P. mirabilis G1 and S1959 anti-(O-polysaccharide) sera revealed close LPS-based serological relatedness of P. mirabilis G1 and S1959, and therefore it was suggested to classify P. mirabilis G1 in serogroup O3 as a subgroup. P. mirabilis G1 and S1959 anti-(O-polysaccharide) sera also cross-reacted with LPS of P. mirabilis strains from two other serogroups containing D-GalA6(L-Lys) in the O-polysaccharide or in the core region.

Animals↗

PubMed2001

Polysaccharide production by a reduced pigmentation mutant of Aureobasidium pullulans NYS-1.

AIMS: To isolate a reduced pigmentation mutant of Aureobasidium pullulans NYS-1 and characterize its cellular pigmentation plus its polysaccharide and biomass production relative to carbon source. METHODS AND RESULTS: Cellular pigmentation, polysaccharide levels and biomass production by the isolated mutant NYSRP-1 were analysed relative to carbon source. Cellular pigmentation of the mutant was lower than its parent strain using either carbon source. The mutant elaborated higher polysaccharide levels on sucrose than on corn syrup. The pullulan content of the polysaccharide synthesized and biomass production by the mutant rose as the carbon source concentration was increased. CONCLUSION: It is feasible to isolate a reduced pigmentation mutant from strain NYS-1 that exhibits elevated polysaccharide production using corn syrup as a carbon source. SIGNIFICANCE AND IMPACT OF THE STUDY: The mutant provides an advantage for commercial pullulan production because of its reduced pigmentation and enhanced polysaccharide synthesis.

Biomass↗

PubMed1991

Antibody response to pneumococcal capsular polysaccharide vaccine in African children.

Antibody titers to six pneumococcal polysaccharides were measured by enzyme-linked immunosorbent assay in Gambian children before and 1 month after vaccination with a 23-valent pneumococcal capsular polysaccharide vaccine at the ages of 2, 4, 6 or 9 months or at 5 to 10 years of age. IgG responses to type 1, 3 and 5 polysaccharides were seen in children in all age groups. Responses to type 19F and 23F polysaccharides were seen only in the oldest children. Few children of any age responded to immunization with type 6A pneumococcal polysaccharide. The immune response of Gambian infants to immunization with six components of a polyvalent pneumococcal polysaccharide vaccine was comparable with that described previously in Finnish infants and would be unlikely to provide substantial protection against invasive pneumococcal disease.

Antibodies, Bacterial↗

PubMed1998

The localization of KpsC, S and T, and KfiA, C and D proteins involved in the biosynthesis of the Escherichia coli K5 capsular polysaccharide: evidence for a membrane-bound complex.

Biosynthesis of the Escherichia coli K5 polysaccharide requires the KfiA, KfiB, KfiC and KfiD proteins. The subsequent transport of the polysaccharide onto the cell surface requires the KpsC, KpsD, KpsE, KpsM, KpsS and KpsT proteins, which are conserved between different group II capsular polysaccharides. The KfiA and KfiC, together with the KpsC, KpsS and KpsT proteins, were purified and polyclonal antisera to each protein generated. These antisera, together with one previously generated (by others) against the purified KfiD protein, were used in Western blot analysis to locate the corresponding proteins within the cell. Analysis of membrane fractions revealed that KfiA (involved in initiation of polysaccharide synthesis), KfiC (K5 glycosyl transferase) and the KfiD protein (UDP-glucose dehydrogenase) were associated with the inner membrane. The KpsC, KpsS, and KpsT proteins involved in polysaccharide transport were associated with the inner membrane and this membrane association occurred in the absence of any other capsule-related proteins. The effect of mutations in individual kps genes on the localization of each protein was determined. Mutations in the kpsC, kpsM, kpsS and kpsT genes resulted in a loss of membrane targeting for KfiA and KfiC, suggesting some form of hetero-oligomeric membrane-bound biosynthetic complex. Osmotic shock caused the release of KfiA, KfiC, KpsC and KpsS from the inner membrane into the periplasm, suggesting that the polysaccharide biosynthetic complex may be associated with sites of adhesion between the inner and outer membrane.

Amino Acid Sequence↗

PubMed1975

Isolation of Polysaccharides Sulfated during Early Embryogenesis in Fucus.

Beginning 10 hours after fertilization, zygotes of Fucus distichus L. Powell incorporate (35)S into polysaccharides as a sulfate ester of fucose. These sulfated polysaccharides are sequestered in only the rhizoid cell of the two-celled embryo and can serve as a marker of cellular differentiation. Zygotes were pulsed at different times after fertilization with Na(2) (35)SO(4) to identify and isolate the fucans localized within the region of cytoplasm destined to become the rhizoid cell. Low molecular weight pools of (35)S were saturated within 60 minutes, with the greatest incorporation into ethanol-soluble and insoluble fractions occurring with 0.1 mm Na(2)SO(4) in the artificial sea water medium. At the time of rhizoid formation, four fucose-containing polysaccharide fractions incorporated (35)S. When each fraction was subjected to diethylaminoethyl chromatography, two components were eluted with KCl that contained over 84% of the fucose and 93% of the (35)S of the particular fraction. Highvoltage paper electrophoresis of each fraction also resulted in the separation of these two major components. Both components from each of the four fractions behaved identically when separated by diethylaminoethyl chromatography and paper electrophoresis. By comparing the incorporation of (35)S into the polysaccharide fractions at 4 and 16 hours after fertilization, the fucan-sulfate components that are localized in the cytoplasm at the time of rhizoid formation were isolated. Although sulfated polysaccharides in brown algae are reported to be very heterogeneous in terms of their sugar composition and complexes with other heteropolymers, we propose that there are two major components that are sulfated during early embryogenesis in Fucus. The location of these two sulfated polysaccharides in different chemical fractions may reflect their subcellular localization (e.g., cytoplasmic vesicles or cell walls), or their association with other heteropolymers.

Journal Article↗