Lymphocytes represent the key antigen-specific leukocyte subpopulation. Despite their importance in mounting an immune response, an unbiased description of proteins expressed by chicken lymphocytes has not been presented. In this study, we therefore intravenously infected chickens with Salmonella Enteritidis, sorted CD4, CD8 and γδ T-lymphocytes from the spleen by flow cytometry and determined the proteome of each population by LC-MS/MS. CD4 T-lymphocyte characteristic proteins included ubiquitin SUMO-like domain and BAR domain containing proteins. CD8 T-lymphocyte specific proteins were characterized by purine ribonucleoside triphosphate binding and were involved in cell differentiation, cell activation and regulation of programmed cell death. γδ T-lymphocyte specific proteins exhibited enrichment of small GTPase of Rab type and GTP binding. Following infection, inducible proteins in CD4 lymphocytes included ribosomal proteins and downregulated proteins localized to the lysosome. CD8 T-lymphocytes induced MCM complex proteins, proteins required for DNA replication and machinery for protein processing in the endoplasmic reticulum. Proteins inducible in γδ T-lymphocytes belonged to immune system response, oxidative phosphorylation and the spliceosome. In this study, we predicted the likely events in lymphocyte response to systemic bacterial infection and identified proteins which can be used as markers specific for each lymphocyte subpopulation.
- MeSH
- CD4-pozitivní T-lymfocyty imunologie metabolismus mikrobiologie MeSH
- CD8-pozitivní T-lymfocyty imunologie metabolismus mikrobiologie MeSH
- intraepiteliální lymfocyty imunologie metabolismus mikrobiologie MeSH
- kur domácí imunologie metabolismus MeSH
- nemoci drůbeže imunologie metabolismus mikrobiologie prevence a kontrola MeSH
- Salmonella enteritidis imunologie metabolismus MeSH
- salmonelové vakcíny imunologie MeSH
- salmonelóza imunologie metabolismus mikrobiologie prevence a kontrola MeSH
- tandemová hmotnostní spektrometrie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In order to design a new Salmonella enterica vaccine, one needs to understand how naive and immune chickens interact differently when exposed to S. enterica. In this study we therefore determined the immune response of vaccinated and non-vaccinated chickens after intravenous infection with Salmonella enterica serovar Enteritidis (S. Enteritidis). Using flow cytometry we showed that 4 days post infection (DPI), counts of CD4 and B-lymphocytes did not change, CD8 and γδ T-lymphocytes decreased and macrophages and heterophils increased in the spleen. When vaccinated and non-vaccinated chickens were compared, only macrophages and heterophils were found in significantly higher counts in the spleens of the non-vaccinated chickens. The non-vaccinated chickens also expressed higher anti-LPS antibodies than the vaccinated chickens. The expression of interleukin (IL)1β, IL6, IL8, IL18, LITAF, IFNγ and iNOS did not exhibit any clear pattern in the cells sorted from the spleens of vaccinated or non-vaccinated chickens. Only IL17 and IL22 showed a differential expression in the CD4 T-lymphocytes of the vaccinated and non-vaccinated chickens at 4 DPI, both being expressed at a higher level in the non-vaccinated chickens. Due to a similar IFNγ expression in the CD4 T-lymphocytes in both the vaccinated and non-vaccinated chickens, and a variable IL17 expression oscillating around IFNγ expression levels, the IL17∶IFNγ ratio in CD4 T-lymphocytes was found to be central for the outcome of the immune response. When IL17 was expressed at higher levels than IFNγ in the non-vaccinated chickens, the Th17 immune response with a higher macrophage and heterophil infiltration in the spleen dominated. However, when the expression of IL17 was lower than that of IFNγ as in the vaccinated chickens, the Th1 response with a higher resistance to S. Enteritidis infection dominated.
- MeSH
- CD4-pozitivní T-lymfocyty cytologie MeSH
- cytokiny metabolismus MeSH
- ELISA metody MeSH
- interferon gama metabolismus MeSH
- interleukin-17 metabolismus MeSH
- kur domácí MeSH
- leukocyty cytologie MeSH
- lipopolysacharidy metabolismus MeSH
- makrofágy metabolismus MeSH
- nemoci drůbeže imunologie prevence a kontrola MeSH
- polymerázová řetězová reakce metody MeSH
- průtoková cytometrie metody MeSH
- Salmonella enteritidis metabolismus MeSH
- salmonelové vakcíny metabolismus MeSH
- salmonelóza imunologie prevence a kontrola MeSH
- slezina cytologie MeSH
- Th1 buňky cytologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Pigs are considered as one of the major sources of zoonotic strains of Salmonella enterica for humans. Out of many S. enterica serovars, S. Typhimurium dominates in pigs, however, in several countries in Central Europe, S. Enteritidis is also quite frequent in pig herds. In this study we therefore compared the colonisation of pigs with S. Typhimurium and S. Enteritidis. We found that 3 weeks after infection S. Enteritidis 147 colonised the intestinal tract in higher quantities but was shed in faeces in lower quantities than S. Typhimurium 17C10. In a second experiment we found out that S. Enteritidis 147 and its SPI-1 and SPI-4 mutants increased proinflammatory cytokine (IL-1β and IL-8) signalling in the ileum 5 days post infection. On the other hand, independent of SPI-1 or SPI-4, S. Enteritidis 147 suppressed expression of IL-18, MCP1, TLR2, CD86, IL-7, IL-10 and IL-15 in the palatine tonsils. The suppression of cytokine signalling may facilitate the initial colonisation of the palatine tonsils by Salmonella. Moreover, immune suppression may also influence pig resistance to opportunistic pathogens and Salmonella infection in pigs thus may become an issue not only in terms of pork contamination but also in terms of affecting the immunological status of pig herds.
- MeSH
- cytokiny imunologie metabolismus MeSH
- krční mandle imunologie mikrobiologie MeSH
- lidé MeSH
- maso MeSH
- nemoci prasat imunologie mikrobiologie MeSH
- prasata MeSH
- Salmonella enteritidis metabolismus fyziologie MeSH
- Salmonella typhimurium metabolismus fyziologie MeSH
- salmonelová infekce u zvířat imunologie mikrobiologie MeSH
- Sus scrofa MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Evropa 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
- MeSH
- Caco-2 buňky enzymologie mikrobiologie sekrece MeSH
- elektrická impedance klasifikace MeSH
- ELISA metody využití MeSH
- enterocyty fyziologie MeSH
- financování vládou MeSH
- interleukin-8 škodlivé účinky MeSH
- Lactobacillus enzymologie metabolismus MeSH
- lidé MeSH
- Salmonella enteritidis enzymologie metabolismus MeSH
- Check Tag
- lidé MeSH