Structure determination of<i>Streptococcus suis</i>serotype 2 capsular polysaccharide

Calsteren, Marie‐Rose Van; Gagnon, Fleur; Lacouture, Sonia; Fittipaldi, Nahuel; Gottschalk, M.

doi:10.1139/o09-170

Cited by 88 publications

(114 citation statements)

References 46 publications

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“…Due to its high M w , found to be between 410,000 and 480,000 Da by SEC-MALS (28,30), the native polysaccharide of S. suis type 2 must first be depolymerized into smaller fragments. This reduces polydispersity and yields higher ratios of polysaccharide to protein following coupling.…”

Section: Resultsmentioning

confidence: 99%

“…Bacterial cultures were performed as described by Calzas et al (30). CPS extraction and purification, followed by quality control procedures comprising protein determination with the modified Lowry protein assay kit (Pierce, Rockford, IL), nucleic acid quantification using an ND-1000 spectrometer (NanoDrop, Wilmington, DE), and one-dimensional (1D)/2D 1 H nuclear magnetic resonance (NMR) analysis to ensure purity and identity were performed as described by Van Calsteren et al (28).…”

Section: Methodsmentioning

confidence: 99%

“…In 2010, Van Calsteren et al reported the exact structure of the repeating unit for the serotype 2 CPS (28). The CPS repeating unit is composed of a unique arrangement of 1 rhamnose, 1 glucose, 3 galactoses, 1 N-acetylglucosamine, and 1 sialic acid (also named N-acetylneuraminic acid [Neu5Ac]).…”

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confidence: 99%

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Protection against Streptococcus suis Serotype 2 Infection Using a Capsular Polysaccharide Glycoconjugate Vaccine

et al. 2016

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dStreptococcus suis serotype 2 is an encapsulated bacterium and one of the most important bacterial pathogens in the porcine industry. Despite decades of research for an efficient vaccine, none is currently available. Based on the success achieved with other encapsulated pathogens, a glycoconjugate vaccine strategy was selected to elicit opsonizing anti-capsular polysaccharide (anti-CPS) IgG antibodies. In this work, glycoconjugate prototypes were prepared by coupling S. suis type 2 CPS to tetanus toxoid, and the immunological features of the postconjugation preparations were evaluated in vivo. In mice, experiments evaluating three different adjuvants showed that CpG oligodeoxyribonucleotide (ODN) induces very low levels of anti-CPS IgM antibodies, while the emulsifying adjuvants Stimune and TiterMax Gold both induced high levels of IgGs and IgM. Dose-response trials comparing free CPS with the conjugate vaccine showed that free CPS is nonimmunogenic independently of the dose used, while 25 g of the conjugate preparation was optimal in inducing high levels of anti-CPS IgGs postboost. With an opsonophagocytosis assay using murine whole blood, sera from immunized mice showed functional activity. Finally, the conjugate vaccine showed immunogenicity and induced protection in a swine challenge model. When conjugated and administered with emulsifying adjuvants, S. suis type 2 CPS is able to induce potent IgM and isotype-switched IgGs in mice and pigs, yielding functional activity in vitro and protection against a lethal challenge in vivo, all features of a T cell-dependent response. This study represents a proof of concept for the potential of glycoconjugate vaccines in veterinary medicine applications against invasive bacterial infections.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Protection against Streptococcus suis Serotype 2 Infection Using a Capsular Polysaccharide Glycoconjugate Vaccine

et al. 2016

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“…The structures of type 2 and 14 S. suis CPSs are composed of the monosaccharides glucose, galactose, N-acetylglucosamine, and rhamnose (for type 2 only) arranged into a unique repeating unit that also contains a side chain terminated by sialic acid. In fact, these streptococci are the sole Gram-positive bacteria possessing sialic acid in their capsules (8,10,11). However, despite similarities in the compositions of the CPSs of these two bacterial species, each CPS is composed of a unique arrangement of these sugars conferring a distinct antigenicity.…”

Section: S Treptococcus Agalactiae (Also Known As Group B Streptococcusmentioning

confidence: 99%

Group B Streptococcus and Streptococcus suis Capsular Polysaccharides Induce Chemokine Production by Dendritic Cells via Toll-Like Receptor 2- and MyD88-Dependent and -Independent Pathways

et al. 2013

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bStreptococcus agalactiae (also known as group B Streptococcus [GBS]) and Streptococcus suis are encapsulated streptococci causing severe septicemia and meningitis. Bacterial capsular polysaccharides (CPSs) are poorly immunogenic, but anti-CPS antibodies are essential to the host defense against encapsulated bacteria. The mechanisms underlying anti-CPS antibody responses are not fully elucidated, but the biochemistry of CPSs, particularly the presence of sialic acid, may have an immunosuppressive effect. We investigated the ability of highly purified S. suis and GBS native (sialylated) CPSs to activate dendritic cells (DCs), which are crucial actors in the initiation of humoral immunity. The influence of CPS biochemistry was studied using CPSs extracted from different serotypes within these two streptococcal species, as well as desialylated CPSs. No interleukin-1␤ (IL-1␤), IL-6, IL-12p70, tumor necrosis factor alpha (TNF-␣), or IL-10 production was observed in S. suis or GBS CPS-stimulated DCs. Moreover, these CPSs exerted immunosuppressive effects on DC activation, as a diminution of gamma interferon (IFN-␥)-induced B cell-activating factor of the tumor necrosis factor family (BAFF) expression was observed in CPS-pretreated cells. However, S. suis and GBS CPSs induced significant production of CCL3, via partially Toll-like receptor 2 (TLR2)-and myeloid differentiation factor 88 (MyD88)-dependent pathways, and CCL2, via TLR-independent mechanisms. No major influence of CPS biochemistry was observed on the capacity to induce chemokine production by DCs, indicating that DCs respond to these CPSs in a patterned way rather than a structure-dedicated manner.

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“…However, a few strains were weakly hydrophobic, implying that they have completely new cps gene clusters. So far, the CP structures have been determined only for serotypes 2 and 14 (20,21), and cps gene clusters of most of the nonserotypeable isolates have not been analyzed yet. Further analysis, including structure determination of the CPs and cps gene clusters of S. suis strains with different antigenicity, may provide insights into the evolution of S. suis CP as well as the relationship between cps types and serotypes.…”

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Development of a Two-Step Multiplex PCR Assay for Typing of Capsular Polysaccharide Synthesis Gene Clusters of Streptococcus suis

et al. 2014

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iWe developed a practical and easy two-step multiplex PCR assay to aid in serotyping of Streptococcus suis. The assay accurately typed almost all of the serotype reference strains and field isolates of various serotypes and also identified the genotypes of capsular polysaccharide synthesis gene clusters of some serologically nontypeable strains. Streptococcus suis is an important zoonotic pathogen that causes meningitis, septicemia, endocarditis, and other diseases in pigs and humans. S. suis strains have been classified into 35 serotypes (serotypes 1 to 34 and 1/2, which reacts with both serotype 1 and 2 typing sera) (1-4) on the basis of antigenic differences in their capsular polysaccharide (CP) (5). Serotyping of S. suis is one of the most useful methods to understand the epidemiology of a particular outbreak and monitor the prevalence of potentially hazardous strains. However, serotyping with all 35 typing antisera is time-consuming, and preparing the antisera is not easy due to the high cost and labor associated with its production. Additionally, cross-reactions in coagglutination tests with typing antisera and the presence of autoagglutinating strains increase the difficulty of serotyping in some cases. Therefore, the development of more practical and easier serotyping methods is desired.CP synthesis (cps) genes are clustered on a single locus of the chromosome in S. suis (6, 7). We recently sequenced and analyzed the cps gene clusters of all 35 serotype reference strains (8) and reported that 31 (serotypes 3 to 13 and 15 to 34) possessed serotype-specific genes, while the cps gene clusters of serotypes 1 and 14 and serotypes 2 and 1/2 were almost identical in each pair (8). Liu et al. (9) recently developed multiplex PCR assays to target serotype-specific cps genes for the molecular serotyping of S. suis. In their method, the reference strains of 33 serotypes (serotypes 1 to 31, 33, and 1/2) were sorted into their respective serotypes by three multiplex PCR assays, although serotypes 1 and 1/2 were not distinguished from serotypes 14 and 2, respectively (9). In addition, 84 isolates from a patient and clinically healthy pigs, which covered 20 serotypes, were correctly assigned serotypes predicted by coagglutination tests, except for those of a new serotype (serotype 21/29) (9). However, these methods have not yet been validated using field isolates from diseased pigs as well as nontypeable ones by the coagglutination test. Moreover, serotypes 32 and 34 were not included as targets for typing, because the reference strains of these serotypes have been reclassified as Streptococcus orisratti (10). In addition to the serotype 32 and 34 reference strains, those of serotypes 20, 22, 26, and 33 were also recently suggested to be removed from the taxon of S. suis (11). However, whether all isolates of these serotypes actually belong to a species that is different from S. suis remains unclear. From a diagnostic point of view, the isolates of these serotypes recovered from diseased pigs are still identified as S....

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Structure determination ofStreptococcus suisserotype 2 capsular polysaccharide

Cited by 88 publications

References 46 publications

Protection against Streptococcus suis Serotype 2 Infection Using a Capsular Polysaccharide Glycoconjugate Vaccine

Protection against Streptococcus suis Serotype 2 Infection Using a Capsular Polysaccharide Glycoconjugate Vaccine

Group B Streptococcus and Streptococcus suis Capsular Polysaccharides Induce Chemokine Production by Dendritic Cells via Toll-Like Receptor 2- and MyD88-Dependent and -Independent Pathways

Development of a Two-Step Multiplex PCR Assay for Typing of Capsular Polysaccharide Synthesis Gene Clusters of Streptococcus suis

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