The DsbA homolog of Francisella tularensis was previously demonstrated to be required for intracellular replication and animal death. Disruption of the dsbA gene leads to a pleiotropic phenotype that could indirectly affect a number of different cellular pathways. To reveal the broad effects of DsbA, we compared fractions enriched in membrane proteins of the wild-type FSC200 strain with the dsbA deletion strain using a SILAC-based quantitative proteomic analysis. This analysis enabled identification of 63 proteins with significantly altered amounts in the dsbA mutant strain compared to the wild-type strain. These proteins comprise a quite heterogeneous group including hypothetical proteins, proteins associated with membrane structures, and potential secreted proteins. Many of them are known to be associated with F. tularensis virulence. Several proteins were selected for further studies focused on their potential role in tularemia's pathogenesis. Of them, only the gene encoding glyceraldehyde-3-phosphate dehydrogenase, an enzyme of glycolytic pathway, was found to be important for full virulence manifestations both in vivo and in vitro. We next created a viable mutant strain with deleted gapA gene and analyzed its phenotype. The gapA mutant is characterized by reduced virulence in mice, defective replication inside macrophages, and its ability to induce a protective immune response against systemic challenge with parental wild-type strain. We also demonstrate the multiple localization sites of this protein: In addition to within the cytosol, it was found on the cell surface, outside the cells, and in the culture medium. Recombinant GapA was successfully obtained, and it was shown that it binds host extracellular serum proteins like plasminogen, fibrinogen, and fibronectin.

• Klíčová slova
• DsbA, Francisella tularensis, SILAC, glyceraldehyde-3-phosphate dehydrogenase, moonlighting,
• MeSH
• delece genu * MeSH
• faktory virulence analýza   MeSH
• Francisella tularensis enzymologie   imunologie   patogenita   MeSH
• glyceraldehyd-3-fosfátdehydrogenasy nedostatek   metabolismus   MeSH
• krevní proteiny metabolismus   MeSH
• mikrobiální viabilita MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• proteindisulfidisomerasy nedostatek   MeSH
• proteom analýza   MeSH
• salmonelová infekce u zvířat mikrobiologie   patologie   MeSH
• vazba proteinů MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• faktory virulence MeSH
• glyceraldehyd-3-fosfátdehydrogenasy MeSH
• krevní proteiny MeSH
• proteindisulfidisomerasy MeSH
• proteom MeSH

In this study we were interested in the vaccine potential of two attenuated mutants of Salmonella enterica serovar Enteritidis for poultry. The first mutant was attenuated by the removal of the whole Salmonella Pathogenicity Island 1 (SPI1) and the second mutant was devoid of the whole SPI2. These 2 mutants were used for oral vaccination of 2 chicken lines; Lohmann Brown and ISA Brown. Chickens were vaccinated orally on day 1 of life, revaccinated on day 21 and challenged on day 42. The challenge was performed either orally or intravenously. Despite a slightly different response between the two chicken lines, both the mutants gave protection to poultry against S. Enteritidis challenge as documented by findings such as the bacterial counts in tissues, spleen weight, antibody production and cytokine response (namely IL-17 and IL-22). When the 2 mutants were compared, vaccination with the SPI1 mutant proved to be more effective in the protection of poultry against S. Enteritidis challenge than the vaccination with the SPI2 mutant. On the other hand, vaccination with the SPI2 mutant stimulated a slightly higher antibody production and such a mutant might therefore be a better choice if Salmonella is used as a vector for the delivery of heterologous antigens with a desired stimulation of the humoral part of the immune system.

• MeSH
• aplikace orální MeSH
• atenuované vakcíny aplikace a dávkování   genetika   imunologie   MeSH
• bakteriální nálož MeSH
• cytokiny metabolismus   MeSH
• genomové ostrovy * MeSH
• kur domácí MeSH
• leukocyty mononukleární imunologie   MeSH
• nemoci drůbeže imunologie   prevence a kontrola   MeSH
• protilátky bakteriální krev   MeSH
• Salmonella enteritidis imunologie   patogenita   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   patologie   prevence a kontrola   MeSH
• salmonelové vakcíny aplikace a dávkování   genetika   imunologie   MeSH
• sekundární imunizace metody   MeSH
• slezina patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• atenuované vakcíny MeSH
• cytokiny MeSH
• protilátky bakteriální MeSH
• salmonelové vakcíny MeSH

In this study, we were interested in the association of attenuated mutants of Salmonella enterica serovar Enteritidis with subpopulations of porcine white blood cells (WBC). The mutants included those with inactivated aroA, phoP, rfaL, rfaG, rfaC and fliC genes and a mutant with five major pathogenicity islands removed (ΔSPI1-5 mutant). Using flow cytometry, we did not observe any difference in the interactions of the wild-type S. Enteritidis, aroA and phoP mutants with WBC. ΔSPI1-5 and fliC mutants had a minor defect in their association with granulocytes and monocytes, but not with T- or B-lymphocytes. All three rfa mutants associated with granulocytes, monocytes and B-lymphocytes more than the wild-type S. Enteritidis did. Electron microscopy confirmed that the association correlated with the intracellular presence of S. Enteritidis and that the Salmonella-containing vacuole in the WBC infected with the rfa mutants, unlike all other strains, did not develop into a spacious phagosome. Intact lipopolysaccharide, but not the type III secretion system encoded by SPI-1, SPI-2 or the flagellar operon, is important for the initial interaction of S. Enteritidis with porcine leukocytes. This information can be used for the design of live Salmonella vaccines preferentially targeting particular cell types including cancer or tumor cells.

• MeSH
• bakteriální geny genetika   MeSH
• glykosyltransferasy genetika   MeSH
• leukocyty mikrobiologie   ultrastruktura   MeSH
• mutace genetika   MeSH
• nemoci prasat imunologie   mikrobiologie   patologie   MeSH
• prasata MeSH
• Salmonella enteritidis enzymologie   genetika   imunologie   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   patologie   MeSH
• salmonelové vakcíny imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glykosyltransferasy MeSH
• lipooligosaccharide 1,5-heptosyltransferase MeSH Prohlížeč
• salmonelové vakcíny MeSH

Natural variation in the presence or the absence of STM0517-0529 genes allowing allantoin utilisation has been described in field isolates of the multidrug resistant Salmonella enterica serovar Typhimurium belonging to the phage type DT104. Interestingly, S. enterica subspecies enterica serovar Typhimurium DT104 is quite frequent in pigs and cattle, but rarely present in egg-laying hens. Taking into account the different mode of allantoin metabolism in birds and mammals, we were interested in whether the absence of STM0517-0529 genes may disable this clone in poultry colonisation. We have therefore constructed the allB (also designated as STM0523) mutants in S. enterica subspecies enterica serovar Typhimurium and S. enterica subspecies enterica serovar Enteritidis, and with these, we infected mice, newly hatched chickens and adult egg-laying hens to show that the defect in allantoin utilisation does not influence S. enterica virulence for mice or adult hens, but slightly decreases virulence of S. enterica for chickens. The decrease in virulence of the allB mutant was relatively minor as it could be observed only after a mixed infection model, consistent with a lower prevalence, but not a total absence of such clones in poultry flocks.

• MeSH
• alantoin metabolismus   MeSH
• drůbež MeSH
• kyselina močová metabolismus   MeSH
• mnohočetná bakteriální léková rezistence MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nemoci drůbeže mikrobiologie   patologie   MeSH
• Salmonella enteritidis genetika   metabolismus   patogenita   MeSH
• Salmonella typhimurium genetika   metabolismus   patogenita   MeSH
• salmonelová infekce u zvířat mikrobiologie   patologie   MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alantoin MeSH
• kyselina močová MeSH

Chicks (1-d-old, three groups, each containing 50 chicks) were inoculated with 2 x 10(2) and 2 x 10(8) CFU of Salmonella enteritidis; the third group were kept as uninoculated control. Five birds from each group were euthanized at intervals from 6 h to 4 weeks post-inoculation (pi). In the low-dose group S. enteritidis was isolated from 60% cecal samples at 18 h pi, and from 20% of livers at 3 d pi. Individual variation in the frequency of S. enteritidis recovery was observed in this group. The clearance of salmonella from the organs was faster in the low-dose group, and salmonella was not isolated from the liver and cecum at 21 and at 27 d pi, respectively. However, in the high-dose group, S. enteritidis was isolated from all ceca and 80% of liver 6 h pi, and salmonella was detected in the cecum and liver throughout the experiment. Serous typhlitis and unabsorbed yolk sac were the most prevalent lesions in both groups. Granulomatous nodules in the cecum were found occasionally in some cases in both inoculated groups, which can play a role as reservoirs in carrier chicks.

• MeSH
• časové faktory MeSH
• cékum mikrobiologie   patologie   MeSH
• játra mikrobiologie   MeSH
• kur domácí MeSH
• nemoci ptáků mikrobiologie   patologie   MeSH
• Salmonella enteritidis * izolace a purifikace   patogenita   MeSH
• salmonelová infekce u zvířat mikrobiologie   patologie   MeSH
• virulence MeSH
• žloutkový vak patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Reactive NO metabolites play a distinct role in the control of Salmonella enterica serovar Typhimurium (ST; a facultative intracellular pathogen) in susceptible host. A significant increase of nitrite and/or nitrate plasma levels, 3-nitro-tyrosine expression and pathological changes in mesenteric lymph nodes have been observed in gnotobiotic piglets orally infected for 1 d with a virulent strain of ST but not in piglets infected with a rough mutant of ST.

• MeSH
• elektronová mikroskopie MeSH
• fagocytóza MeSH
• gnotobiologické modely MeSH
• imunoenzymatické techniky MeSH
• játra ultrastruktura   MeSH
• lymfatické uzliny metabolismus   ultrastruktura   MeSH
• mezenterium MeSH
• miniaturní prasata MeSH
• oxid dusnatý metabolismus   MeSH
• plíce ultrastruktura   MeSH
• prasata MeSH
• Salmonella typhimurium patogenita   MeSH
• salmonelová infekce u zvířat imunologie   metabolismus   mikrobiologie   patologie   MeSH
• střeva ultrastruktura   MeSH
• tyrosin analogy a deriváty   metabolismus   MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 3-nitrotyrosine MeSH Prohlížeč
• oxid dusnatý MeSH
• tyrosin MeSH

Two stable rough mutants of Salmonella spp. were studied as live peroral vaccines. The SF1591 mutant of S. typhimurium (Ra chemotype) protected germ-free piglets against subsequent infection with virulent smooth S. typhimurium LT2, whereas a deep-rough mutant of S. minnesota mR595 (Re chemotype) did not. We investigated cytokine and leukocyte profiles in the ilea of gnotobiotic piglets colonized for 1 week either with rough mutants alone or with rough mutants followed by S. typhimurium LT2. The ileal mucosae of piglets associated with strain SF1591 alone were not inflamed. Villi contained activated macrophages, and enterocytes expressed transforming growth factor beta (TGF-beta). Subsequent infection of piglets with S. typhimurium LT2 resulted in immigration of alphabeta T cells and immunoglobulin A (IgA) response. In contrast, the ileal mucosae of piglets associated with strain mR595 alone expressed heat shock proteins and inflammatory cytokines but not TGF-beta. Acellular villi contained numerous gammadelta T cells but no alphabeta T cells. After subsequent challenge with the LT2 strain, most piglets died of sepsis. Intestinal mucosae contained IgG but no IgA. These findings suggest the importance of cytokine signals in the regulation of intestinal responses against Salmonella infection.

• MeSH
• cytokiny imunologie   MeSH
• gnotobiologické modely * MeSH
• ileum imunologie   mikrobiologie   patologie   MeSH
• miniaturní prasata MeSH
• prasata MeSH
• Salmonella typhimurium * MeSH
• salmonelová infekce u zvířat imunologie   patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH

The protective effect of dialyzable leukocyte extract (DLE) was investigated in the experimental model of Salmonella infection in calves. DLE was obtained from the lymphatic nodes and spleens of fattening bulls immunized with whole-cell Salmonella vaccine (designated DLEs-im), from the same organs of calves immunized and subsequently infected with Salmonella typhimurium (DLEs-inf), and from non-immunized fattening bulls (DLEn). Three doses of DLEs-inf and DLEs-im applied intravenously at 3-day intervals induced protection in all calves against infection. There were statistically significant differences in the immunological, clinical and microbiological parameters. Three doses of DLEs-inf injected intramuscularly at 3-day intervals provided a protective effect; however, one calf died. The intravenous application of DLEn induced low protection against experimental Salmonella infection and two calves died. The results indicate that the preparation of antigen-specific DLE may be possible via immunization of fattening bulls.

• MeSH
• bakteriální vakcíny MeSH
• inhibice migrace buněk MeSH
• leukocyty imunologie   MeSH
• nemoci skotu imunologie   patologie   prevence a kontrola   MeSH
• Salmonella typhimurium imunologie   MeSH
• salmonelová infekce u zvířat patologie   prevence a kontrola   MeSH
• skot MeSH
• transfer faktor terapeutické užití   MeSH
• vakcinace veterinární   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bakteriální vakcíny MeSH
• transfer faktor MeSH

In one flock, one-eye blindness was observed in the chronical course of disease after infection with the germs of Salmonella arizonae. Changes such as granulomatous iridocyclochorioretinitis with the finding of the bacterial causative agent directly in the pathologically affected tissue were diagnosed on the basis of detailed histological examination of the affected eyeballs. The changes affected almost all parts of the eyeball (iris, ciliary body, lens, retina and chorioid) and were characterized by mononuclear infiltrate with differently intensive admixture of heterophils. The lens was liable to degeneration and fragmentation and giant-cell granulomes of different sizes formed in the posterior chamber.

• MeSH
• krocani * MeSH
• nemoci drůbeže patologie   MeSH
• oči patologie   MeSH
• Salmonella arizonae MeSH
• salmonelová infekce u zvířat patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH

• MeSH
• bursa Fabricii patologie   MeSH
• nekróza MeSH
• nemoci drůbeže patologie   MeSH
• Salmonella typhimurium patogenita   MeSH
• salmonelová infekce u zvířat patologie   MeSH
• slezina patologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH