Project description:Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus) is a pathogen of infective endocarditis. It was observed previously that this bacterium survives longer in macrophages than other species and the phagocytic uptake by and survival in THP-1 macrophages is strain-dependent.The phagocytosis assay was performed with THP-1 macrophages. S. gallolyticus specific whole genome microarrays were used for transcriptome analysis.Better survival in macrophages was observed for UCN34, BAA-2069 and ATCC43143 than for DSM16831 and LMG17956. S. gallolyticus strains show high resistance to tested bactericidal agents (acid, lysozyme and hydrogen peroxide). S. gallolyticus stimulates significant lower cytokine gene expression and causes less lysis of macrophages compared to the control strain Staphylococcus aureus. S. gallolyticus reacts to oxidative burst with a higher gene expression of NADH oxidase initially at the early phase. Expression of genes involved in D-alanylation of teichoic acid, carbohydrate metabolism and transport systems were upregulated thereafter.S. gallolyticus is very resistant to bactericidal agents normally causing degradation of bacteria in phagolysosomes. Additionally, the D-alanylation of teichoic acid is an important factor for survival.
Project description:Background: Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus) is a pathogen of infective endocarditis. It was observed previously that this bacterium survives longer in macrophages than other species and the phagocytic uptake by and survival in THP-1 macrophages is strain-dependent. Methods: The phagocytosis assay was performed with THP-1 macrophages. S. gallolyticus specific whole genome microarrays were used for transcriptome analysis. Results: Better survival in macrophages was observed for UCN34, BAA-2069 and ATCC43143 than for DSM16831 and LMG17956. S. gallolyticus strains show high resistance to tested bactericidal agents (acid, lysozyme and hydrogen peroxide). S. gallolyticus stimulates significant lower cytokine gene expression and causes less lysis of macrophages compared to the control strain S. aureus. S. gallolyticus reacts to oxidative burst with a higher gene expression of NADH oxidase initially at the early phase. Expression of genes involved in D-alanylation of teichoic acid, carbohydrate metabolism and transport systems were upregulated thereafter. Conclusion: S. gallolyticus is very resistant to bactericidal agents normally causing degradation of bacteria in phagolysosomes. Additionally, the D-alanylation of teichoic acid is an important factor for survival.
Project description:Background: Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus) is a pathogen of infective endocarditis. It was observed previously that this bacterium survives longer in macrophages than other species and the phagocytic uptake by and survival in THP-1 macrophages is strain-dependent. Methods: The phagocytosis assay was performed with THP-1 macrophages. S. gallolyticus specific whole genome microarrays were used for transcriptome analysis. Results: Better survival in macrophages was observed for UCN34, BAA-2069 and ATCC43143 than for DSM16831 and LMG17956. S. gallolyticus strains show high resistance to tested bactericidal agents (acid, lysozyme and hydrogen peroxide). S. gallolyticus stimulates significant lower cytokine gene expression and causes less lysis of macrophages compared to the control strain S. aureus. S. gallolyticus reacts to oxidative burst with a higher gene expression of NADH oxidase initially at the early phase. Expression of genes involved in D-alanylation of teichoic acid, carbohydrate metabolism and transport systems were upregulated thereafter. Conclusion: S. gallolyticus is very resistant to bactericidal agents normally causing degradation of bacteria in phagolysosomes. Additionally, the D-alanylation of teichoic acid is an important factor for survival.
Project description:Background: Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus) is a pathogen of infective endocarditis. It was observed previously that this bacterium survives longer in macrophages than other species and the phagocytic uptake by and survival in THP-1 macrophages is strain-dependent. Methods: The phagocytosis assay was performed with THP-1 macrophages. S. gallolyticus specific whole genome microarrays were used for transcriptome analysis. Results: Better survival in macrophages was observed for UCN34, BAA-2069 and ATCC43143 than for DSM16831 and LMG17956. S. gallolyticus strains show high resistance to tested bactericidal agents (acid, lysozyme and hydrogen peroxide). S. gallolyticus stimulates significant lower cytokine gene expression and causes less lysis of macrophages compared to the control strain S. aureus. S. gallolyticus reacts to oxidative burst with a higher gene expression of NADH oxidase initially at the early phase. Expression of genes involved in D-alanylation of teichoic acid, carbohydrate metabolism and transport systems were upregulated thereafter. Conclusion: S. gallolyticus is very resistant to bactericidal agents normally causing degradation of bacteria in phagolysosomes. Additionally, the D-alanylation of teichoic acid is an important factor for survival.
Project description:Background: Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus) is a pathogen of infective endocarditis. It was observed previously that this bacterium survives longer in macrophages than other species and the phagocytic uptake by and survival in THP-1 macrophages is strain-dependent. Methods: The phagocytosis assay was performed with THP-1 macrophages. S. gallolyticus specific whole genome microarrays were used for transcriptome analysis. Results: Better survival in macrophages was observed for UCN34, BAA-2069 and ATCC43143 than for DSM16831 and LMG17956. S. gallolyticus strains show high resistance to tested bactericidal agents (acid, lysozyme and hydrogen peroxide). S. gallolyticus stimulates significant lower cytokine gene expression and causes less lysis of macrophages compared to the control strain S. aureus. S. gallolyticus reacts to oxidative burst with a higher gene expression of NADH oxidase initially at the early phase. Expression of genes involved in D-alanylation of teichoic acid, carbohydrate metabolism and transport systems were upregulated thereafter. Conclusion: S. gallolyticus is very resistant to bactericidal agents normally causing degradation of bacteria in phagolysosomes. Additionally, the D-alanylation of teichoic acid is an important factor for survival.
Project description:Streptococcus gallolyticus subsp. gallolyticus is a commensal bacterium of the human gastrointestinal tract, and a pathogen causing infective endocarditis and other biofilm-associated infections via exposed collagen. This study focuses on the characterization of the biofilm formation and collagen adhesion of S. gallolyticus subsp. gallolyticus under different conditions. In this study, it has been observed that the isolate UCN 34 is resistant to 20 mg/ml lysozyme in BHI medium, whereas the strain BAA-2069 builds more biofilm in the presence of lysozyme compared to in a control of BHI without lysozyme. A transcriptome analysis with whole genome microarrays of these two isolates in BHI medium with lysozyme compared to control without lysozyme revealed changes in gene expression levels. In the isolate BAA-2069, 67 genes showed increased expression in the presence of lysozyme, while in the isolate UCN 34, 165 genes showed increased expression and 30 genes showed decreased expression through lysozyme treatment. Products of genes which were higher expressed are in involved in transcription and translation, in cell-wall modification, in hydrogen peroxide resistance and in bacterial immunity. Furthermore, the adhesion ability of different strains of S. gallolyticus subsp. gallolyticus to collagen type I and IV was analyzed. Thereby, we compared the adhesion of 46 human isolates with 23 isolates from animals. It was shown that the adhesion ability depends significantly on whether the isolate was isolated from human or animal. For example, high adhesion ability was observed for strain UCN 34 isolated from an infective endocarditis patient, whereas strain DSM 16831 isolated from koala feces adhered only marginally to collagen. Full genome microarray analysis of these two strains revealed strain-dependent gene expression due to adhesion. The expression of 25 genes of a transposon and 15 genes of a phage region in strain DSM 16831 were increased, which corresponds to horizontal gene transfer. Adherence to collagen in strain UCN 34 led to higher expression of 27 genes and lower expression of 31 genes. This was suggestive of a change in nutrient uptake.
Project description:Enterococcus faecium and Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus) were classically clustered into the Lancefield Group D streptococci and despite their taxonomic reclassification still share a similar genetic content and environment. Both species are considered as opportunistic pathogens. E. faecium is often associated with nosocomial bacteraemia, and S. gallolyticus is sporadically found in endocarditis of colorectal cancer patients. In both cases, the source of infection is commonly endogenous with a translocation process that launches through the intestinal barrier. To get new insights into the pathological processes preceding infection development of both organisms, we used an in vitro model with Caco-2 cells to study and compare the adhesion, invasion and translocation inherent abilities of 6 E. faecium and 4 S. gallolyticus well-characterized isolates. Additionally, biofilm formation on polystyrene, collagen I and IV was also explored. Overall results showed that E. faecium translocated more efficiently than S. gallolyticus, inducing a destabilization of the intestinal monolayer. Isolates Efm106, Efm121 and Efm113 (p < .001 compared to Ef222) exhibited the higher translocation ability and were able to adhere 2-3 times higher than S. gallolyticus isolates. Both species preferred the collagen IV coated surfaces to form biofilm but the S. gallolyticus structures were more compact (p = .01). These results may support a relationship between biofilm formation and vegetation establishment in S. gallolyticus endocarditis, whereas the high translocation ability of E. faecium high-risk clones might partially explain the increasing number of bacteraemia.
Project description:Streptococcus gallolyticus subsp. gallolyticus (Sg) has long been known to have a strong association with colorectal cancer (CRC). This knowledge has important clinical implications, and yet little is known about the role of Sg in the development of CRC. Here we demonstrate that Sg promotes human colon cancer cell proliferation in a manner that depends on cell context, bacterial growth phase and direct contact between bacteria and colon cancer cells. In addition, we observed increased level of ?-catenin, c-Myc and PCNA in colon cancer cells following incubation with Sg. Knockdown or inhibition of ?-catenin abolished the effect of Sg. Furthermore, mice administered with Sg had significantly more tumors, higher tumor burden and dysplasia grade, and increased cell proliferation and ?-catenin staining in colonic crypts compared to mice receiving control bacteria. Finally, we showed that Sg is present in the majority of CRC patients and is preferentially associated with tumor compared to normal tissues obtained from CRC patients. These results taken together establish for the first time a tumor-promoting role of Sg that involves specific bacterial and host factors and have important clinical implications.