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

BACKGROUND: Salmonella enterica serovar Typhimurium is one of the most common enteropathogenic bacteria found in pigs in Europe. In our previous work, we demonstrated the protective effects in suckling piglets when their dams had been vaccinated with an S. Typhimurium-based inactivated vaccine. This study is focused on a procedure leading to serological discrimination between vaccinated and infected pigs. As we supposed, distinct environment during natural infection and in bacterial cultures used for vaccine preparation led to a slightly different spectrum of expressed S. Typhimurium proteins. The examination of porcine antibodies produced after the experimental infection with S. Typhimurium or after vaccination with S. Typhimurium-based inactivated vaccine by affinity chromatography and mass spectrometry revealed differences in antibody response applicable for serological differentiation of infected from vaccinated animals. RESULTS: Antibodies against Salmonella SipB, SipD and SseB proteins were detected at much higher levels in post-infection sera in comparison with control and post-vaccination sera. On the other hand, proteins BamB, OppA and a fragment of FliC interacted with antibodies from post-vaccination sera with a much higher intensity than from control and post-infection sera. In addition, we constructed ELISA assays using post-infection antigen - SipB protein and post-vaccination antigen - FliC-fragment and evaluated them on a panel of individual porcine sera. CONCLUSIONS: The analysis of antibody response of infected and vaccinated pigs by proteomic tools enabled to identify S. Typhimurium antigens useful for distinguishing infected from vaccinated animals. This approach can be utilized in other challenges where DIVA vaccine and a subsequent serological assay are required, especially when genetic modification of a vaccine strain is not desirable.

• MeSH
• antigeny bakteriální metabolismus   MeSH
• ELISA veterinární   MeSH
• inaktivované vakcíny imunologie   MeSH
• nemoci prasat diagnóza   imunologie   MeSH
• prasata MeSH
• proteomika * MeSH
• protilátky bakteriální krev   MeSH
• Salmonella typhimurium genetika   MeSH
• salmonelová infekce u zvířat diagnóza   imunologie   MeSH
• salmonelové vakcíny imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The colonization of poultry with different Salmonella enterica serovars poses an issue throughout the world. In this study we therefore tested the efficacy of a vaccine consisting of attenuated strains of Salmonella enterica serovars Enteritidis, Typhimurium and Infantis against challenge with the same serovars and with S. Agona, Dublin and Hadar. We tested oral and aerosol administration of the vaccine, with or without co-administration of cecal microbiota from adult hens. The protective effect was determined by bacterial counts of the challenge strains up to week 18 of life and by characterizing the immune response using real-time PCR specific for 16 different genes. We have shown that a vaccine consisting of attenuated S. Enteritidis, S. Typhimurium and S. Infantis protected chickens against challenge with the wild type strains of the same serovars and partially protected chickens also against challenge with isolates belonging to serovars Dublin or Hadar. Aerosol vaccination was more effective at inducing systemic immunity whilst oral vaccination stimulated a local immune response in the gut. Co-administration of cecal microbiota increased the protectiveness in the intestinal tract but slightly decreased the systemic immune response. Adjusting the vaccine composition and changing the administration route therefore affects vaccine efficacy.

• MeSH
• atenuované vakcíny terapeutické užití   MeSH
• kombinované vakcíny terapeutické užití   MeSH
• kur domácí imunologie   mikrobiologie   MeSH
• nemoci drůbeže imunologie   prevence a kontrola   MeSH
• Salmonella enteritidis imunologie   MeSH
• Salmonella typhimurium imunologie   MeSH
• Salmonella imunologie   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   prevence a kontrola   MeSH
• salmonelové vakcíny imunologie   terapeutické užití   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Poultry is the most frequent reservoir of non-typhoid Salmonella enterica for humans. Understanding the interactions between chickens and S. enterica is therefore important for vaccine design and subsequent decrease in the incidence of human salmonellosis. In this study we therefore characterized the interactions between chickens and phoP, aroA, SPI1 and SPI2 mutants of S. Enteritidis. First we tested the response of HD11 chicken macrophage-like cell line to S. Enteritidis infection monitoring the transcription of 36 genes related to immune response. All the mutants and the wild type strain induced inflammatory signaling in the HD11 cell line though the response to SPI1 mutant infection was different from the rest of the mutants. When newly hatched chickens were inoculated, the phoP as well as the SPI1 mutant did not induce an expression of any of the tested genes in the cecum. Despite this, such chickens were protected against challenge with wild-type S. Enteritidis. On the other hand, inoculation of chickens with the aroA or SPI2 mutant induced expression of 27 and 18 genes, respectively, including genes encoding immunoglobulins. Challenge of chickens inoculated with these two mutants resulted in repeated induction of 11 and 13 tested genes, respectively, including the genes encoding immunoglobulins. In conclusion, SPI1 and phoP mutants induced protective immunity without inducing an inflammatory response and antibody production. Inoculation of chickens with the SPI2 and aroA mutants also led to protective immunity but was associated with inflammation and antibody production. The differences in interaction between the mutants and chicken host can be used for a more detailed understanding of the chicken immune system.

• MeSH
• atenuované vakcíny imunologie   MeSH
• buněčné linie MeSH
• cékum imunologie   mikrobiologie   MeSH
• kur domácí MeSH
• makrofágy imunologie   MeSH
• mutace MeSH
• nemoci drůbeže imunologie   mikrobiologie   MeSH
• Salmonella enteritidis * genetika   imunologie   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   MeSH
• salmonelové vakcíny imunologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The prevalence of Salmonella enterica serovar Enteritidis is gradually decreasing in poultry flocks in the EU, which may result in the demand for a vaccine that allows for the differentiation of vaccinated flocks from those infected by wild-type S. Enteritidis. In this study, we therefore constructed a (Salmonella Pathogenicity Island 1) SPI1-lon mutant with or without fliC encoding for S. Enteritidis flagellin. The combination of SPI1-lon mutations resulted in attenuated but immunogenic mutant suitable for oral vaccination of poultry. In addition, the vaccination of chickens with the SPI1-lon-fliC mutant enabled the serological differentiation of vaccinated and infected chickens. The absence of fliC therefore did not affect the immunogenicity of the vaccine strain and allowed for serological differentiation of the vaccinated chickens. The SPI1-lon-fliC mutant is therefore a suitable marker vaccine strain for oral vaccination of poultry.

• MeSH
• bakteriální proteiny genetika   imunologie   MeSH
• flagelin genetika   imunologie   MeSH
• kur domácí MeSH
• mutace * MeSH
• nemoci drůbeže prevence a kontrola   MeSH
• proteasa La genetika   imunologie   MeSH
• protilátky bakteriální imunologie   MeSH
• Salmonella enteritidis genetika   růst a vývoj   imunologie   ultrastruktura   MeSH
• salmonelová infekce u zvířat prevence a kontrola   MeSH
• salmonelové vakcíny aplikace a dávkování   imunologie   MeSH
• vakcinace veterinární   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In this study we were interested in the serovar cross-protection potential of Salmonella Pathogenicity Island 1 (SPI1) attenuated vaccine strains of Salmonella enterica serovars Enteritidis and Typhimurium and immune response of vaccinated and naive chickens to Salmonella infection. The immune response was characterized by real time PCR quantifying transcripts of interleukins IL1β, IL17, IL22, interferon gamma (IFNγ), inducible NO synthase (iNOS), immunoglobulins IgM, IgA, IgY and Ig light chain, and six genes of acute phase response including avidin, serum amyloid A, extracellular fatty acid-binding protein (Ex-FABP), immune responsive gene 1, chemokine AH221 and trappin-6. Vaccination with SPI1 mutants of both serovars protected chickens against Salmonella infection, independent of the serovar used for the challenge and the time post infection. However, expressions of all interleukins, iNOS and Ex-FABP showed that protection against homologous serovars was significantly higher than against heterologous serovars after intravenous challenge at 4 days post infection. The vaccination with a mixture of S. Enteritidis and S. Typhimurium SPI1 mutants induced an intermediate protection against challenge with both serovars, i.e. the mixed vaccine provided an additional protective effect when compared with the chickens vaccinated with a vaccine formed by only a single Salmonella serovar.

• MeSH
• atenuované vakcíny genetika   imunologie   MeSH
• chemokiny imunologie   MeSH
• genomové ostrovy MeSH
• kur domácí imunologie   MeSH
• nemoci drůbeže imunologie   prevence a kontrola   MeSH
• protilátky bakteriální krev   imunologie   MeSH
• Salmonella enterica genetika   imunologie   patogenita   MeSH
• Salmonella enteritidis genetika   imunologie   MeSH
• Salmonella typhimurium genetika   imunologie   MeSH
• salmonelová infekce u zvířat imunologie   prevence a kontrola   MeSH
• salmonelové vakcíny genetika   imunologie   MeSH
• vakcinace veterinární   MeSH
• zkřížená ochrana * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem 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 sekrece   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

The present review discusses recent advances, challenges and opportunities for the best use of conjugate vaccines now and in the future. RECENT FINDINGS: Direct protection in young children may be short-lived and programme effectiveness may depend heavily on indirect protection (herd immunity). Pneumococcal carriage serotype replacement has been widely reported following vaccine implementation. Use of pneumococcal conjugate vaccines is being trialled in the elderly. Vaccination in west Africa against Neisseria meningitidis serogroup A (Men A), a new monovalent conjugate vaccine, was commenced in December 2010. New conjugate vaccines against, for example, Salmonella typhi and Streptococcus agalactiae, are being developed and tested in clinical trials. SUMMARY: Conjugate vaccines have been extensively used to immunize children, resulting in significant decreases in childhood morbidity and mortality. Since their introduction, evidence has grown that protection against disease is due to both direct and indirect protection (herd immunity). The optimization of priming and booster dose regimens in existing paediatric vaccination programmes, aiming for maximal and sustained direct and indirect protection using as few doses per child as possible, may broaden conjugate vaccine impact and augment cost-effectiveness in the future. This may be particularly important in strategies for wider global use of conjugate vaccines in children, as well as use in adults and the elderly. Challenges such as pneumococcal serotype replacement make ongoing surveillance of carriage and invasive disease crucial and will have implications for conjugate reformulation and the development of alternative vaccines. New conjugate vaccines for other pathogens currently in clinical trials have the potential to reduce invasive bacterial disease further, particularly in resource-poor settings.

• MeSH
• kolektivní imunita MeSH
• lidé MeSH
• meningokokové vakcíny aplikace a dávkování   imunologie   MeSH
• očkovací schéma MeSH
• pneumokokové vakcíny aplikace a dávkování   imunologie   MeSH
• salmonelové vakcíny aplikace a dávkování   imunologie   MeSH
• sekundární imunizace metody   MeSH
• streptokokové vakcíny aplikace a dávkování   imunologie   MeSH
• vakcinace metody   MeSH
• vakcíny konjugované aplikace a dávkování   imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy 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

If any new live Salmonella vaccine is introduced in the future, it is quite probable that detailed characterisation of its attenuation will be required. In this study we therefore compared 34 isogenic mutants of S. Enteritidis in aroA, aroD, galE, ssrA, sseA, phoP, rpoS, ompR, htrA, clpP, lon, rfaL, rfaG, rfaC, hfq, sodCI, hilA, sipA, avrA, sopB, sopA, sopE, sifA, shdA, fliC, fur, relA, spoT, rel-spoT, misL, rmbA, STM4258, STM4259 and spvBC genes for their resistance to stresses likely to be expected in the host and for their virulence and immunogenicity in Balb/C mice. We found that the cold and bile resistances essentially did not correlate with the resistances to other stress factors. Resistance to acid pH, heat, polymyxin and serum correlated with each other and also with the attenuation. When the residual virulence and immunogenicity were both considered, mutants in htrA, ompR, aroA, aroD and lon performed the best in mice. Furthermore, when a detailed comparison of polymyxin and serum sensitive mutants was performed, the serum sensitive mutants were more immunogenic.

• MeSH
• atenuované vakcíny imunologie   MeSH
• bakteriální geny MeSH
• delece genu MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• polymyxiny imunologie   MeSH
• reprodukovatelnost výsledků MeSH
• Salmonella enteritidis genetika   imunologie   patogenita   MeSH
• salmonelová infekce u zvířat imunologie   prevence a kontrola   MeSH
• salmonelové vakcíny imunologie   MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH