The LMW surface-layer proteins of Clostridium difficile PCR ribotypes 027 and 001 share common immunogenic properties

Spigaglia, Patrizia; Galeotti, Cesira L.; Barbanti, Fabrizio; Scarselli, María; Broeck, Johan Van; Mastrantonio, Paola

doi:10.1099/jmm.0.029710-0

Cited by 20 publications

(17 citation statements)

References 20 publications

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“…Sequence analysis. SlpA sequence analysis was performed as already described for PR 012, 014/020, 027 and 078 (Spigaglia et al, 2011). We used the same primers designed for the slpA gene of PR 012 to amplify the slpA of PR 045.…”

Section: Methodsmentioning

confidence: 99%

Surface-layer (S-layer) of human and animal Clostridium difficile strains and their behaviour in adherence to epithelial cells and intestinal colonization

Spigaglia

Barketi-Klai

Collignon

et al. 2013

Journal of Medical Microbiology

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Clostridium difficile is a frequent cause of severe, recurrent post-antibiotic diarrhoea and pseudomembranous colitis. The surface layer (S-layer) is the predominant outer surface component of C. difficile which is involved in pathogen-host interactions critical to pathogenesis. In this study, we characterized the S-layer protein A (SlpA) of animal and human strains belonging to different PCRribotypes (PR) and compared the in vitro adherence and in vivo colonization properties of strains showing different SlpA variants. Since each SlpA variant has been recently associated with an S-layer cassette, we were able to deduce the cassette for each of our strains. In this study, an identity of 99-100 % was found among the SlpA of isolates belonging to PR 012, 014/020, 045 and 078. One exception was the SlpA of a poultry isolate, PR 014/020, which showed 99 % identity with that of strain 0160, another PR 014/020 which contains an S-layer cassette 6. Interestingly, this cassette has also been found in a PR 018 strain, an emerging virulent type currently predominant in Italy. Five other SlpA variants (v014/020a-e) were identified in strains PR 014/020. In vitro adherence assays and in vivo colonization experiments were performed on five PR 014/020 strains: human 1064 (v014/020e), human 4684/08 (v014/020b), human IT1106 (v078a), poultry P30 (v014/020d) and poultry PB90 (v014/020b) strains. Adhesion assays indicate that C. difficile strains vary in their capacity to adhere to cells in culture and that adhesion seems to be independent of the SlpA variant. Colonization properties were assessed in vivo using a dixenic mouse model of colonization. The kinetics of faecal shedding and caecal colonization were similar when human 4684/08 (v014/020b) strain was compared with human 1064 (v014/020e) and poultry PB90 (v014/020b) strain. In contrast, poultry P30 (v014/020d) strain outcompeted both human 4684/08 (v014/020b) and IT1106 (v078a) strains and its adherence to caeca at day 7 was significantly higher. The peculiar characteristics of C. difficile P30 seem to advantage it in colonizing the intestinal mice niche, increasing its ability to compete and adapt. The results obtained underline the need of an increased attention to the genetic evolution of C. difficile to prevent and limit the consequences of the emergence of increasingly virulent strains.

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Section: Methodsmentioning

confidence: 99%

Surface-layer (S-layer) of human and animal Clostridium difficile strains and their behaviour in adherence to epithelial cells and intestinal colonization

Spigaglia

Barketi-Klai

Collignon

et al. 2013

Journal of Medical Microbiology

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“…The functions characterized thus far include, e.g., the determination or maintenance of cell shape (Mescher and Strominger 1976; Engelhardt 2007a) and functions as a molecular sieve (Sára and Sleytr 1987; Sára et al 1990), as a binding site for large molecules (Kay et al 1985; Phipps and Kay 1988; Matuschek et al 1994; Egelseer et al 1995, 1996; Peters et al 1995), ions (Schultze-Lam et al 1992; Pollmann et al 2006; Klingl et al 2011) or phages (Howard and Tipper 1973; Ishiguro et al 1984; Fouet 2009) and as a mediator of bacterial adhesion (Doig et al 1992; Toba et al 1995; Noonan and Trust 1997; Hynönen et al 2002; Buck et al 2005; Sakakibara et al 2007; Poppinga et al 2012). In pathogenic bacteria, S-layers may contribute to virulence by several mechanisms, including adhesion, coaggregation (Shimotahira et al 2013), antigenic variation (Thompson 2002; Spigaglia et al 2011), protection from complement or from phagocytosis (Doig et al 1992; Thompson 2002; Shimotahira et al 2013) or modulation of T-cell or cytokine responses (Wang et al 2000; Ausiello et al 2006; Sekot et al 2011; Settem et al 2013). Further, S-layer proteins may protect the bacterial cell from various environmental factors such as mechanical and osmotic stresses (Engelhardt 2007a, b), antimicrobial peptides (de la Fuente-Núñez et al 2012), radiation (Kotiranta et al 1999), changes in environmental pH (Gilmour et al 2000), bacteriophages (Howard and Tipper 1973), bacterial or eukaryotic microbial predators (Koval and Hynes 1991; Tarao et al 2009) or bacteriolytic enzymes (Lortal et al 1992).…”

Section: Introductionmentioning

confidence: 99%

Lactobacillus surface layer proteins: structure, function and applications

Hynönen

Palva

2013

Appl Microbiol Biotechnol

215

196

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Bacterial surface (S) layers are the outermost proteinaceous cell envelope structures found on members of nearly all taxonomic groups of bacteria and Archaea. They are composed of numerous identical subunits forming a symmetric, porous, lattice-like layer that completely covers the cell surface. The subunits are held together and attached to cell wall carbohydrates by non-covalent interactions, and they spontaneously reassemble in vitro by an entropy-driven process. Due to the low amino acid sequence similarity among S-layer proteins in general, verification of the presence of an S-layer on the bacterial cell surface usually requires electron microscopy. In lactobacilli, S-layer proteins have been detected on many but not all species. Lactobacillus S-layer proteins differ from those of other bacteria in their smaller size and high predicted pI. The positive charge in Lactobacillus S-layer proteins is concentrated in the more conserved cell wall binding domain, which can be either N- or C-terminal depending on the species. The more variable domain is responsible for the self-assembly of the monomers to a periodic structure. The biological functions of Lactobacillus S-layer proteins are poorly understood, but in some species S-layer proteins mediate bacterial adherence to host cells or extracellular matrix proteins or have protective or enzymatic functions. Lactobacillus S-layer proteins show potential for use as antigen carriers in live oral vaccine design because of their adhesive and immunomodulatory properties and the general non-pathogenicity of the species.

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“…In addition to secreting the major virulence factors TcdA and TcdB, the vegetative cell expresses other intrinsic immunogenic virulence factors including flagella, surface layer proteins (SLPs) and cell wall proteins (Cwp66, Cwp84 and CwpV) [Ausiello et al 2006;Calabi et al 2002;Drudy et al 2004;Pechine et al 2005b;Tasteyre et al 2001;Janoir et al 2007;Emerson et al 2009]. Surface layer proteins, especially SlpA, can be highly variable across different strains, suggesting that the variable regions of SlpA may have a role in evasion of the host immune response [Bianco et al 2011;Spigaglia et al 2011aSpigaglia et al , 2011b.…”

Section: Intrinsic Antigensmentioning

confidence: 99%

The host immune response to Clostridium difficile infection

Solomon

2013

Therapeutic Advances in Infection

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Clostridium difficile infection (CDI) is the most common infectious cause of healthcare-acquired diarrhoea. Outcomes of C. difficile colonization are varied, from asymptomatic carriage to fulminant colitis and death, due in part to the interplay between the pathogenic virulence factors of the bacterium and the counteractive immune responses of the host.Secreted toxins A and B are the major virulence factors of C. difficile and induce a profound inflammatory response by intoxicating intestinal epithelial cells causing proinflammatory cytokine release. Host cell necrosis, vascular permeability and neutrophil infiltration lead to an elevated white cell count, profuse diarrhoea and in severe cases, dehydration, hypoalbuminaemia and toxic megacolon. Other bacterial virulence factors, including surface layer proteins and flagella proteins, are detected by host cell surface signal molecules that trigger downstream cell-mediated immune pathways.Human studies have identified a role for serum and faecal immunoglobulin levels in protection from disease, but the recent development of a mouse model of CDI has enabled studies into the precise molecular interactions that trigger the immune response during infection. Key effector molecules have been identified that can drive towards a protective anti-inflammatory response or a damaging proinflammatory response.The limitations of current antimicrobial therapies for CDI have led to the development of both active and passive immunotherapies, none of which have, as yet been formally approved for CDI. However, recent advances in our understanding of the molecular basis of host immune protection against CDI may provide an exciting opportunity for novel therapeutic developments in the future.

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The LMW surface-layer proteins of Clostridium difficile PCR ribotypes 027 and 001 share common immunogenic properties

Cited by 20 publications

References 20 publications

Surface-layer (S-layer) of human and animal Clostridium difficile strains and their behaviour in adherence to epithelial cells and intestinal colonization

Surface-layer (S-layer) of human and animal Clostridium difficile strains and their behaviour in adherence to epithelial cells and intestinal colonization

Lactobacillus surface layer proteins: structure, function and applications

The host immune response to Clostridium difficile infection

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