BACKGROUND: Aberrant epigenetic patterns are a hallmark of acute myeloid leukemia (AML). Mutations in profound epigenetic regulators DNMT3A and IDH1/2 often occur concurrently in AML. OBJECTIVES: The aim was to analyze DNA methylation, hydroxymethylation and mRNA expression profiles in AML with mutations in DNMT3A and IDH1/2 (individually and in combinations). METHODS: Infinium MethylationEPIC BeadChip (Illumina) covering 850,000 CpGs was utilized. The validation of hydroxy-/methylation data was done by pyrosequencing. HumanHT-12 v4 Expression BeadChip (Illumina) was used for expression examination. RESULTS: Hierarchical clustering analysis of DNA hydroxy-/methylation data revealed clusters corresponding to DNMT3A and IDH1/2 mutations and CD34+ healthy controls. Samples with concurrent presence of DNMT3A and IDH1/2 mutations displayed mixed DNA hydroxy-/methylation profile with preferential clustering to healthy controls. Numbers and levels of DNA hydroxymethylation were low. Uniformly hypermethylated loci in AML patients with IDH1/2 mutations were enriched for immune response and apoptosis related genes, among which hypermethylation of granzyme B (GZMB) was found to be associated with inferior overall survival of AML patients (P= 0.035). CONCLUSIONS: Distinct molecular background results in specific DNA hydroxy-/methylation profiles in AML. Site-specific DNA hydroxymethylation changes are much less frequent in AML pathogenesis compared to DNA methylation. Methylation levels of enhancer located upstream GZMB gene might contribute to AML prognostication models.

• Klíčová slova
• AML, CHFR, DNMT3A and IDH1/2 mutations, GZMB, hydroxymethylation, methylation,
• MeSH
• akutní myeloidní leukemie genetika   metabolismus   MeSH
• DNA methyltransferasa 3A MeSH
• DNA-(cytosin-5-)methyltransferasa genetika   MeSH
• granzymy genetika   MeSH
• isocitrátdehydrogenasa genetika   MeSH
• leukocyty mononukleární metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• metylace DNA * MeSH
• mutace MeSH
• prognóza MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA methyltransferasa 3A MeSH
• DNA-(cytosin-5-)methyltransferasa MeSH
• DNMT3A protein, human MeSH Prohlížeč
• granzymy MeSH
• GZMB protein, human MeSH Prohlížeč
• IDH1 protein, human MeSH Prohlížeč
• IDH2 protein, human MeSH Prohlížeč
• isocitrátdehydrogenasa MeSH

BACKGROUND: Induction therapy can improve kidney transplantation (KTx) outcomes, but little is known about the mechanisms underlying its effects. METHODS: The mRNA levels of T cell-related genes associated with tolerance or rejection (CD247, GZMB, PRF1, FOXP3, MAN1A1, TCAIM, and TLR5) and lymphocyte subpopulations were monitored prospectively in the peripheral blood of 60 kidney transplant recipients before and 7, 14, 21, 28, 60, 90 days, 6 months, and 12 months after KTx. Patients were treated with calcineurin inhibitor-based triple immunosuppression and induction with rabbit anti-thymocyte globulin (rATG, n = 24), basiliximab (n = 17), or without induction (no-induction, n = 19). A generalized linear mixed model with gamma distribution for repeated measures, adjusted for rejection, recipient/donor age and delayed graft function, was used for statistical analysis. RESULTS: rATG treatment caused an intense reduction in all T cell type population and natural killer (NK) cells within 7 days, then a slow increase and repopulation was observed. This was also noticed in the expression levels of CD247, FOXP3, GZMB, and PRF1. The basiliximab group exhibited higher CD247, GZMB, FOXP3 and TCAIM mRNA levels and regulatory T cell (Treg) counts than the no-induction group. The levels of MAN1A1 and TLR5 mRNA expressions were increased, whereas TCAIM decreased in the rATG group as compared with those in the no-induction group. CONCLUSION: The rATG induction therapy was associated with decreased T and NK cell-related transcript levels and with upregulation of two rejection-associated transcripts (MAN1A1 and TLR5) shortly after KTx. Basiliximab treatment was associated with increased absolute number of Treg cells, and increased level of FOXP3 and TCAIM expression.

• MeSH
• antigeny CD3 genetika   MeSH
• antilymfocytární sérum terapeutické užití   MeSH
• basiliximab MeSH
• dospělí MeSH
• exprese genu účinky léků   MeSH
• forkhead transkripční faktory genetika   MeSH
• granzymy genetika   MeSH
• imunologická tolerance účinky léků   genetika   MeSH
• imunosupresiva terapeutické užití   MeSH
• imunosupresivní léčba metody   MeSH
• indukční chemoterapie metody   MeSH
• inhibitory kalcineurinu terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mannosidasy genetika   MeSH
• messenger RNA krev   MeSH
• mladý dospělý MeSH
• monoklonální protilátky terapeutické užití   MeSH
• NKT buňky účinky léků   imunologie   MeSH
• perforin genetika   MeSH
• počet lymfocytů MeSH
• prospektivní studie MeSH
• regulační T-lymfocyty účinky léků   imunologie   MeSH
• rejekce štěpu genetika   imunologie   prevence a kontrola   MeSH
• rekombinantní fúzní proteiny terapeutické užití   MeSH
• senioři MeSH
• transplantace ledvin metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD3 MeSH
• antilymfocytární sérum MeSH
• basiliximab MeSH
• CD3 antigen, zeta chain MeSH Prohlížeč
• forkhead transkripční faktory MeSH
• FOXP3 protein, human MeSH Prohlížeč
• granzymy MeSH
• GZMB protein, human MeSH Prohlížeč
• imunosupresiva MeSH
• inhibitory kalcineurinu MeSH
• mannosidasy MeSH
• mannosyl-oligosaccharide 1,2-alpha-mannosidase MeSH Prohlížeč
• messenger RNA MeSH
• monoklonální protilátky MeSH
• perforin MeSH
• PRF1 protein, human MeSH Prohlížeč
• rekombinantní fúzní proteiny MeSH

Mucosal-associated invariant T (MAIT) cells contain two main subpopulations, CD8(+) and double-negative (DN) cells. The first reports suggested that subpopulations of MAIT cells have similar phenotype and function. Recent works, however, demonstrate that the subpopulations have different ontogenesis and are differentially affected by xenobiotic treatment. In this work, we re-examined the possible differences between subpopulations of MAIT cells. We demonstrate that the main subpopulations of MAIT cells (CD8 and DN) are relatively uniform in terms of both phenotype and function. Both populations are memory/activated, tissue-homing and pro-inflammatory. CD8(+) MAIT cells are better equipped for pro-inflammatory functions as they express higher levels of CD16 and NKG2D, produce more pro-inflammatory cytokines (TNF-α and IFN-γ) and have higher cytotoxic potential (contain more granzyme B and express higher levels of CD107A upon stimulation). Our study contributes to the understanding of the heterogeneity of MAIT cell population.

• MeSH
• biologické markery metabolismus   MeSH
• buněčný rodokmen imunologie   MeSH
• CD8-pozitivní T-lymfocyty cytologie   imunologie   MeSH
• dospělí MeSH
• exprese genu MeSH
• GPI-vázané proteiny genetika   imunologie   MeSH
• granzymy genetika   imunologie   MeSH
• imunofenotypizace MeSH
• imunologická paměť * MeSH
• interferon gama genetika   imunologie   MeSH
• lektinové receptory NK-buněk - podrodina K genetika   imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• MAIT buňky cytologie   imunologie   MeSH
• membránový protein 1 asociovaný s lyzozomy genetika   imunologie   MeSH
• receptory IgG genetika   imunologie   MeSH
• TNF-alfa genetika   imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• FCGR3B protein, human MeSH Prohlížeč
• GPI-vázané proteiny MeSH
• granzymy MeSH
• GZMB protein, human MeSH Prohlížeč
• IFNG protein, human MeSH Prohlížeč
• interferon gama MeSH
• KLRK1 protein, human MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina K MeSH
• membránový protein 1 asociovaný s lyzozomy MeSH
• receptory IgG MeSH
• TNF-alfa MeSH

The intrinsic apoptosis apparatus plays a significant role in generating and amplifying cell death signals. In this study we examined whether there are differences in the expression of its components and in its functioning in non-small cell lung carcinoma (NSCLC) and the lung. We show that NSCLC cell lines express Apaf-1 and procaspase-9 and -3 proteins and that the expression of Apaf-1 and procaspase-3, but not of procaspase-9 and -7, is frequently up-regulated in NSCLC tissues as compared to the lung. NSCLC tissues and lungs and some NSCLC cell lines expressed also caspase-9S(b) and displayed a high caspase-9S(b)/procaspase-9 expression ratio. Procaspase-3 from NSCLCs and lungs was readily processed to caspase-3 by granzyme B or caspase-8, and the granzyme B-generated caspase-3-like activity was significantly higher in tumor tissues and cells than in lungs. By contrast, cytochrome c plus dATP could induce a significant increase of caspase-3-like activity in cytosol only in some NSCLC cell lines and in subsets of studied NSCLC tissues and lungs, while procaspase-3 and -7 were detectably processed only in NSCLC tissues which showed a high (cytochrome c+dATP)-induced caspase-3-like activity. Taken together, the present study provides evidence that the expression of Apaf-1 and procaspase-3 is up-regulated in NSCLCs and indicates that the tumors have a capability to suppress the apoptosome-driven caspase activation in their cytosol.

• MeSH
• aktiny biosyntéza   MeSH
• aktivace enzymů účinky léků   MeSH
• apoptóza fyziologie   MeSH
• cytochromy c metabolismus   MeSH
• cytosol metabolismus   MeSH
• deoxyadeninnukleotidy metabolismus   MeSH
• dospělí MeSH
• exprese genu MeSH
• faktor 1 aktivující apoptotickou proteasu MeSH
• genetická transkripce MeSH
• granzymy MeSH
• imunoblotting MeSH
• kaspasa 3 MeSH
• kaspasa 7 MeSH
• kaspasy biosyntéza   genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA biosyntéza   genetika   MeSH
• nádorové buněčné linie MeSH
• nádory plic enzymologie   genetika   metabolismus   patologie   MeSH
• nemalobuněčný karcinom plic enzymologie   genetika   metabolismus   patologie   MeSH
• prekurzory enzymů biosyntéza   MeSH
• proteiny genetika   MeSH
• proteosyntéza * MeSH
• senioři MeSH
• serinové endopeptidasy metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• 2'-deoxyadenosine triphosphate MeSH Prohlížeč
• aktiny MeSH
• APAF1 protein, human MeSH Prohlížeč
• CASP3 protein, human MeSH Prohlížeč
• CASP7 protein, human MeSH Prohlížeč
• cytochromy c MeSH
• deoxyadeninnukleotidy MeSH
• faktor 1 aktivující apoptotickou proteasu MeSH
• granzymy MeSH
• GZMB protein, human MeSH Prohlížeč
• kaspasa 3 MeSH
• kaspasa 7 MeSH
• kaspasy MeSH
• messenger RNA MeSH
• prekurzory enzymů MeSH
• proteiny MeSH
• serinové endopeptidasy MeSH

Natural killer cells and cytotoxic T lymphocytes are the main cell populations of the immune system able to directly kill target cells via cytotoxic granules. Different mammalian species may differ in specific features of their pore-forming protein (perforin) and granule-bound serine proteases (granzymes). One perforin gene (PRF1) and four genes encoding granzymes A, B, H, and K (GZMA, GZMB, GZMH, GZMK) were identified in the reference genomes of felids. The objective of this work was to characterize the genes PRF1, GZMA and GZMB in a panel of 17 felid species by next-generation re-sequencing. A search of available felid genomes (17 species) retrieved the coding sequences of these genes for comparison to our data. Both sets of sequences or their combinations (23 species) were used for phylogenetic and selection analyses. Nucleotide PRF1, GZMA and GZMB sequences showed high similarities between felid species (over 95% identity). All trees derived from coding sequences expressed phylogenetic relationships corresponding to the zoological taxonomy of the Felidae, except GZMA. No effects of positive selection were detected in the genes studied, however, effects of purifying selection were observed for PRF1 and GZMA. The conservation of PRF1 is in agreement with its critical biological function. The differentiation observed between granzyme sub-families may reflect an adaptation to pathogen variation. The need to maintain important gene functions and at the same time cope with various pathogens may lead to an equilibrium between positive and negative selective pressures acting on GZMB. The within-species variability in wild felid populations merits further investigation.

• Klíčová slova
• Felidae, NK cell, T cell, cytotoxic granule, granzyme, perforin,
• MeSH
• alely MeSH
• buňky NK * MeSH
• cytotoxické proteiny tvořící póry genetika   MeSH
• cytotoxické T-lymfocyty MeSH
• Felidae * genetika   MeSH
• fylogeneze MeSH
• granzymy genetika   MeSH
• lidé MeSH
• perforin genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytotoxické proteiny tvořící póry MeSH
• granzymy MeSH
• perforin MeSH

T cells are pivotal in the adaptive immune defense, necessitating a delicate balance between robust response against infections and self-tolerance. Their activation involves intricate cross-talk among signaling pathways triggered by the T-cell antigen receptors (TCR) and co-stimulatory or inhibitory receptors. The molecular regulation of these complex signaling networks is still incompletely understood. Here, we identify the adaptor protein ABIN1 as a component of the signaling complexes of GITR and OX40 co-stimulation receptors. T cells lacking ABIN1 are hyper-responsive ex vivo, exhibit enhanced responses to cognate infections, and superior ability to induce experimental autoimmune diabetes in mice. ABIN1 negatively regulates p38 kinase activation and late NF-κB target genes. P38 is at least partially responsible for the upregulation of the key effector proteins IFNG and GZMB in ABIN1-deficient T cells after TCR stimulation. Our findings reveal the intricate role of ABIN1 in T-cell regulation.

• Klíčová slova
• ABIN1, Antigen Receptor, Co-stimulation, T Cells, p38,
• MeSH
• adaptorové proteiny signální transdukční * metabolismus   genetika   MeSH
• aktivace lymfocytů imunologie   genetika   MeSH
• cytotoxické T-lymfocyty * imunologie   metabolismus   MeSH
• diabetes mellitus 1. typu imunologie   genetika   metabolismus   MeSH
• glukokortikoidy indukovaný protein související s TNRF MeSH
• interferon gama metabolismus   MeSH
• lidé MeSH
• mitogenem aktivované proteinkinasy p38 metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• NF-kappa B metabolismus   MeSH
• receptory antigenů T-buněk metabolismus   MeSH
• receptory OX40 metabolismus   genetika   MeSH
• signální transdukce * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adaptorové proteiny signální transdukční * MeSH
• glukokortikoidy indukovaný protein související s TNRF MeSH
• interferon gama MeSH
• mitogenem aktivované proteinkinasy p38 MeSH
• NF-kappa B MeSH
• receptory antigenů T-buněk MeSH
• receptory OX40 MeSH
• Tnfrsf18 protein, mouse MeSH Prohlížeč

Recognition of glycosylation patterns is one of the basic features of innate immunity. Ability of C-type lectin-like receptors such as NKR-P1 to bind saccharide moieties has become recently a controversial issue. In the present study, binding assay with soluble fluorescently labeled recombinant rat NKR-P1A and mouse NKR-P1C proteins revealed apparently no affinity to the various neoglycoproteins. Lack of functional linkage between NKR-P1 and previously described saccharide binder was supported by the fact, that synthetic N-acetyl-D-glucosamine octabranched dendrimer on polyamidoamine scaffold (GN8P) did not change gene expression of NKR-P1 isoforms in C57BL/6 and BALB/c mice divergent in the NK gene complex (both in vitro and in vivo). Surprisingly, N-acetyl-D-glucosamine-coated tetrabranched polyamido-amine dendrimer specifically binds to NKT cells and macrophages but not to NK cells (consistently with changes in cytokine patterns). Despite the fact that GN8P has been tested as an immunomodulator in anti-cancer treatment animal models for many years, surprisingly no changes in cytokine profiles in serum relevant to anti-cancer responses using B16F10 and CT26 harboring mouse strains C57BL/6 and BALB/c are observed. Our results indicate possible indirect involvement of NK cells in GN8P mediated immune responses.

• Klíčová slova
• Anti-tumor immunity, C-type lectin related protein, Carbohydrate dendrimer, Clr, GN4P-A: GlcNAc4-PAMAM-ATTO 565, GN4P-NH(2)-GlcNAc(4)-PAMAM, GN4P: GlcNAc4-PAMAM, GN8P: GlcNAc8-PAMAM, GlcNAc, Gzmb, Macrophages, N-acetyl-d-glucosamine, N-acetyl-d-glucosamine-coated octabranched polyamidoamine dendrimer, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer fluorescently labeled with ATTO 565, N-acetyl-d-glucosamine-coated tetrabranched polyamidoamine dendrimer with free NH(2) group, NK cells, NKG2D, NKR-P1, NKR-P1 receptors, NKT cells, PAMAM dendrimer, PMA, Prf, SBA, SMC, granzyme B, natural killer group 2, member D, natural killer receptor protein 1, perforin, phorbol 12-myristate 13-acetate, polyamidoamine dendrimer, soybean agglutinin, spleen mononuclear cell,
• MeSH
• acetylglukosamin imunologie   metabolismus   MeSH
• buňky NK imunologie   metabolismus   MeSH
• dendrimery metabolismus   MeSH
• experimentální nádory farmakoterapie   genetika   imunologie   MeSH
• exprese genu účinky léků   imunologie   MeSH
• glykokonjugáty imunologie   metabolismus   farmakologie   MeSH
• interferon gama krev   genetika   imunologie   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lektinové receptory NK-buněk - podrodina B genetika   imunologie   metabolismus   MeSH
• lektiny typu C genetika   imunologie   metabolismus   MeSH
• makrofágy imunologie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• NKT buňky imunologie   metabolismus   MeSH
• oligosacharidy imunologie   metabolismus   MeSH
• polyaminy imunologie   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protein - isoformy genetika   imunologie   metabolismus   MeSH
• průtoková cytometrie MeSH
• slezina cytologie   imunologie   metabolismus   MeSH
• TNF-alfa krev   genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acetylglukosamin MeSH
• dendrimery MeSH
• glykokonjugáty MeSH
• interferon gama MeSH
• lektinové receptory NK-buněk - podrodina B MeSH
• lektiny typu C MeSH
• oligosacharidy MeSH
• Poly(amidoamine) MeSH Prohlížeč
• polyaminy MeSH
• protein - isoformy MeSH
• TNF-alfa MeSH