Role Of Capsular Polysaccharide Research Articles

Lipopolysaccharides and outer membrane proteins as main structures involved in complement evasion strategies of non-typhoidal Salmonella strains

Non-typhoidal Salmonella (NTS) infections pose a serious public health problem. In addition to the typical course of salmonellosis, an infection with Salmonella bacteria can often lead to parenteral infections and sepsis, which are particularly dangerous for children, the elderly and immunocompromised. Bacterial resistance to serum is a key virulence factor for the development of systemic infections. Salmonella, as an enterobacterial pathogen, has developed several mechanisms to escape and block the antibacterial effects of the complement system. In this review, we discuss the relevance of outer membrane polysaccharides to the complement evasion mechanisms of NTS strains. These include the influence of the overall length and density of the lipopolysaccharide molecules, modifications of the O-antigen lipopolysaccharide composition and the role of capsular polysaccharides in opsonization and protection of the outer membrane from the lytic action of complement. Additionally, we discuss specific outer membrane protein complement evasion mechanisms, such as recruitment of complement regulatory proteins, blocking assembly of late complement components to form the membrane attack complex and the proteolytic cleavage of complement proteins.

Cryptococcus gattii polysaccharide capsule: An insight on fungal-host interactions and vaccine studies.

Cryptococcus neoformans and C. gattii complexes are the main causative agents of cryptococcosis, a neglected disease with high lethality. The capsule, composed predominantly of the capsular polysaccharide (CP) GXM, is the main virulence factor of this pathogen. The role of CP is well described for C. neoformans and; however, there is a scarcity of studies focused on C. gattii, especially in the context of the fungal-host interaction. Understanding how the immune system recognizes C. gattii can generate meaningful information for diagnosing, preventing, and treating cryptococcosis. In the current issue of the European Journal of Immunology [Eur. J. Immunol. 2021. 51: 2281-2295], Ueno etal. demonstrate that CP inhibits C. gattii recognition by CD11b. In this commentary, we highlight the importance of deeply understanding the role of C. gattii CP during infection and how this knowledge would influence the strategies to develop new vaccines against cryptococcosis.

Immunomodulatory Role of Capsular Polysaccharides Constituents of Cryptococcus neoformans.

Cryptococcosis is a systemic fungal infection caused by Cryptococcus neoformans. In immunocompetent patients, cryptococcal infection is often confined to the lungs. In immunocompromised individuals, C. neoformans may cause life-threatening illness, either from novel exposure or through reactivation of a previously acquired latent infection. For example, cryptococcal meningitis is a severe clinical disease that can manifest in people that are immunocompromised due to AIDS. The major constituents of the Cryptococcus polysaccharide capsule, glucuronoxylomannan (GXM), and galactoxylomannan (GalXM), also known as glucuronoxylomanogalactan (GXMGal), are considered the primary virulence factors of Cryptococcus. Despite the predominance of GXM in the polysaccharide capsule, GalXM has more robust immunomodulatory effects on host cellular immunity. This review summarizes current knowledge regarding host-Crytococcus neoformans interactions and the role of capsular polysaccharides in host immunomodulation. Future studies will likely facilitate a better understanding of the mechanisms involved in antigenic recognition and host immune response to C. neoformans and lead to the development of new therapeutic pathways for cryptococcal infection.

Study of Colloidal Polysaccharides Produced by Iron Oxidizing Bacteria Leptospirillum ferriphilum CC

Iron and sulfur oxidizing bacteria produce capsular and colloidal extracellular polymeric substances. The properties and functional role of capsular polysaccharides are well studied. However, colloidal polysaccharides produced by iron oxidizing bacteria have not been sufficiently explored. In this paper, the physical and chemical properties of colloidal polysaccharides produced by the iron oxidizing bacterial isolate Leptospirillum ferriphilum CC have been studied. Colloidal polysaccharides were extracted and further investigated by optical polarized microscopy and analytical programs (LobVIEW15 and NOVA) that allowed determining the size, changes in shape, perimeter, degree of hydration, and crystallization of polysaccharides. Computer modeling of the experimental data has revealed that polysaccharides concentration does not contribute to the size of colloids but influence the number of particles.

Implication of TLR- but not of NOD2-signaling pathways in dendritic cell activation by group B Streptococcus serotypes III and V.

Group B Streptococcus (GBS) is an important agent of life-threatening invasive infection. It has been previously shown that encapsulated type III GBS is easily internalized by dendritic cells (DCs), and that this internalization had an impact on cytokine production. The receptors underlying these processes are poorly characterized. Knowledge on the mechanisms used by type V GBS to activate DCs is minimal. In this work, we investigated the role of Toll-like receptor (TLR)/MyD88 signaling pathway, the particular involvement of TLR2, and that of the intracellular sensing receptor NOD2 in the activation of DCs by types III and V GBS. The role of capsular polysaccharide (CPS, one of the most important GBS virulence factors) in bacterial-DC interactions was evaluated using non-encapsulated mutants. Despite differences in the role of CPS between types III and V GBS in bacterial internalization and intracellular survival, no major differences were observed in their capacity to modulate release of cytokines by DC. For both serotypes, CPS had a minor role in this response. Production of cytokines by DCs was shown to strongly rely on MyD88-dependent signaling pathways, suggesting that DCs recognize GBS and become activated mostly through TLR signaling. Yet, GBS-infected TLR2-/- DCs only showed a partial reduction in the production of IL-6 and CXCL1 compared to control DCs. Surprisingly, CXCL10 release by type III or type V GBS-infected DCs was MyD88-independent. No differences in DC activation were observed between NOD2-/- and control DCs. These results demonstrate the involvement of various receptors and the complexity of the cytokine production pathways activated by GBS upon DC infection.

Role of capsular polysaccharide (CPS) in biofilm formation and regulation of CPS production by quorum‐sensing in Vibrio vulnificus

Extracellular polysaccharides, such as lipopolysaccharide and loosely associated exopolysaccharides, are essential for Vibrio vulnificus to form biofilms. The role of another major component of the V. vulnificus extracellular matrix, capsular polysaccharide (CPS), which contributes to colony opacity, has been characterized in biofilm formation. A CPS-deficient mutant, whose wbpP gene encoding UDP-GlcNAc C4-epimerase was knocked out, formed significantly more biofilm than wild type, due to increased hydrophobicity of the cell surface, adherence to abiotic surfaces and cell aggregation. To elucidate the direct effect of CPS on biofilm structure, extracted CPS and a CPS-degrading enzyme, α-N-acetylgalactosaminidase, were added in biofilm assays, resulting in reduction and increment of biofilm sizes respectively. Therefore, it is suggested that CPS play a critical role in determining biofilm size by restricting continual growth of mature biofilms. Since CPS is required after maturation, CPS biosynthesis should be controlled in a cell density-dependent manner, e.g. by quorum-sensing (QS) regulation. Analysing transcription of the CPS gene cluster revealed that it was activated by SmcR, a QS master regulator, via binding to the upstream region of the cluster. Therefore, CPS was produced when biofilm cell density reached high enough to turn on QS regulation and limited biofilms to appropriate sizes.

Role of Capsular Polysaccharides and Lipooligosaccharides in Campylobacter Surface Properties, Autoagglutination, and Attachment to Abiotic Surfaces

The role of capsular polysaccharides and lipooligosaccharides in cell surface hydrophobicity, surface charge, autoagglutination (AAG), and attachment to abiotic surfaces of three strains of Campylobacter jejuni and one strain of C. coli were investigated. This was achieved by removal of capsular polysaccharides and truncation of lipooligosaccharides core oligosaccharides by inactivation of the kpsE and waaF genes, respectively. The mutants and the wild-type strains were compared after growth under planktonic (broth) and sessile (agar) conditions. Cells grown as planktonic cultures showed a significantly (p<0.05) higher degree of hydrophobicity and AAG activity but differed from their sessile counterparts with respect to surface charge and attachment counts, depending on the strain. These results suggest that prior mode of growth affects the surface properties and attachment of Campylobacter in a strain-dependent manner. There were no significant (p>0.05) differences between the three C. jejuni strains and their ΔkpsE and ΔwaaF mutants with respect to all traits tested. Inactivation of the kpsE gene significantly (p<0.05) reduced the surface charge of the C. coli strain from ∼-10 to ∼-6 mV and increased its AAG activity, while disruption of the waaF gene significantly (p<0.05) increased its surface hydrophobicity by >8° and decreased the numbers of cells attaching to stainless steel and glass by ∼0.5 log/cm². These results suggest that surface polysaccharides may influence the surface properties and attachment to abiotic surfaces of C. coli but not C. jejuni. This suggestion, however, requires further investigation using a larger number of strains of both species.

Role of capsular polysaccharide in Group B Streptococccus interactions with dendritic cells

Group B Streptococcus (GBS) type III is an important agent of life-threatening invasive infections. Albeit the immune system plays a dual role in development and protection against disease, mechanisms leading to an efficient immune response against GBS remain obscure. Mouse bone marrow-derived dendritic cells (DCs) and primary spleen DCs were used to evaluate GBS capacity to modulate the functions of these important antigen-presenting cells. The role of capsular polysaccharide (CPS), one of the most important GBS virulence factors, in bacterial-DC interactions was evaluated by using a non-encapsulated mutant. Phagocytosis assays, confocal and electron microscopy showed that DCs efficiently internalize encapsulated GBS, but the latter possesses strong intracellular survival capacity. GBS devoid of CPS was internalized and killed at higher and faster rates than encapsulated GBS early after infection. Among several cytokines tested, GBS internalization was required for modulation of IL-12, IL-10 and CXCL10 pathways. In contrast, GBS induced DC expression of co-stimulatory molecules in a phagocytosis-independent manner. Finally, the production of pro-inflammatory and Th1 cytokines by GBS-stimulated DCs was differentially modulated by CPS expression, depending on DC origin. Our data suggest multiple mechanisms involved in GBS modulation of DC functions, which were selectively regulated by the presence of CPS.

The role of microbial polysaccharides in host-pathogen interaction

Bacteria are capable of expressing a diverse range of cell surface polysaccharides from capsules and lipopolysaccharides through teichoic acid molecules to lipoarabinomannans. This review will focus on the expression of capsular polysaccharides and their interaction with the host. In particular, it will focus on the role of capsular polysaccharides as immunomodulatory molecules.

Drastic reduction in the virulence of Streptococcus pneumoniae expressing type 2 capsular polysaccharide but lacking choline residues in the cell wall

The role of capsular polysaccharides and several virulence-related proteins in the pathogenic potential of Streptococcus pneumoniae has been studied extensively. Much less information is available about the role of the pneumococcal cell wall in virulence. In this communication we describe an experimental system that has allowed us to test - in a global way - the role of choline, a structural component of the pneumococcal cell wall, in virulence. We constructed double mutants of S. pneumoniae which have lost the auxotrophic requirement for choline and which were also blocked from utilizing choline from the growth medium. Such a double mutant expressing type 2 capsule but completely lacking choline residues from its cell wall grew well both in vitro and also in the blood of infected mice, but showed striking reduction of virulence approaching that of a capsule-free strain in several models of pneumococcal disease including the capacity to attach and invade a human nasopharyngeal cell line; nasal colonization and intraperitoneal and intravenous inoculation in the mouse. The findings allow one to separate the choline requirement of S. pneumoniae into two sharply defined classes: the need for choline in growth and replication which can be effectively bypassed and the need for choline in pneumococcal virulence that appears to be irreplaceable. The double mutant should be a useful experimental tool to dissect the mechanism of choline requirement in various stages of pneumococcal virulence.

Influence of molecular sizes of Cryptococcus neoformans capsular polysaccharide on phagocytosis.

The role of capsular polysaccharides (CPS) of Cryptococcus neoformans in phagocytosis by murine alveolar macrophages was investigated in four strains of C. neoformans serotype A, YC-11, YC-5, YC-27 and YC-13. Phagocytosis rates increased markedly after adding 10% mouse serum, compared to fetal calf serum. The reverse relation between capsular thickness of C. neoformans and phagocytosis by alveolar macrophages was observed except in YC-27, which had thin capsules and high virulence. The phagocytosis rate in mice serum was 17.3% in YC-11 (capsule thickness 2.8-3.5 microns), 39.8% in YC-5 (capsule size 0.8-1.5 microns), 20.3% in YC-27 (capsule size 0.6-1.1 microns), and 62.8% in YC-13 (capsule not detected microscopically). The CPS of YC-11, YC-5, and YC-27 analyzed by gel-filtration using CL-2B showed high molecular fractions near the void volume. However, the CPS of YC-13 showed only low molecular fractions. The widely eluted CPS of YC-11 was separated into 3 fractions and each fraction was added in the phagocytosis assay of YC-13. Phagocytosis was markedly suppressed particularly by the addition of a higher molecular fraction. These results suggest that phagocytosis of C. neoformans by alveolar macrophages is influenced by the molecular sizes of the CPS.

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.

Klebsiella pneumoniae DT-S株によるマウス肺炎発症におけるcapsular polysaccharide及びlipopolysaccharideの役割について

In this study, role of capsular polysaccharide (CPS) and lipopolysaccharide (LPS) of Klebsiella pneumoniae was investigated in experimental mice pneumonia model. Inoculation with K. pneumoniae mucoid strain DT-S into mice lung induced expansive, voluminous lethal pneumonia characterized with thickening of the alveolar septa caused by infiltration of inflammatory cell and packing of bacteria within alveolar spaces. On the other hand, mice lung inoculated with K. pneumoniae DT-X, which was non-mucoid mutant isolated from DT-S during natural passage, showed infiltration of inflammatory cell into alveolar spaces but there was no death of mice during the course of this pneumonia. Inoculation of CPS 100 micrograms of DT-S strain into mice lung induced lesser extent of accumulation of inflammatory cell than that of LPS 4 micrograms of this strain. Stimulation of alveolar and peritoneal macrophage with CPS, even at a concentration of 100 micrograms/ml, induced weaker Interleukin-1 (IL-1) activity than stimulation with LPS 4 micrograms/ml. These results suggest that since CPS of K. pneumoniae DT-S encapsulate bacteria including LPS, CPS may inhibit chemotaxis of inflammatory cell and IL-1 production of macrophage to be induced by LPS during course of pneumonia. It is speculated that existence of CPS have important role in modulating host response to bacterial LPS, and this effect of CPS may be related with difference of pathological findings of lung and lethality between K. pneumoniae DT-S and DT-X.

Studies on nodule formation in locusts following injection of microbial products

The ability of various bacterial cell wall components to induce nodule formation was tested in the locusts, Locusta migratoria and Schistocerca gregaria. Peptidoglycan, particularly the lysozymesoluble fragment, was found to be responsible for the nodule formation elicited by preparations of Micrococcus luteus cell wall. Once extracted, however, the lysozyme-soluble peptidoglycan fragments did not elicit nodules. Isolated teichoic acid extracted from the wall of Staphylococcus aureus also failed to induce nodules. Its removal from cell wall preparations, however, enhanced their ability to trigger nodule formation. A range of Escherichia coli lipopolysaccharide mutants was also tested but there was no clear role for lipopolysaccharide in the nodule formation process. A commercial lipopolysaccharide preparation tested failed to elicit nodule formation. Finally, the role of capsular polysaccharides was examined using a capsulate parent and a noncapsulate mutant strain of Klebsiella pneumoniae. The presence of capsule on the parent strain and capsular material isolated from this strain gave significant enhancement of nodule formation. The results presented are discussed in comparison with the ability of these microbial components to activate prophenoloxidase in vitro. This comparison shows, in the majority of cases, a positive link between prophenoloxidase activation and nodule formation.

Studies on the activation of the prophenoloxidase system of insects by bacterial cell wall components

The ability of bacterial cell walls to activate the prophenoloxidase cascade was tested using Blaberus craniifer, Clitumnus extradentatus, Locusta migratoria and Schistocerca gregaria. Effects of modifying components of the cell wall on the activation of prophenoloxidase in a haemocyte lysate supernatant preparation were examined. Peptidoglycan was found to be an important factor for the activating ability of Gram-positive bacteria. Lysozyme treatment of Micrococcus luteus cell wall showed that the soluble peptidoglycan was the active component. Teichoic acid isolated from Staphylococcus aureus did not activate the prophenoloxidase cascade. However, removal of teichoic acid from the cell wall enhanced activation, probably by exposure of peptidoglycan. Several Escherichia coli K-12 strains, with differing lipopolysaccharide compositions, were also tested for activation of prophenoloxidase. Differences in the ability of the various strains to activate the prophenoloxidase cascade were apparent although no clear conclusions could be made. The role of capsular polysaccharides was investigated too, using two Klebsiella pneumoniae strains, a noncapsulate mutant and its capsulate parent strain. The capsular polysaccharide conferred an increased activating potential. This difference in activation was lost by removal of the capsule from the parent strain. These results are interpreted in terms of the nonself recognition process in insect haemolymph.

Studies on encapsulated lactobacilli—I: Utilization of capsular material by Lactobacillus casei

The role of capsular polysaccharides in the biochemical activities of mucoid (encapsulated) and non-mucoid (non-encapsulated) strains of Lactobacillus casei (L-324M) was investigated. Both the rate and extent of glucose oxidation were greater for non-encapsulated cells although the endogenous oxygen uptake was considerably less than that of encapsulated cells. A positive association was made between endogenous catabolism of capsular polysaccharide as measured by physico-chemical means and the in vitro growth and maintenance of the organism under conditions of nutrient limitation. The taxonomic and serologic implications of the findings are discussed in relation to the oral ecosystem.