OBJECTIVE: It is an open question whether multifunctional galectin-3 can be a serum marker in inflammatory bowel disease. METHODS: Western blots and commercial ELISA detected and quantitated the lectin immunocytochemistry using double labeling localized it in tissue sections. RESULTS: Serum concentrations were significantly increased in specimen of patients with active and remission-stage ulcerative colitis and Crohn's disease, associated with emerging positivity of CD14(+) cells. CONCLUSION: Enhanced concentration of galectin-3 in serum reflects presence of disease and points to its involvement in the pathogenesis.

• MeSH
• biologické markery MeSH
• Crohnova nemoc krev   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• Escherichia coli metabolismus   MeSH
• fluorescein-5-isothiokyanát MeSH
• fluorescenční barviva MeSH
• galektin 3 krev   MeSH
• idiopatické střevní záněty diagnóza   chemicky indukované   krev   MeSH
• imunohistochemie MeSH
• kolitida chemicky indukované   MeSH
• kolon metabolismus   MeSH
• lektiny metabolismus   MeSH
• lidé MeSH
• lipopolysacharidové receptory analýza   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• síran dextranu MeSH
• ulcerózní kolitida krev   MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

The multifunctional cytokine interleukin-18 (IL-18) is an important mediator in intestinal inflammatory processes. The aim of this study was to evaluate the constitutive expression of IL-18 and its receptors (IL-18Ralpha and IL-18Rbeta) in intestinal epithelial cells (IEC) stimulated by tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). In addition, cellular proliferation and evaluation of brush border enzymes as differentiation markers were studied. Nontransformed rat intestinal epithelial IEC-6 cells were grown on an extracellular matrix (ECM) in medium with or without TNF-alpha, IFN-gamma, or a combination of both. Gene expression of IL-18, its receptors and apoptotic markers was evaluated using real-time PCR. Expression of IL-18Ralpha protein was demonstrated by flow cytometry and Western blot. Enzymatic activities of brush border enzymes and caspase-1 were determined. The constitutive expression of IL-18, IL-18Ralpha and IL-18Rbeta mRNAs and proteins were detected in IEC-6 cells. The biologically active form of IL-18 was released in response to TNF-alpha and IFN-gamma treatment. Exogenous IL-18 had no effect on cellular proliferation, brush border enzyme activities, and gene expression of apoptotic markers. However, the addition of IL-18 stimulated production and release of the chemokine IL-8. These data suggest that IEC-6 cells may be not only a source of IL-18 but also a target for its action.

• MeSH
• apoptóza účinky léků   MeSH
• biologické markery metabolismus   MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčná membrána metabolismus   účinky léků   MeSH
• buněčné linie MeSH
• chemokin CCL2 metabolismus   MeSH
• epitelové buňky MeSH
• financování organizované MeSH
• interferon gama farmakologie   MeSH
• interleukin-18 genetika   sekrece   MeSH
• kaspasa 1 metabolismus   MeSH
• krysa rodu rattus MeSH
• kultivační média MeSH
• messenger RNA genetika   metabolismus   MeSH
• počet buněk MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• receptor interleukinu-18 - alfa-podjednotka genetika   metabolismus   MeSH
• receptor interleukinu-18 - beta-podjednotka genetika   metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• střeva cytologie   MeSH
• TNF-alfa farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

Wheat belongs to six major food allergens inducing IgE-mediated hypersensitivity reaction manifesting as cutaneous, gastrointestinal, and respiratory symptoms. Although cereals are a staple food item in most diets, only a few wheat proteins causing hypersensitivity have been identified. To characterize wheat allergens, salt-soluble wheat extracts were separated by 1-DE and 2-DE and IgE-binding proteins were detected by immunoblotting using sera of patients with allergy to ingested wheat. Proteins, frequently recognized by IgE on 2-DE were analyzed by MALDI-TOF and QTOF and their spectrum was completed by 1-DE and LCQ(DECA) nLC-MS/MS IT technique. Using all three techniques we identified 19 potential wheat allergens such as alpha-amylase inhibitors, beta-amylase, profilin, serpin, beta-D-glucan exohydrolase, and 27K protein. Employing newly developed ELISA, levels of IgE Abs against Sulamit wheat extract and alpha-amylase inhibitors type 1 and 3 were quantified and shown to be significantly elevated in sera of allergic patients compared to those of healthy controls. The level of IgE Abs against alpha-amylase inhibitor type 3 was lower, slightly above the cut-off value in the majority of patients' sera. Our findings contribute to the identification of wheat allergens aimed to increase the specificity of serum IgE and cell activation diagnostic assays.

• MeSH
• 2D gelová elektroforéza MeSH
• alergeny imunologie   MeSH
• alergie na pšenici diagnóza   imunologie   MeSH
• alfa-amylasy antagonisté a inhibitory   MeSH
• dítě MeSH
• dospělí MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• ELISA MeSH
• financování organizované MeSH
• imunoblotting MeSH
• imunoglobulin E krev   MeSH
• inhibitory enzymů imunologie   metabolismus   MeSH
• lidé MeSH
• mladiství MeSH
• peptidové fragmenty analýza   metabolismus   MeSH
• průtoková cytometrie MeSH
• pšenice imunologie   metabolismus   škodlivé účinky   MeSH
• rostlinné proteiny imunologie   metabolismus   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH