The notion that alternative peptide substrates can be processed and presented to the MHC class I pathway has opened for new aspects on how the immune system detects infected or damaged cells. Recent works show that antigenic peptides are derived from intron sequences in pre-mRNAs target for the nonsense-mediated degradation pathway. Introns are spliced out co-transcriptionally suggesting that such pioneer translation products (PTPs) are synthesized on the nascent RNAs in the nuclear compartment to ensure that the first peptides to emerge from an mRNA are destined for the class I pathway. This illustrates an independent translation event during mRNA maturation that give rise to specific peptide products with a specific function in the immune system. The characterization of the translation apparatus responsible for PTP synthesis will pave the way for understanding how PTP production is regulated in different tissues under different conditions and will help designing new vaccine strategies.

• MeSH
• buněčné jádro genetika   imunologie   MeSH
• CD8-pozitivní T-lymfocyty cytologie   imunologie   MeSH
• cytosol imunologie   metabolismus   MeSH
• dendritické buňky cytologie   imunologie   metabolismus   MeSH
• fagozomy genetika   imunologie   MeSH
• introny MeSH
• lidé MeSH
• MHC antigeny I. třídy genetika   imunologie   MeSH
• peptidy genetika   imunologie   MeSH
• prekurzory RNA genetika   imunologie   MeSH
• prezentace antigenu genetika   MeSH
• proteasomový endopeptidasový komplex genetika   imunologie   MeSH
• proteosyntéza imunologie   MeSH
• sestřih RNA imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

With the growing availability of genomic sequence information, there is an increasing need for gene function analysis. Antibody-mediated "silencing" represents an intriguing alternative for the precise inhibition of a particular function of biomolecules. Here, we describe a method for selecting recombinant antibodies with a specific purpose in mind, which is to inhibit intrinsic protein-protein interactions in the cytosol of plant cells. Experimental procedures were designed for conveniently evaluating desired properties of recombinant antibodies in consecutive steps. Our selection method was successfully used to develop a recombinant antibody inhibiting the interaction of ARABIDOPSIS HISTIDINE PHOSPHOTRANSFER PROTEIN 3 with such of its upstream interaction partners as the receiver domain of CYTOKININ INDEPENDENT HISTIDINE KINASE 1. The specific down-regulation of the cytokinin signaling pathway in vivo demonstrates the validity of our approach. This selection method can serve as a prototype for developing unique recombinant antibodies able to interfere with virtually any biomolecule in the living cell.

• MeSH
• Arabidopsis genetika   MeSH
• cytosol imunologie   metabolismus   MeSH
• fosfotransferasy biosyntéza   genetika   imunologie   MeSH
• mapy interakcí proteinů genetika   imunologie   MeSH
• proteinkinasy biosyntéza   genetika   imunologie   MeSH
• proteiny huseníčku biosyntéza   genetika   imunologie   MeSH
• protilátky aplikace a dávkování   imunologie   MeSH
• regulace genové exprese u rostlin MeSH
• rekombinantní proteiny aplikace a dávkování   imunologie   MeSH
• signální transdukce MeSH
• umlčování genů imunologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The Francisella tularensis strain LVS phagosome disintegrates during the first few hours after bacterial entry and microbes are released to the cytosol. Within 12 h both rapid multiplication of microbes and a steep increase of apoptosis of infected macrophages occur. We searched for signals involved in the death of macrophages and detected molecules associated with the autophagy machinery cathepsin D, PTEN, p53 and LC3, whose levels or modification were influenced by ongoing in vitro tularemic infection. The sequestration of cytoplasmic F. tularensis LVS into autophagosomes was confirmed by co-localization of the LVS strain containing vacuoles with LC3 (an autophagosomal marker). We also demonstrated the presence of MHC II antigens in these autophagosomes, indicating that they might act as a source of endogenous tularemic antigens for presentation to CD4+ T lymphocytes.

• MeSH
• buněčná smrt genetika   imunologie   MeSH
• cytosol fyziologie   imunologie   mikrobiologie   MeSH
• financování organizované využití   MeSH
• fluorescenční mikroskopie využití   MeSH
• fosfohydroláza PTEN genetika   imunologie   izolace a purifikace   MeSH
• Francisella tularensis imunologie   izolace a purifikace   MeSH
• imunoblotting metody   využití   MeSH
• kathepsin D genetika   imunologie   izolace a purifikace   MeSH
• makrofágy imunologie   mikrobiologie   MeSH
• tularemie etiologie   imunologie   mikrobiologie   MeSH

Autoři vypracují optimální diagnostický a prognostický systém u nemocných s rakovinou prsu za použití nádorových markerů.K vyhodnocení použijí vlastní počítačový program. Od navrženého systému očekávají zlepšení kvality, eventuelně i prodloužení života nemocných s karcinomem prsu.

• MeSH
• cytosol imunologie   MeSH
• diagnostické techniky a postupy využití   MeSH
• diferenciální diagnóza MeSH
• nádorové biomarkery klasifikace   diagnostické užití   MeSH
• nádory prsu diagnóza   MeSH
• plošný screening využití   metody   MeSH
• sérologické testy využití   metody   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• radiologie, nukleární medicína a zobrazovací metody
• onkologie
• chemie, klinická chemie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR