• Publication type
• Meeting Abstract MeSH

We hereby describe the process of design and selection of nonantibody protein binders mimicking cytokine signaling. We chose to mimic signaling of IFN-λ1, type 3 interferon (also known as IL-29) for its novelty and the importance of its biological functions. All four known interferons λ signal through binding to the extracellular domains of IL-28 receptor 1 (IL-28R1) and IL-10 receptor 2 (IL-10R2). Our binders were therefore trained to bind both receptors simultaneously. The bifunctional binder molecules were developed by yeast display, a method of directed evolution. The signaling capacity of the bivalent binders was tested by measuring phosphorylation of the JAK/STAT signaling pathway and production of mRNA of six selected genes naturally induced by IFN- λ1 in human cell lines. The newly developed bivalent binders offer opportunities to study cytokine-related biological functions and modulation of the cell behavior by receptor activation on the cell surfaces alternative to the use of natural IFN-λ.

• MeSH
• Antiviral Agents metabolism   MeSH
• Cytokines metabolism   MeSH
• Interferons * metabolism   MeSH
• Interleukins * metabolism   MeSH
• Humans MeSH
• Signal Transduction MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• Publication type
• Meeting Abstract MeSH

• Publication type
• Meeting Abstract MeSH

Lambda interferons mediate antiviral immunity by inducing interferon-stimulated genes (ISGs) in epithelial tissues. A common variant rs368234815TT/∆G creating functional gene from an IFNL4 pseudogene is associated with the expression of major ISGs in the liver but impaired clearance of hepatitis C. To explain this, we compared Halo-tagged and non-tagged IFNL3 and IFNL4 signaling in liver-derived cell lines. Transfection with non-tagged IFNL3, non-tagged IFNL4 and Halo-tagged IFNL4 led to a similar degree of JAK-STAT activation and ISG induction; however, the response to transfection with Halo-tagged IFNL3 was lower and delayed. Transfection with non-tagged IFNL3 or IFNL4 induced no transcriptome change in the cells lacking either IL10R2 or IFNLR1 receptor subunits. Cytosolic overexpression of signal peptide-lacking IFNL3 or IFNL4 in wild type cells did not interfere with JAK-STAT signaling triggered by interferons in the medium. Finally, expression profile changes induced by transfection with non-tagged IFNL3 and IFNL4 were highly similar. These data do not support the hypothesis about IFNL4-specific non-canonical signaling and point out that functional studies conducted with tagged interferons should be interpreted with caution.

• MeSH
• Cell Line MeSH
• Hep G2 Cells MeSH
• Gene Expression MeSH
• Gene Knockout Techniques MeSH
• Hepatocytes immunology   metabolism   MeSH
• Interferon Regulatory Factors genetics   metabolism   MeSH
• Interferons deficiency   genetics   metabolism   MeSH
• Interleukins deficiency   genetics   metabolism   MeSH
• Humans MeSH
• RNA, Messenger genetics   metabolism   MeSH
• Interleukin-10 Receptor beta Subunit deficiency   genetics   metabolism   MeSH
• Receptors, Interferon deficiency   genetics   metabolism   MeSH
• Recombinant Proteins genetics   metabolism   MeSH
• Signal Transduction MeSH
• Transfection MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Bude provedena validace významu polymorfizmu -764G/C v promotoru genu pro interferon gamma a polymorfismů v genu IL28B pro predikci úspěchu léčby chronické hepatitidy C peginterferonem alfa a ribavirinem na souboru léčených pacientů. U pacientů nově indikovaných k protivirové léčbě chronické hepatitidy C bude stanovena exprese genu pro interferon gamma, interferon lambda 3 a interferon senzitivních genů USP18, ISG 15, IFI27, IFI-6-16 a CXCL9 před zahájením léčby v játrech a v mononukleárech z periferní krve. Vyšetření v mononukleárech bude opakováno v časné fázi léčby.; We shall validate the significance of the polymorphism -764G/C in the interferon gamma gene promoter and polymorphisms in IL28B gene in prediction of treatment response in chronic hepatitis C with peginterferon alpha and ribavirin in a cohort of patients treated in the past. In the group of patients recently starting antiviral treatment of chronic hepatitis C, we shall asses the expression of the interferon gamma gene, interferon lambda 3 and a set of of interferon sensitive genes USP18, ISG15, IFI27, IFI-6-16 and CXCL9 in the liver tissue and peripheral blood mononuclear cells before treatment. The assessment of gene expression will be monitored in peripheral blood mononuclear cells within the early treatment period .

• MeSH
• Antiviral Agents MeSH
• Hepatitis C, Chronic MeSH
• Drug Therapy, Combination MeSH
• Drug Monitoring MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• hepatologie
• terapie
• NML Publication type
• závěrečné zprávy o řešení grantu IGA MZ ČR