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PubMedDate not supplied

Effect of Porphyridium sp. polysaccharide on malignant cell transformation by Moloney murine sarcoma virus.

The polysaccharide produced by the red microalga Porphyridium sp. was highly inhibitory for cell transformation of mouse fibroblast cells by an MSV-124 virus stock. This inhibition was most effective if the polysaccharide was added 2 h before or at the time of infection. The finding that the inhibition of cell transformation by MuSV-124 was reversible after removal of the polysaccharide suggested that Porphyridium sp. polysaccharide inhibited a late step after provirus integration into the host genome. Addition of the polysaccharide post-infection significantly reduced the number of transformed cells, but its effect was less marked than that obtained when the polysaccharide was added before or at the time of infection. These findings support the possibility that the polysaccharide may affect early steps in virus replication cycle, such as virus absorption into the host cells, in addition to its effect on a late step after provirus integration.

3T3 Cells↗
PubMed2001

[Studies on polysaccharides in different development stages of Armillaria mellea (Vahl.: Fr.) Quel].

OBJECTIVE: To study the polysaccharides in different development stages of Armillaria mellea. METHOD: Polysaccharides in rhizomorph, fruit-body, mycelia and its fermenting liquor of A. mellea were extracted, isolated and purified. The properties contents, molar ratio and molecular weight of the polysaccharides were determined by IR spectra, HPLC, GPC and gel chromatography. RESULT: The polysaccharides in mycelia and its fermenting liquor contained only glucose, the sugar contents were 9.00% in mycelia and 0.87 g.(100 ml)-1 in fermenting liquor respectively. The polysaccharides in both rhizomorph and fruit-body consisted of glucose and xylose, and the molar ratio was 1:14 in rhizomorph and 1:10 in fruit-body respectively; the polysaccharide contents were 1.12% in rhizomorph and 2.27% in fruit-body. The molecular weight of these polysaccharides was about 10,000-70,000. CONCLUSION: The experimental results supply important scientific data for developing A. mellea as a medicine.

Agaricales↗
PubMed2003

[Experimental study of controlled release microencapsulated Salmonella typhi capsular polysaccharide vaccines immunized mice].

Salmonella Typhi capsular polysaccharide vaccines were encapsulated in the Micro-particles made from polyethylene glycol-poly-DL-lactide (PELA). BALB/c mouse were divided into three groups with 20 mice in each. Mouse were immunized respectively with controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines and Salmonella Typhi capsular polysaccharide vaccines by oral and subcutaneous administration. The mice blood and salvia were collected at the 2nd, 4th and 8th weeks respectively for the titrating of IgG and sIgA antibodies by RIA. At the 8th week, live typhoid bacteria were injected into the immunized mice for the calculation of the rate of immunization protection. The IgG titers of the controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines group were higher than those of the other groups(P < 0.05). The IgA titers of the low groups of controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines (oral and subcutaneous) were higher than those of the group of Salmonella Typhi capsular polysaccharide vaccines (P < 0.05). The immunization protection rates of the three groups were 40%, 100% and 60% respectively. The controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines possess the advantages of releasing slowly in vivo and persisting long time immunogenicity.

Administration, Oral↗
PubMed2004

[The role of Angelica polysaccharides in inducing effector molecule release by peritoneal macrophages].

AIM: To observe the effect of Angelica polysaccharides on effector molecule production by peritoneal macrophages. METHODS: Macrophages were isolated from the peritoneal cavity of BALB/c mice and the primary culture was performed. MTT colorimetry and spectrophotometry were used to examine the effects of Angelica polysaccharides on the releases of effector molecules, such as nitric oxide(NO), tumor necrosis factor-alpha(TNF-alpha), and reactive oxygen species(ROS) as well as inducible nitric oxide synthase (iNOS) and lysozyme(LSZ) activity by peritoneal macrophages. RESULTS: Angelica polysaccharides could promote the releases of NO, TNF-alpha and ROS from macrophages and improved iNOS and LSZ activities in macrophages. However, Angelica polysaccharides had no direct cytotoxicity to tumor cells, but the cultural supernatant of macrophages cocultured with Angelica polysaccharides could kill L929 cells. CONCLUSION: Angelica polysaccharides can promote the releases of NO, TNF-alpha and ROS by macrophages. Angelica polysaccharides may indirectly play the role of anti-tumors through increased TNF-alpha production by macrophages.

Angelica sinensis↗
PubMed2005

[Effect of aloe vera polysaccharide on the release of cytokines and nitric oxide in cultured human keratinocytes].

OBJECTIVE: To investigate the effects of polysaccharide extracted from Aloe Barbadensis on the release of cytokines and nitric oxide (NO) in cultured human keratinocytes. METHODS: The levels of transforming growth factor-alpha (TGF-alpha), TGF-beta1, interleukin-1beta (IL-1beta), IL-6, IL-8, tumor necrosis factor (TNF) and NO in the supernatants of keratinocyte culture in which culture media containing 25, 50, 100, 200, 400 microg/ml, respectively of aloe polysaccharide were assayed. In the control group equal volume of media without the polysaccharide was used. RESULTS: Compared with control group, the levels of TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF in the supernatants of cultured keratinocytes were significantly higher when aloe polysaccharide was added (P<0.05 or P<0.01), and they were positively correlated to the concentration of aloe polysaccharide (P<0.01). However, aloe polysaccharide markedly decreased the level of NO in a dose dependent manner (P<0.01). CONCLUSION: Aloe polysaccharide could promote keratinocytes to secrete TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF, and inhibit the release of NO.

Aloe↗
PubMed2006

Mycelium and polysaccharide production of Agaricus blazei Murrill by submerged fermentation.

BACKGROUND AND PURPOSE: Over the last decade, Agaricus blazei Murrill has been studied and developed as a novel functional food in Japan, Korea, China, and Taiwan. Due to the low yields, the fruiting bodies of A. blazei Murrill are relatively expensive, and a cheap and stable source of A. blazei Murrill mycelium for commercial purposes is highly desirable. Culture media and conditions were investigated with a view to reducing the cost and improving the mycelium and polysaccharide production of A. blazei Murrill by submerged fermentation. METHODS: Thirty six isolates of A. blazei Murrill were isolated from 22 fruiting bodies produced in Taiwan, and 16 of them could be successfully cultivated on mannitol-egg yolk-polymyxin medium. The isolates were identified by species-specific polymerase chain reaction (PCR) and optimized for the culture media and conditions by submerged fermentation for mycelium and polysaccharide production. Some properties of polysaccharide extract were also investigated. RESULTS: All of the PCR products with species-specific primers showed high identity and matched the internal transcribed spacer 1 sequences of A. blazei Murrill. The phylogenic tree of A. blazei Murrill isolates generated from random amplified polymorphic DNAs arranged all samples into 3 groups and 2 independent cases. The optimal culture media of mycelium production in submerged fermentation were 5% malt extract, 0.1% yeast extract, and 0.5% peptone at pH 6.0, while the optimal culture conditions were 200 mL medium in 500 mL Hinton flask, shaking at 90 rpm for 3 days and then shifting to 105 rpm for 5 days at 27 degrees C. Each liter of A. blazei Murrill M72 yielded 10.83 +/- 0.24 g dried mycelia weight and each liter of A. blazei Murrill M152 produced 0.251 +/- 0.004 g crude polysaccharide (3.03 +/- 0.05% of dried mycelia weight). Crude polysaccharide of A. blazei Murrill M162 contained 82.27-99.14% of total sugar and less than 1.63% of protein; it had 4 major molecular weight components (274.1, 32.7, 7.5, and 2.1 kDa, respectively), with the 2.1 kDa portion possibly a beta-(1,3)-glucan. CONCLUSIONS: These results show that selection of media and conditions can be employed in order to improve the mycelium and polysaccharide production of A. blazei Murrill M72 or M152 by submerged fermentation. Mycelia and polysaccharide production of A. blazei Murrill with submerged fermentation is potentially feasible.

Agaricus↗
PubMed1989

The structure and serologic distribution of an extracellular neutral polysaccharide from Pseudomonas aeruginosa immunotype 3.

Previous work has described small molecular weight neutral polysaccharides from isolates of Pseudomonas aeruginosa that appear to be associated with the lipopolysaccharide (LPS) and distributed across serologic barriers defined by antibody to the O side chain. We have isolated and characterized another of these structures obtained from culture supernatants of an immunotype 3 strain of P. aeruginosa. The isolated neutral polysaccharide has a tetrasaccharide repeat unit: (formula; see text) where Rha is rhamnose. The structure was determined by 1H and 13C nuclear magnetic resonance spectroscopy including nuclear Overhauser enhancement experiments, acid hydrolysis, methylation analysis, Smith degradation, and optical rotation determinations. Polyclonal antibodies raised to intact and alkali-treated (0.1 N NaOH, 56 degrees C, 2 h) LPS from the seven Fisher immunotype strains of P. aeruginosa bound well to the neutral polysaccharide. Antibodies affinity purified from these sera using immobilized neutral polysaccharide as well as a neutral polysaccharide-specific monoclonal antibody, E87, reacted with an antigenically similar structure found among many isolates of different LPS serotypes in a colony blot and with LPS from the seven Fisher immunotypes in an immunoblot. In an immunoblot assay, the neutral polysaccharide inhibited binding of the monoclonal antibody, E87, to material present in LPS preparations from a variety of serotypes. This structure may represent another P. aeruginosa neutral polysaccharide variant found associated with the LPS.

Carbohydrate Conformation↗
PubMed1988

Structure of an extracellular cross-reactive polysaccharide from Pseudomonas aeruginosa immunotype 4.

A neutral small molecular mass (approximately 6.5 kDa) polysaccharide comprising a pentasaccharide repeat unit was isolated from culture supernatants of Pseudomonas aeruginosa immunotype 4. The polysaccharide had a pentasaccharide repeating unit as follows (formula; see text) where Rha is rhamnose. The structure was determined using acid hydrolysis, solvolysis with anhydrous hydrogen fluoride, methylation analysis, and 1H and 13C nuclear magnetic resonance spectroscopy including nuclear Overhauser enhancement experiments. The polysaccharide bound antibody raised to the lipopolysaccharide of the seven P. aeruginosa Fisher-Devlin immunotype strains. Inhibition assays demonstrated the presence of a serologically similar polysaccharide in supernatants of these strains. Affinity-purified antibody to the polysaccharide bound to lipopolysaccharide and whole cells of the immunotype strains of P. aeruginosa in a Western immunoblot and colony blot assay, respectively. This polysaccharide seems to contain an antigenic determinant present in the core of the P. aeruginosa lipopolysaccharide or may represent another minor polysaccharide substituent on the lipopolysaccharide in addition to the O side chain.

Carbohydrate Conformation↗
PubMed1987

Evolution of the subclass of IgG antibody to type 3 pneumococcal polysaccharide during childhood.

Human antibodies to bacterial polysaccharides consist primarily of IgG and are largely restricted to the IgG2 subclass in adults. We examined the ontogeny of the IgG subclass response to pneumococcal polysaccharide type 3 to determine if the poor response of infants to immunization with polysaccharide antigens is due to a diminished capacity to form this subclass of antibodies. Sera from 33 patients aged 2 months to 25 years who had previously been shown to respond to polyvalent pneumococcal polysaccharide vaccine by producing IgG antibodies, were assayed for pneumococcal type 3 specific antibodies of the IgG1, IgG2, IgG3, or IgG4 subclass. IgG1 antibodies to pneumococcal polysaccharide type 3 were uniformly low in all age groups. In contrast, IgG2 antibody activity was lowest in children less than the age of 2 years (170 +/- 20 ng/ml), but rose progressively in the age group 2-5 years (210 +/- 40 ng/ml), 5-10 years (330 +/- 30 ng/ml), and over the age of 10 (390 +/- 30 ng/ml) (differences significant at P less than 0.0005 by ANOVA). Thus, even in infants, pneumococcal polysaccharide responses are confined largely to the IgG2 subclass. Our findings are consistent with the hypothesis that purified bacterial capsular polysaccharide antigens preferentially activate IgG2-committed B cell clones at all ages.

Adolescent↗
PubMedDate not supplied

Serum opsonic deficiency produced by Streptococcus pneumoniae and by capsular polysaccharide antigens.

The opsonic requirements for phagocytosis of S. pneumoniae types 6, 7, 18, and 23 were determined in normal and C2 deficient serum, and in normal serum chelated with magnesium ethyleneglycoltetraacetic acid. All four strains were effectively opsonized via the alternative complement pathway, a finding suggesting that the capsular polysaccharides of these strains activated complement via the alternative pathway. Since bacteremic pneumococcal disease is often associated with circulating capsular polysaccharide, it was considered that this cellular component may activate complement in vivo and impair host defenses by producing an opsonic defect for pneumococci. To examine this hypothesis, serum was incubated with suspensions of whole S. pneumoniae types 6, 7, 18, or 23 or with purified capsular polysaccharide from each of these types, and residual complement activity and opsonic capacity were measured. Hemolytic C 3--9 complement activity and opsonic capacity for 3H-thymidine labeled Salmonella typhimurium, a species effectively opsonized via the alternative pathway, were reduced in serum following incubation. Polysaccharide concentrations as low as 1 microgram/ml inhibited serum opsonic capacity for salmonella. Whole pneumococci and pneumococcal capsular polysaccharide also inhibited the opsonic activity of human C2 deficient serum for salmonella, further evidence for activation of complement via the alternative pathway. Pneumococcal capsular polysaccharide markedly inhibited the opsonic capacity of normal serum for the homologous pneumoccal type. Thus, amounts of pneumococcal capsular polysaccharide, similar to those found in the serum of patients with pneumococcal disease, bring about decomplementation of serum via activation of the alternative pathway and inhibit pneumococcal opsonization.

Adult↗
PubMed1986

Surface-bound capsular polysaccharide of type Ia group B Streptococcus mediates C1 binding and activation of the classic complement pathway.

The role of surface-bound type Ia group B Streptococcus (GBS) capsular polysaccharide in antibody-independent binding of C1 and activation of the classic complement pathway was investigated. In a radiolabeled bacterial-polymorphonuclear leukocyte (PMN) association assay, a measure of bacterial opsonization, preincubation of 3H-type Ia GBS with purified F(ab')2 to the organism blocked the association of the bacteria with PMN', and the inhibitory effect was dose dependent. The specificity of F(ab')2 blocking was shown after adsorption of F(ab')2 with type Ia polysaccharide-sensitized erythrocytes. Polysaccharide-adsorbed F(ab')2 had a 70% decrease in ability to block the association of bacteria with PMN. Evidence for the requirement of the capsular polysaccharide in classic complement pathway activation came from a C1 transfer assay with the use of neuraminidase-digested type Ia GBS. Neuraminidase digestion removed 80% of the terminal sialic acid residues from the native polysaccharide. These neuraminidase-digested organisms had a 72% decrease in binding and transfer of purified C1 compared with non-enzyme-treated organisms. Type Ia capsular polysaccharide bound to sheep erythrocytes promoted classic complement pathway-mediated hemolysis of the cells. The role of C1 inhibitor (INH) in modulation of C1 activation by the organisms was investigated. The possibility existed that the C1 INH could be bound by the bacteria, allowing C1 activation to occur in the fluid phase. The inhibitor was purified from human serum, and its activity was measured before and after incubation with type Ia GBS. The organisms had no effect on C1 INH activity. Thus surface-bound capsular polysaccharide of type Ia GBS mediates C1 binding and classic pathway activation, and this does not involve the C1 INH.

Adsorption↗
PubMed1985

Isolation and characterization of a succinylated polysaccharide from the cell wall of Micrococcus agilis.

A polysaccharide consisting of rhamnose, galactose, glucosamine and ester-linked succinic acid was extracted from the isolated cell walls of Micrococcus agilis by the hot water-phenol and 5% trichloroacetic acid (TCA) extraction methods. The hot water-phenol extractable polysaccharide accounted for 30% of the weight of the wall, with 23% by the TCA method. Phosphorus contents were less than 0.01% of the polysaccharide. Succinyl residues released by alkali treatment (0.1 N NaOH, 30 min, 37 degrees C) were identified by gas-liquid chromatography, and accounted for 6.3% and 5.1% of the polysaccharide purified from the hot water-phenol and TCA extracts, respectively. The polysaccharide was not bound when chromatography on Concanavalin A-Sepharose 4B (Con A/Sepharose 4B) columns was performed and it could thus be separated from any residual membrane lipomannan. The purified polysaccharide behaved as a negatively-charged polymer on electrophoresis in 1% agarose (at pH 8.6). A strong cross-reaction, unaffected by removal of the succinyl groups, was observed with type XXIII pneumococcal polysaccharide antiserum indicating the presence of L-rhamnose, linked through non-reducing, lateral end groups.

Antigen-Antibody Complex↗
PubMed1986

IgG2 subclass restriction of antibody to pneumococcal polysaccharides.

Studies in experimental animals suggest that antibody responses to certain polysaccharide antigens may be restricted in IgG subclass distribution. To determine if human antibodies to pneumococcal polysaccharides are similarly restricted we measured the IgG subclass specific response to immunization with purified polyvalent pneumococcal polysaccharide vaccine. For the type 3 pneumococcal antigen, the geometric mean titre of IgG2 antibody was significantly greater than that of IgG1, IgG3 or IgG4, in both pre-immunization and post-immunization sera. A significant rise in mean titre, comparing pre- to post-immunization sera was observed only for IgG2 antibody. Similar predominance of IgG2 antibody was found for pneumococcal polysaccharides types 6, 18, 19 and 23. In contrast, antibody to the protein antigen tetanus toxoid was exclusively of the IgG1 subclass. Patients with IgA/IgG2 deficiency demonstrated a normal IgG response to tetanus, a normal IgM response to pneumococcal polysaccharides, but no IgG antibody to pneumococcal antigens. IgG2 subclass restriction of antibody to pneumococcal polysaccharides suggest that these antigens may elicit an immune response analogous to the murine type 2 T-cell independent immunogens which show IgG subclass restriction and the requirement of a mature B cell subset defined by the Lyb5+ alloantiserum. Our findings support the possibility of subclass-specific inducing or regulating mechanisms for human responses to carbohydrate or polysaccharide antigens.

Adult↗
PubMed1982

Immunoelectron microscopic analysis of polysaccharide and protein surface antigens in wild strains of group B streptococci.

The ultrastructural location of the group polysaccharide, the type-specific polysaccharides and the protein antigens Ibc, R and X of freshly isolated strains of group B streptococci was studied by the direct immunoferritin technique. The results were compared with the findings in prototype strains representing the serological types of Streptococcus agalactiae. In some strains the group antigen was demonstrated over the entire cell surface, but in strains with a large type polysaccharide capsule labelling was confined to the equatorial zone of the cells. If the type polysaccharide was released by extraction, however, the group antigen of these strains was uncovered over the whole surface of the cell. The type polysaccharide capsule was not demonstrated by the conventional electron microscopic technique, but it was visualized after reacting with the homologous type-specific antibody. The extent and density of the polysaccharide capsule were found to differ in strains with the same type antigen. The type polysaccharide did not block demonstration of the simultaneous presence of protein antigens, implying that the two are arranged mosaic-wise inside the capsule.

Antigens, Bacterial↗
PubMed1980

[Isolation of high affinity antibodies to streptococcal group A polysaccharide reacting with thymus and cutaneous epithelial cells].

Anti-streptococcal A polysaccharide (anti-A polysaccharide) antibodies were isolated from sera of rabbits immunized with group A streptococci by means of immunosorbents. The antibodies were studied by indirect immunofluorescence method on sections of skin and thymus tissues. Most preparations of high affinity antibodies reacting with A polysaccharide in the immunodiffusion test also react with skin and thymus epithelial cells. No preparations of low affinity anti-A polysaccharide antibodies reacted with thymus or skin epithelium. It was found that the reaction of antibodies to A-polysaccharide with epithelial cells does not depend on the availability of antibodies cross reacting with group L streptococcal polysaccharide. The reaction with thymus and skin epithelial cells is likely to be bound with high affinity antibodies to the specific determinant of A-polysaccharide.

Animals↗
PubMed1976

The polysaccharides from heterocyst and spore envelopes of a blue-green alga. Methylation analysis and structure of the backbones.

Lindberg's combined gas chromatography-mass spectrometry techniques for the analysis of partially methylated alditol acetate sugar derivatives were used to study the structures of polysaccharides from the envelopes of heterocysts and spores of Anabaena cylindrica. Polysaccharides from both envelopes are highly branched. Glucose, mannose, galactose, and xylose are at terminal positions, whereas glucose and mannose are at internal positions in these polymers. The molar percentages of the 11 partially methylated alditol acetate derivatives observed were approximately the same for both envelopes, suggesting that the envelope polysaccharides may be identical or almost identical. Smith degradation (periodate oxidation followed by reduction with sodium borohydride and mild acid hydrolysis) of the polysaccharides from the two kinds of envelopes removes the side branches without measurable fragmentation of the backbones. Gas chromatographic analysis of partially methylated alditol acetate derivatives of the sugars showed that the backbones of both envelope polysaccharides consist of glucose (Glc) and mannose (Man) linked by 1 leads to 3 glycosidic bonds. Disaccharides, trisaccharides, and tetrasaccharides obtained from the backbone polysaccharides by partial acid hydrolysis were fractionated by column chromatography and separated by high voltage paper electrophoresis. Analysis of these oligosaccharides established that the backbone polysaccharides from both heterocysts and spores consist of repetitions of the structural unit Glc leads to Glc leads to Glc leads to Glc leads to Man, and that all linkages in the backbones are in the beta configuration.

Chromatography, Gas↗
PubMedDate not supplied

[The effect of bacterial polysaccharides on the course of a Mycoplasma infection].

Polysaccharides of Clavibacter michiganense have been studied for their effect on Mycoplasma infection. It is established that content of T-lymphocytes truthfully increased in the groups of animals whose PS 1 and PS 2 were administered both with protective and medical purpose as compared with that in a group of animals with natural course of mycoplasmosis. Under these conditions the effect of polysaccharides on T-helpers were not evidently expressed, while they exerted considerable effect on T-suppressors, normalizing their high index in the animals infected by Mycoplasma hominis. Polysaccharides also exerted a normalizing effect on the content of T-killers, whose level in the animals with natural proceeding of mycoplasmosis was considerably decreased. One can observe a decrease of content of B-lymphocytes (which approached the level of their control in a group of conditionally healthy animals) under the effect of polysaccharides as compared to infected animals. Metabolic processes are activated in immunocompetent cells as affected by polysaccharide. Results obtained when studying the effect of polysaccharides on immunologic factors in the animals with mycoplasma infection as well as histomorphological data confirm the above evidences that polysaccharides of C. michiganense are promising immunomodulators.

Adjuvants, Immunologic↗
PubMed1998

Bactericidal monoclonal antibodies that define unique meningococcal B polysaccharide epitopes that do not cross-react with human polysialic acid.

The poor immunogenicity of the Neisseria meningitidis group B polysaccharide capsule, a homopolymer of alpha(2-->8) sialic acid, has been attributed to immunologic tolerance induced by prenatal exposure to host polysialyated glycoproteins. Substitution of N-propionyl (N-Pr) for N-acetyl groups on the meningococcal B polysaccharide, and conjugation of the resulting polysaccharide to a protein carrier, have been reported to yield a conjugate vaccine that elicits protective Abs with minimal autoantibody activity. To characterize the protective epitopes on the derivatized polysaccharide, we isolated 30 anti-N-Pr meningococcal B polysaccharide mAbs. These Abs were heterogeneous with respect to complement-mediated bactericidal activity, fine antigenic specificity, and autoantibody activity as defined by binding to the neuroblastoma cell line, CHP-134, which expresses long-chain a(2-->8)-linked polysialic acid. Eighteen of the Abs could activate complement-mediated bacteriolysis. Seven of these 18 Abs cross-reacted with N-acetyl meningococcal B polysaccharide by ELISA and had strong autoantibody activity. Thus, N-Pr meningococcal B polysaccharide conjugate vaccine has the potential to elicit autoantibodies. However, 7 of the 18 bactericidal mAbs had no detectable autoantibody activity. These Abs may be useful for the identification of molecular mimetics capable of eliciting protective Abs specific to the bacteria, without the risk of evoking autoimmune disease.

Animals↗