Cílem tohoto projektu je analýza V?1, V?2 a V?3 gamma-delta (??) populací T-lymfocytů u pacientů s chronickou lymfocytární leukémií (CLL). Lidské ?? T buňky jsou účinnými efektorovými buňkami přirozené imunity podílící se na protinádorovému imunitnímu dozoru. Náše pilotní data ukazují významnou reaktivitu ?? T buněk proti primárním CLL buňkám. Naším cílem je detailní analýza populací ?? T buněk včetně stanovení expanzí, klonality, celogenomového profilování, protinádorové cytotoxicity a interakcí s CLL buňkami. Dále bude provedena analýza unikátní kohorty dárců s monoklonální B-buněčnou lymfocytózou (MBL), která přispěje k objasnění příčin a mechanismů patogeneze CLL onemocnění. MBL je biologicky podobné s CLL a je přijímáno jako její prekurzor. Je velmi důležité, abychom dosáhli lepšího porozumění nejen o ?? cytotoxicitě proti maligním CLL buňkám, ale také o reaktivitě ?? T-buněk proti CLL i přes signalizaci od stromatu kostní dřeně. Tato analýza přinese nové výsledky důležité pro budoucí imunoterapeutické aplikace určené k odstranění residuálních CLL buněk.; This project aims to investigate the V?1, V?2 and V?3 subsets of gamma-delta (??) T cells in patients with chronic lymphocytic leukemia (CLL). Human ?? T cells are potent effector cells of innate immunity involved in anti-tumour immune surveillance. Our pilot in vitro data support the observations of prominent reactivity of ?? T cells against primary CLL cells but the detailed analysis of ?? expansions, clonality and anti-CLL reactivity are currently missing. In addition, a unique cohort of monoclonal B-cell lymphocytosis (MBL) donors will be investigated and will lead to important insights into mechanisms of CLL disease pathogenesis. MBL has been shown to display biological similarities to CLL and is accepted as its precursor state. It is crucial that we achieve a better understanding of not only ?? cytotoxicity against malignant CLL cells, but also the efficacy of ?? T cells on CLL cells despite of signaling from bone marrow stroma that will generate novel results highly relevant for future immunotherapeutic applications designed to eliminate the residual CLL cells.

• MeSH
• biologická terapie MeSH
• chronická lymfatická leukemie terapie   MeSH
• imunoterapie MeSH
• intraepiteliální lymfocyty MeSH
• mezibuněčná komunikace MeSH
• protinádorové látky imunologicky aktivní MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• cytologie, klinická cytologie
• alergologie a imunologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

The universal nine-amino-acid transactivation domains (9aaTADs) have been identified in numerous transcription activators. Here, we identified the conserved 9aaTAD motif in all nine members of the specificity protein (SP) family. Previously, the Sp1 transcription factor has been defined as a glutamine-rich activator. We showed by amino acid substitutions that the glutamine residues are completely dispensable for 9aaTAD function and are not conserved in the SP family. We described the origin and evolutionary history of 9aaTADs. The 9aaTADs of the ancestral Sp2 gene became inactivated in early chordates. We next discovered that an accumulation of valines in 9aaTADs inactivated their transactivation function and enabled their strict conservation during evolution. Subsequently, in chordates, Sp2 has duplicated and created new paralogs, Sp1, Sp3, and Sp4 (the SP1-4 clade). During chordate evolution, the dormancy of the Sp2 activation domain lasted over 100 million years. The dormant but still intact ancestral Sp2 activation domains allowed diversification of the SP1-4 clade into activators and repressors. By valine substitution in the 9aaTADs, Sp1 and Sp3 regained their original activator function found in ancestral lower metazoan sea sponges. Therefore, the vertebrate SP1-4 clade could include both repressors and activators. Furthermore, we identified secondary 9aaTADs in Sp2 introns present from fish to primates, including humans. In the gibbon genome, introns containing 9aaTADs were used as exons, which turned the Sp2 gene into an activator. Similarly, we identified introns containing 9aaTADs used conditionally as exons in the (SP family-unrelated) transcription factor SREBP1, suggesting that the intron-9aaTAD reservoir is a general phenomenon.

• MeSH
• aktivace transkripce MeSH
• duplikace genu MeSH
• fylogeneze MeSH
• introny * genetika   MeSH
• lidé MeSH
• molekulární evoluce * MeSH
• regulace genové exprese * MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie MeSH
• transkripční faktor Sp2 * antagonisté a inhibitory   genetika   metabolismus   MeSH
• valin genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH

In higher metazoans, the nuclear hormone receptors activate transcription trough their specific adaptors, nuclear hormone receptor adaptors NCoA, which are absent in lower metazoans. The Nine amino acid TransActivation Domain, 9aaTAD, was reported for a large number of the transcription activators that recruit general mediators of transcription. In this study, we demonstrated that the 9aaTAD from NHR-49 receptor of nematode C.elegans activates transcription as a small peptide. We showed that the ancient 9aaTAD domains are conserved in the nuclear hormone receptors including human HNF4, RARa, VDR and PPARg. Also their small 9aaTAD peptides effectively activated transcription in absence of the NCoA adaptors. We also showed that adjacent H11 domains in ancient and modern hormone receptors have an inhibitory effect on their 9aaTAD function.

• MeSH
• aktivace transkripce * MeSH
• alfa receptor kyseliny retinové chemie   metabolismus   MeSH
• Caenorhabditis elegans chemie   metabolismus   MeSH
• hepatocytární jaderný faktor 4 chemie   metabolismus   MeSH
• lidé MeSH
• molekulární modely MeSH
• peptidy chemie   metabolismus   MeSH
• PPAR gama chemie   metabolismus   MeSH
• proteinové domény MeSH
• proteiny Caenorhabditis elegans chemie   metabolismus   MeSH
• receptory cytoplazmatické a nukleární chemie   metabolismus   MeSH
• receptory kalcitriolu chemie   metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvenční seřazení MeSH
• signální transdukce MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The Gal4 protein is a well-known prototypic acidic activator that has multiple activation domains. We have previously identified a new activation domain called the nine amino acid transactivation domain (9aaTAD) in Gal4 protein. The family of the 9aaTAD activators currently comprises over 40 members including p53, MLL, E2A and other members of the Gal4 family; Oaf1, Pip2, Pdr1 and Pdr3. In this study, we revised function of all reported Gal4 activation domains. Surprisingly, we found that beside of the activation domain 9aaTAD none of the previously reported activation domains had considerable transactivation potential and were not involved in the activation of transcription. Our results demonstrated that the 9aaTAD domain is the only decisive activation domain in the Gal4 protein. We found that the artificial peptides included in the original Gal4 constructs were results of an unintended consequence of cloning that were responsible for the artificial transcriptional activity. Importantly, the activation domain 9aaTAD, which is the exclusive activation domain in Gal4, is also the central part of a conserved sequence recognized by the inhibitory protein Gal80. We propose a revision of the Gal4 regulation, in which the activation domain 9aaTAD is directly linked to both activation function and Gal80 mediated inhibition.

• MeSH
• aktivace transkripce genetika   fyziologie   MeSH
• DNA vazebné proteiny chemie   genetika   metabolismus   MeSH
• molekulární sekvence - údaje MeSH
• proteinové domény MeSH
• Saccharomyces cerevisiae - proteiny chemie   genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• transkripční faktory chemie   genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH

The family of the Nine amino acid Transactivation Domain, 9aaTAD family, comprises currently over 40 members. The 9aaTAD domains are universally recognized by the transcriptional machinery from yeast to man. We had identified the 9aaTAD domains in the p53, Msn2, Pdr1 and B42 activators by our prediction algorithm. In this study, their competence to activate transcription as small peptides was proven. Not surprisingly, we elicited immense 9aaTAD divergence in hundreds of identified orthologs and numerous examples of the 9aaTAD species' convergence. We found unforeseen similarity of the mammalian p53 with yeast Gal4 9aaTAD domains. Furthermore, we identified artificial 9aaTAD domains generated accidentally by others. From an evolutionary perspective, the observed easiness to generate 9aaTAD transactivation domains indicates the natural advantage for spontaneous generation of transcription factors from DNA binding precursors.

• MeSH
• aktivace transkripce * MeSH
• DNA vazebné proteiny chemie   MeSH
• nádorový supresorový protein p53 genetika   MeSH
• Saccharomyces cerevisiae - proteiny chemie   MeSH
• Saccharomyces cerevisiae genetika   MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• transkripční faktory chemie   MeSH
• Publikační typ
• časopisecké články MeSH

Broad changes in human innate and adaptive immunity are associated with advanced age. The age-related alteration of gene expression was reported for both T and B lymphocytes. We analysed the genome-wide expression profiles (n=20) of naive and whole B cell populations from young and early aged healthy donors under 60 years. We revealed large homogeneity of all analysed genome-wide expression profiles but did not identified any significant gene deregulation between young (30-45 years) and early aged healthy donors (50-60 years). We argue that B cells avoid the aging program on molecular level until 60 years of age. Our results demonstrate the potential of hematopoietic stem cells to generate uncompromised lymphocytes in early elderly. These are very encouraging findings for the general health and the immunity maintenance would not need any intervention to naive B cells. Rather, a suitable immune stimulation in healthy body environment warrants further research into aging of older elderly.

• MeSH
• adaptivní imunita genetika   imunologie   MeSH
• B-lymfocyty imunologie   metabolismus   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• přirozená imunita genetika   imunologie   MeSH
• receptor interleukinu-7 - alfa-podjednotka genetika   imunologie   metabolismus   MeSH
• stanovení celkové genové exprese metody   MeSH
• stárnutí genetika   imunologie   MeSH
• transkriptom genetika   imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH