Intra-amniotic infections (IAI) are one of the reasons for preterm birth. High mobility group box 1 (HMGB1) is a nuclear protein with various physiological functions, including tissue healing. Its excessive extracellular release potentiates inflammatory reaction and can revert its action from beneficial to detrimental. We infected the amniotic fluid of a pig on the 80th day of gestation with 1 × 104 colony forming units (CFUs) of E. coli O55 for 10 h, and evaluated the appearance of HMGB1, receptor for glycation endproducts (RAGE), and Toll-like receptor (TLR) 4 in the amniotic membrane and fluid. Sham-infected amniotic fluid served as a control. The expression and release of HMGB1 were evaluated by Real-Time PCR, immunofluorescence, immunohistochemistry, and ELISA. The infection downregulated HMGB1 mRNA expression in the amniotic membrane, changed the distribution of HMGB1 protein in the amniotic membrane, and increased its level in amniotic fluid. All RAGE mRNA, protein expression in the amniotic membrane, and soluble RAGE level in the amniotic fluid were downregulated. TLR4 mRNA and protein expression and soluble TLR4 were all upregulated. HMGB1 is a potential target for therapy to suppress the exaggerated inflammatory response. This controlled expression and release can, in some cases, prevent the preterm birth of vulnerable infants. Studies on suitable animal models can contribute to the development of appropriate therapy.

• MeSH
• amnion imunologie   mikrobiologie   patologie   MeSH
• Escherichia coli růst a vývoj   patogenita   MeSH
• infekce vyvolané Escherichia coli genetika   imunologie   mikrobiologie   veterinární   MeSH
• infekční komplikace v těhotenství genetika   imunologie   mikrobiologie   veterinární   MeSH
• interakce hostitele a patogenu genetika   imunologie   MeSH
• lidé MeSH
• messenger RNA genetika   imunologie   MeSH
• modely nemocí na zvířatech MeSH
• plodová voda imunologie   mikrobiologie   MeSH
• prasata MeSH
• předčasný porod prevence a kontrola   MeSH
• protein HMGB1 genetika   imunologie   MeSH
• receptor pro konečné produkty pokročilé glykace genetika   imunologie   MeSH
• regulace genové exprese MeSH
• signální transdukce MeSH
• těhotenství MeSH
• toll-like receptor 4 genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

High mobility group box 1 (HMGB1) is a DNA-binding nuclear protein that can be actively secreted by immune cells after different immune stimuli or passively released from cells undergoing necrosis. HMGB1 amplifies inflammation, and its hypersecretion contributes to multiple organ dysfunction syndrome and death. We tested possible immunomodulatory effect of commensal Lactobacillus amylovorus (LA), Lactobacillus mucosae (LM) or probiotic Escherichia coli Nissle 1917 (EcN) in infection of gnotobiotic piglets with Salmonella Typhimurium (ST). Transcription of HMGB1 and Toll-like receptors (TLR) 2, 4, and 9 and receptor for advanced glycation end products (RAGE), TLR4-related molecules (MD-2, CD14, and LBP), and adaptor proteins (MyD88 and TRIF) in the ileum and colon were measured by RT-qPCR. Expression of TLR4 and its related molecules were highly upregulated in the ST-infected intestine, which was suppressed by EcN, but not LA nor LM. In contrast, HMGB1 expression was unaffected by ST infection or commensal/probiotic administration. HMGB1 protein levels in the intestine measured by ELISA were increased in ST-infected piglets, but they were decreased by previous colonization with E. coli Nissle 1917 only. We conclude that the stability of HMGB1 mRNA expression in all piglet groups could show its importance for DNA transcription and physiological cell functions. The presence of HMGB1 protein in the intestinal lumen probably indicates cellular damage.

• MeSH
• Escherichia coli imunologie   MeSH
• gnotobiologické modely imunologie   MeSH
• Lactobacillus acidophilus imunologie   MeSH
• prasata * imunologie   mikrobiologie   MeSH
• probiotika * MeSH
• protein HMGB1 imunologie   MeSH
• Salmonella typhimurium imunologie   MeSH
• signální transdukce imunologie   MeSH
• střeva imunologie   mikrobiologie   MeSH
• toll-like receptor 4 imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• alergeny imunologie   MeSH
• aplikace intranazální MeSH
• bronchoalveolární lavážní tekutina cytologie   imunologie   MeSH
• Escherichia coli * MeSH
• myši inbrední BALB C MeSH
• myši knockoutované MeSH
• ovalbumin imunologie   MeSH
• probiotika aplikace a dávkování   MeSH
• protilátky imunologie   MeSH
• respirační alergie imunologie   prevence a kontrola   terapie   MeSH
• toll-like receptor 4 genetika   imunologie   MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH

The protective effect of Enterococcus faecium EFAL41 on chicken's caecum in relation to the TLR (TLR4 and TLR21) activation and production of luminal IgA challenged with Campylobacter jejuni CCM6191 was assessed. The activation of MIF, IFN-β, MD-2 and CD14 was followed-up after bacterial infection. Day-old chicks (40) were divided into four groups (n = 10): control (C), E. faecium AL41 (EFAL41), C. jejuni (CJ) and combined E. faecium AL41+C. jejuni (EFAL41+CJ). Relative mRNA expression of TLR4, TLR21 and CD14 was upregulated in the probiotic strain and infected (combined) group on day 4 and 7 post infection (p.i.). The caecal relative MD-2 mRNA expression was upregulated on day 4 p.i. in the EFAL41+CJ and CJ groups. MIF and IFN-β reached the highest levels in the combined groups on day 7 p.i. The concentration of the sIgA in intestinal flush was upregulated in EFAL41+CJ group on day 4 p.i. The results demonstrated that E. faecium EFAL41 probiotic strain can modulate the TLRs expression and modify the activation of MIF, IFN-β, MD-2 and CD14 molecules in the chickens caecum challenged with C. jejuni CCM 6191. The counts of EFAL41 were sufficient and high, similarly the counts of enterococci in both, caecum and faeces but without reduction of Campylobacter counts.

• MeSH
• antigeny CD14 genetika   imunologie   MeSH
• Campylobacter jejuni růst a vývoj   MeSH
• cékum imunologie   mikrobiologie   MeSH
• Enterococcus faecium růst a vývoj   imunologie   MeSH
• feces mikrobiologie   MeSH
• imunoglobulin A sekreční genetika   MeSH
• interakce hostitele a patogenu MeSH
• interferon beta genetika   imunologie   MeSH
• kampylobakterové infekce dietoterapie   imunologie   mikrobiologie   veterinární   MeSH
• kur domácí MeSH
• lymfocytární antigen 96 genetika   imunologie   MeSH
• nemoci drůbeže dietoterapie   genetika   imunologie   mikrobiologie   MeSH
• novorozená zvířata MeSH
• probiotika farmakologie   MeSH
• protein - isoformy genetika   imunologie   MeSH
• receptory imunologické genetika   imunologie   MeSH
• regulace genové exprese MeSH
• signální transdukce MeSH
• toll-like receptor 4 genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Glycosaminoglycans have anti-inflammatory properties and interact with a variety of soluble and membrane-bound molecules. Little is known about their effects on B cells and humoral immune responses. We show that CS but not dextran or other glycosaminoglycans induces a pronounced proliferation of B cells in vitro compared with TLR4 or TLR9 ligands. With the use of inhibitors and KO mice, we demonstrate that this proliferation is mediated by the tyrosine kinases BTK and Syk but independent of CD44. Antibodies against Ig-α or Ig-β completely block CS-induced B cell proliferation. Injection of CS in mice for 4-5 days expands B cells in the spleen and results in a marked increase of CD138(+) cells in the spleen that is dependent on BTK but independent of CD4(+) T cells. Long-term treatment with CS for 14 days also increases CD138(+) cells in the bone marrow. When mice were immunized with APC or collagen and treated with CS for up to 14 days during primary or after secondary immune responses, antigen-specific humoral immune responses and antigen-specific CD138(+) plasma cells in the bone marrow were reduced significantly. These data show that CD138(+) cells, induced by treatment with CS, migrate into the bone marrow and may displace other antigen-specific plasma cells. Overall, CS is able to interfere markedly with primary and fully established humoral immune responses in mice.

• MeSH
• aktivace lymfocytů účinky léků   genetika   MeSH
• antigeny CD44 genetika   imunologie   MeSH
• buňky kostní dřeně imunologie   MeSH
• chondroitin sulfáty farmakologie   MeSH
• humorální imunita účinky léků   genetika   MeSH
• intracelulární signální peptidy a proteiny genetika   imunologie   MeSH
• myši inbrední BALB C MeSH
• myši knockoutované MeSH
• myši MeSH
• plazmatické buňky imunologie   MeSH
• pohyb buněk účinky léků   imunologie   MeSH
• proliferace buněk účinky léků   MeSH
• syndekan-1 genetika   imunologie   MeSH
• toll-like receptor 4 genetika   imunologie   MeSH
• toll-like receptor 9 genetika   imunologie   MeSH
• tyrosinkinasy genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Deciphering the mechanisms that allow the induction of strong immune responses is crucial to developing efficient vaccines against infectious diseases and cancer. Based on the discovery that the adenylate cyclase from Bordetella pertussis binds to the CD11b/CD18 integrin, we developed a highly efficient detoxified adenylate cyclase-based vector (CyaA) capable of delivering a large variety of Ags to the APC. This vector allows the induction of protective and therapeutic immunity against viral and tumoral challenges as well as against transplanted tumors in the absence of any added adjuvant. Two therapeutic vaccine candidates against human papilloma viruses and melanoma have been developed recently, based on the CyaA vector, and are currently in clinical trials. We took advantage of one of these highly purified vaccines, produced under good manufacturing practice-like conditions, to decipher the mechanisms by which CyaA induces immune responses. In this study, we demonstrate that CyaA binds both human and mouse CD11b(+) dendritic cells (DCs) and induces their maturation, as shown by the upregulation of costimulatory and MHC molecules and the production of proinflammatory cytokines. Importantly, we show that DCs sense CyaA through the TLR4/Toll/IL-1R domain-containing adapter-inducing IFN-β pathway, independent of the presence of LPS. These findings show that CyaA possesses the intrinsic ability to not only target DCs but also to activate them, leading to the induction of strong immune responses. Overall, this study demonstrates that Ag delivery to CD11b(+) DCs in association with TLR4/Toll/IL-1R domain-containing adapter-inducing IFN-β activation is an efficient strategy to promote strong specific CD8(+) T cell responses.

• MeSH
• adaptorové proteiny vezikulární transportní imunologie   MeSH
• adenylátcyklasový toxin imunologie   MeSH
• antigeny CD11b imunologie   MeSH
• antigeny CD80 biosyntéza   MeSH
• antigeny CD86 biosyntéza   MeSH
• Bordetella pertussis imunologie   MeSH
• buněčná diferenciace imunologie   MeSH
• cytotoxické T-lymfocyty imunologie   MeSH
• dendritické buňky cytologie   imunologie   MeSH
• interferon beta imunologie   MeSH
• interleukin-1beta biosyntéza   MeSH
• interleukin-6 biosyntéza   MeSH
• kultivované buňky MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• receptor interferonu alfa-beta genetika   MeSH
• receptory interleukinu-1 imunologie   MeSH
• signální transdukce imunologie   MeSH
• TNF-alfa biosyntéza   MeSH
• toll-like receptor 4 imunologie   MeSH
• tyrosin genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: In vertebrates, it has been repeatedly demonstrated that genes encoding proteins involved in pathogen-recognition by adaptive immunity (e.g. MHC) are subject to intensive diversifying selection. On the other hand, the role and the type of selection processes shaping the evolution of innate-immunity genes are currently far less clear. In this study we analysed the natural variation and the evolutionary processes acting on two genes involved in the innate-immunity recognition of Microbe-Associated Molecular Patterns (MAMPs). RESULTS: We sequenced genes encoding Toll-like receptor 4 (Tlr4) and 7 (Tlr7), two of the key bacterial- and viral-sensing receptors of innate immunity, across 23 species within the subfamily Murinae. Although we have shown that the phylogeny of both Tlr genes is largely congruent with the phylogeny of rodents based on a comparably sized non-immune sequence dataset, we also identified several potentially important discrepancies. The sequence analyses revealed that major parts of both Tlrs are evolving under strong purifying selection, likely due to functional constraints. Yet, also several signatures of positive selection have been found in both genes, with more intense signal in the bacterial-sensing Tlr4 than in the viral-sensing Tlr7. 92% and 100% of sites evolving under positive selection in Tlr4 and Tlr7, respectively, were located in the extracellular domain. Directly in the Ligand-Binding Region (LBR) of TLR4 we identified two rapidly evolving amino acid residues and one site under positive selection, all three likely involved in species-specific recognition of lipopolysaccharide of gram-negative bacteria. In contrast, all putative sites of LBRTLR7 involved in the detection of viral nucleic acids were highly conserved across rodents. Interspecific differences in the predicted 3D-structure of the LBR of both Tlrs were not related to phylogenetic history, while analyses of protein charges clearly discriminated Rattini and Murini clades. CONCLUSIONS: In consequence of the constraints given by the receptor protein function purifying selection has been a dominant force in evolution of Tlrs. Nevertheless, our results show that episodic diversifying parasite-mediated selection has shaped the present species-specific variability in rodent Tlrs. The intensity of diversifying selection was higher in Tlr4 than in Tlr7, presumably due to structural properties of their ligands.

• MeSH
• druhová specificita MeSH
• fylogeneze MeSH
• molekulární evoluce * MeSH
• Murinae klasifikace   genetika   imunologie   MeSH
• přirozená imunita MeSH
• terciární struktura proteinů MeSH
• toll-like receptor 4 chemie   genetika   imunologie   MeSH
• toll-like receptor 7 chemie   genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Celiac disease (CD) is a gluten-responsive, chronic inflammatory enteropathy. IL-1 cytokine family members IL-1β and IL-18 have been associated with the inflammatory conditions in CD patients. However, the mechanisms of IL-1 molecule activation in CD have not yet been elucidated. We show in this study that peripheral blood mononuclear cells (PBMC) and monocytes from celiac patients responded to pepsin digest of wheat gliadin fraction (PDWGF) by a robust secretion of IL-1β and IL-1α and a slightly elevated production of IL-18. The analysis of the upstream mechanisms underlying PDWGF-induced IL-1β production in celiac PBMC show that PDWGF-induced de novo pro-IL-1β synthesis, followed by a caspase-1 dependent processing and the secretion of mature IL-1β. This was promoted by K+ efflux and oxidative stress, and was independent of P2X7 receptor signaling. The PDWGF-induced IL-1β release was dependent on Nod-like receptor family containing pyrin domain 3 (NLRP3) and apoptosis-associated speck like protein (ASC) as shown by stimulation of bone marrow derived dendritic cells (BMDC) from NLRP3(-/-) and ASC(-/-) knockout mice. Moreover, treatment of human PBMC as well as MyD88(-/-) and Toll-interleukin-1 receptor domain-containing adaptor-inducing interferon-β (TRIF)(-/-) BMDC illustrated that prior to the activation of caspase-1, the PDWGF-triggered signal constitutes the activation of the MyD88/TRIF/MAPK/NF-κB pathway. Moreover, our results indicate that the combined action of TLR2 and TLR4 may be required for optimal induction of IL-1β in response to PDWGF. Thus, innate immune pathways, such as TLR2/4/MyD88/TRIF/MAPK/NF-κB and an NLRP3 inflammasome activation are involved in wheat proteins signaling and may play an important role in the pathogenesis of CD.

• MeSH
• adaptorové proteiny vezikulární transportní genetika   imunologie   MeSH
• celiakie MeSH
• dospělí MeSH
• gliadin chemie   imunologie   MeSH
• inflamasomy účinky léků   genetika   imunologie   MeSH
• interleukin-1beta genetika   imunologie   MeSH
• leukocyty mononukleární účinky léků   imunologie   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mitogenem aktivované proteinkinasy genetika   imunologie   MeSH
• myši knockoutované MeSH
• myši MeSH
• pepsin A MeSH
• peptidové fragmenty farmakologie   MeSH
• primární buněčná kultura MeSH
• protein MyD88 genetika   imunologie   MeSH
• regulace genové exprese účinky léků   imunologie   MeSH
• signální transdukce účinky léků   genetika   imunologie   MeSH
• toll-like receptor 2 genetika   imunologie   MeSH
• toll-like receptor 4 genetika   imunologie   MeSH
• transportní proteiny genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• diabetes mellitus 2. typu diagnóza   enzymologie   prevence a kontrola   MeSH
• dyslipidemie enzymologie   komplikace   metabolismus   MeSH
• financování organizované MeSH
• hmotnostní úbytek fyziologie   MeSH
• hypertenze enzymologie   komplikace   prevence a kontrola   MeSH
• inzulinová rezistence * fyziologie   genetika   imunologie   MeSH
• koagulopatie diagnóza   komplikace   metabolismus   MeSH
• lidé MeSH
• metabolický syndrom diagnóza   komplikace   prevence a kontrola   MeSH
• metformin terapeutické užití   MeSH
• mitogenem aktivované proteinkinasy izolace a purifikace   metabolismus   škodlivé účinky   MeSH
• nádory enzymologie   komplikace   metabolismus   MeSH
• obezita * komplikace   prevence a kontrola   terapie   MeSH
• pohybová aktivita fyziologie   MeSH
• statistika jako téma MeSH
• thiazolidindiony terapeutické užití   MeSH
• toll-like receptor 4 imunologie   izolace a purifikace   metabolismus   MeSH
• Check Tag
• lidé MeSH

Innate immune surveillance in the blood is executed mostly by circulating monocytes, which recognise conserved bacterial molecules such as peptidoglycan and lipopolysaccharide. Toll-like receptors (TLR) play a central role in microbe-associated molecular pattern detection. Here, we compared the differences in TLR expression and cytokine production after stimulation of peripheral blood cells with heat-killed Gram-negative and Gram-positive human pathogens Neisseria meningitidis, Escherichia coli, Staphylococcus aureus and Streptococcus pneumoniae. We found that TLR2 expression is up-regulated on monocytes after stimulation with S. aureus, S. pneumoniae, E. coli and N. meningitidis. Moreover, TLR2 up-regulation was positively associated with increasing concentrations of Gram-positive bacteria, whereas higher concentrations of Gram-negative bacteria, especially E. coli, caused a milder TLR2 expression increase compared with low doses. Cytokines were produced in similar dose-dependent profiles regardless of the stimulatory pathogen; however, Gram-negative pathogens induced higher cytokine levels than Gram-positive ones at same concentrations. These results indicate that Gram-positive and Gram-negative bacteria differ in their dose-dependent patterns of induction of TLR2 and TLR4, but not in cytokine expression.

• MeSH
• aktivace transkripce MeSH
• cytokiny biosyntéza   MeSH
• gramnegativní bakterie genetika   imunologie   MeSH
• grampozitivní bakterie genetika   imunologie   MeSH
• lidé MeSH
• monocyty imunologie   MeSH
• přirozená imunita MeSH
• regulace genové exprese MeSH
• signální transdukce imunologie   MeSH
• toll-like receptor 2 biosyntéza   genetika   imunologie   MeSH
• toll-like receptor 4 biosyntéza   genetika   imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH