Salmonella Enteritidis is the main serovar of poultry origin in humans, but its complex interaction with certain avian cells is still not fully understood. Previously we identified several genes significantly induced in chicken embryo fibroblasts (CEFs) by the wild-type strain S. Enteritidis 11 (SE 11). In the present study, we raised the question whether virulence-attenuated mutants of this strain would induce altered expression of the newly identified fibroblast genes associated with immune and non-immune functions of CEFs. Gene expression was evaluated by real-time PCR following challenge by the parental strain SE 11 and its virulence attenuated mutants lacking flagellin gene fliD only or fliD and the serovar-specific virulence plasmid pSEV. As a result, deletion mutants induced a lower expression of all immune genes, but an increased expression of the non-immune genes G0S2 and ENO2 relative to the parental strain. Our data indicate the importance of flagella and pSEV in modulation of virulence and host response in this model. We demonstrated, for the first time ever, an increased induction of survival genes G0S2 and ENO2 by virulence-attenuated mutants of S. Enteritidis.
- MeSH
- bakteriální proteiny genetika MeSH
- fibroblasty mikrobiologie MeSH
- flagella genetika MeSH
- interakce hostitele a patogenu MeSH
- kuřecí embryo MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- nemoci drůbeže imunologie mikrobiologie MeSH
- plazmidy genetika MeSH
- Salmonella enteritidis genetika patogenita MeSH
- salmonelová infekce u zvířat imunologie mikrobiologie MeSH
- virulence genetika MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Pathogenic and non-pathogenic related microorganisms differ in secondary metabolite production. Here we show that riboflavin overproduction by a fungal pathogen and its hyperaccumulation in affected host tissue exacerbates a skin infection to necrosis. In white-nose syndrome (WNS) skin lesions caused by Pseudogymnoascus destructans, maximum riboflavin concentrations reached up to 815 μg ml(-1), indicating bioaccumulation and lack of excretion. We found that high riboflavin concentrations are cytotoxic under conditions specific for hibernation, affect bats' primary fibroblasts and induce cell detachment, loss of mitochondrial membrane potential, polymerization of cortical actin, and cell necrosis. Our results explain molecular pathology of WNS, where a skin infection becomes fatal. Hyperaccumulation of vitamin B2 coupled with reduced metabolism and low tissue oxygen saturation during hibernation prevents removal of excess riboflavin in infected bats. Upon reperfusion, oxygen reacts with riboflavin resulting in dramatic pathology after arousal. While multiple molecules enable invasive infection, riboflavin-associated extensive necrosis likely contributes to pathophysiology and altered arousal pattern in infected bats. Bioaccumulation of a vitamin under natural infection represents a novel condition in a complex host-pathogen interplay.
- MeSH
- Ascomycota klasifikace genetika patogenita MeSH
- buněčná adheze MeSH
- Chiroptera mikrobiologie MeSH
- dermatomykózy mikrobiologie MeSH
- elektronová mikroskopie MeSH
- faktory virulence metabolismus MeSH
- fibroblasty cytologie metabolismus mikrobiologie MeSH
- fylogeneze MeSH
- interakce hostitele a patogenu MeSH
- křídla zvířecí cytologie mikrobiologie ultrastruktura MeSH
- kultivované buňky MeSH
- membránový potenciál mitochondrií MeSH
- riboflavin metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Intercellular interactions are able to influence the biological properties of many types of tumors including malignant melanoma. Differentiation pattern of melanoma cells is significantly influenced by the melanoma-associated fibroblasts but the information about interaction of these cells with other important element of melanoma microenvironment, resp. with keratinocytes, is limited. In this, study we tested the effect of fibroblasts isolated from malignant melanoma on phenotype of normal human keratinocytes, especially on their expression of vimentin, a cytoskeletal protein weakly expressed in normal human keratinocytes. The co-culture with normal dermal fibroblasts was used for comparison. The results demonstrated the high expression of vimentin in keratinocytes co-cultured with melanoma-associated fibroblasts compared with those co-cultured with normal dermal fibroblasts. These data suggest participation of melanoma-associated fibroblasts-keratinocyte crosstalk in formation of melanoma niche.
- MeSH
- biomedicínský výzkum metody MeSH
- fenotyp MeSH
- fibroblasty * cytologie mikrobiologie patologie MeSH
- imunohistochemie metody využití MeSH
- interakce genů a prostředí MeSH
- keratinocyty * cytologie enzymologie imunologie MeSH
- kultivační techniky metody využití MeSH
- lidé MeSH
- melanom * genetika imunologie mikrobiologie MeSH
- mezibuněčná komunikace fyziologie genetika imunologie MeSH
- nádorové buňky kultivované cytologie imunologie metabolismus MeSH
- statistika jako téma MeSH
- vimentin imunologie izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria.
- MeSH
- down regulace MeSH
- fibroblasty cytologie metabolismus mikrobiologie MeSH
- kultivované buňky MeSH
- kur domácí genetika metabolismus MeSH
- kuřecí embryo MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- makrofágy metabolismus mikrobiologie MeSH
- nemoci drůbeže metabolismus patologie MeSH
- Salmonella enteritidis fyziologie MeSH
- salmonelová infekce u zvířat metabolismus patofyziologie MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- transkriptom * MeSH
- upregulace MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The modification of implant surface situated in the area of peri-implant sulcus has important role in bacterial and cell adhesion. Six different chemically and physically modified titanium discs were prepared: glazed (Tis-MALP), unglazed (Tis-O), unglazed and alkali-etched (Tis-OA), unglazed and coated with ZrN (Tis-OZ), unglazed, sand blasted, and acid etched (Tis-OPAE), and unglazed, sand blasted, acid, and alkali etched (Tis-OPAAE). Analysis of surface topography was determined using scanning electron microscopy and atomic force microscopy (AFM). Biocompatibility of gingival fibroblasts was characterized by the production of tumor necrosis factor alpha, collagen I, matrix metalloproteinase 2 (MMP-2) after 24 and 72 h and expression of α3 β1 integrin and vinculin using enzyme-linked immunosorbent assay (ELISA) or modified ELISA after 6 and 24 h. Microorganism adhesion (five bacterial strains) and biofilm formation was also evaluated. The adhesion of bacteria and gingival fibroblasts was significantly higher on titanium disc Tis-OPAAE and biofilm formation on the same surface for Streptococcus mutans, Streptococcus gordonii, and Streptococcus intermedius. The gingival fibroblasts on Tis-OPAAE disc had also significantly lower production of MMP-2. The collagen production was significantly lower on all surfaces with roughness higher than 0.2 μm. This study confirmed that the titanium disc with the surface roughness 3.39 μm (Tis-OPAAE) supported the adhesion of bacterial strains as well as gingival fibroblasts.
- MeSH
- bakteriální adheze účinky léků MeSH
- biofilmy účinky léků růst a vývoj MeSH
- biokompatibilní materiály farmakologie MeSH
- buněčná adheze účinky léků MeSH
- fibroblasty cytologie účinky léků mikrobiologie MeSH
- fokální adheze účinky léků metabolismus MeSH
- gingiva cytologie MeSH
- integrin alfa3beta1 metabolismus MeSH
- kolagen typu I biosyntéza MeSH
- lidé MeSH
- matrixová metaloproteinasa 2 biosyntéza MeSH
- mikroskopie atomárních sil MeSH
- mikroskopie elektronová rastrovací MeSH
- spektrometrie rentgenová emisní MeSH
- Streptococcus cytologie účinky léků fyziologie MeSH
- testování materiálů * MeSH
- titan farmakologie MeSH
- TNF-alfa biosyntéza MeSH
- vinkulin metabolismus MeSH
- zubní implantáty mikrobiologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- MeSH
- buněčné dělení účinky léků MeSH
- fibroblasty mikrobiologie účinky léků MeSH
- interferon alfa farmakologie MeSH
- interferon typ I farmakologie MeSH
- ledviny mikrobiologie účinky léků MeSH
- lidé MeSH
- virus vezikulární stomatitidy, kmen Indiana účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
