Cytotoxic T cells and natural killer cells can kill target cells based on their expression and release of perforin, granulysin, and granzymes. Genes encoding these molecules have been only poorly annotated in camelids. Based on bioinformatic analyses of genomic resources, sequences corresponding to perforin, granulysin, and granzymes were identified in genomes of camelids and related ungulate species, and annotation of the corresponding genes was performed. A phylogenetic tree was constructed to study evolutionary relationships between the species analyzed. Re-sequencing of all genes in a panel of 10 dromedaries and 10 domestic Bactrian camels allowed analyzing their individual genetic polymorphisms. The data showed that all extant Old World camelids possess functional genes for two pore-forming proteins (PRF1, GNLY) and six granzymes (GZMA, GZMB, GZMH, GZMK, GZMM, and GZMO). All these genes were represented as single copies in the genome except the GZMH gene exhibiting interspecific differences in the number of loci. High protein sequence similarities with other camelid and ungulate species were observed for GZMK and GZMM. The protein variability in dromedaries and Bactrian camels was rather low, except for GNLY and chymotrypsin-like granzymes (GZMB, GZMH).
- MeSH
- buňky NK metabolismus MeSH
- cytotoxické proteiny tvořící póry genetika MeSH
- cytotoxické T-lymfocyty metabolismus MeSH
- fylogeneze MeSH
- granzymy genetika MeSH
- perforin genetika MeSH
- velbloudovití klasifikace genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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.
- MeSH
- akutní myeloidní leukemie genetika metabolismus 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
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
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
- 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
The granzyme B-induced cell death has been traditionally viewed as a primary mechanism that is used by cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells to eliminate harmful target cells including allogeneic, virally infected and tumour cells. Granzyme B (GrB) is the most abundant serine protease which is stored in secretory granules of CTLs and NK cells. After recognition of the target cell, the engaged CTLs and NK cells vectorially secrete GrB along with other granule proteins including perforin into the immunological synapse. From this submicroscopic intercellular cleft GrB translocates into the cytoplasm of the target cell. Although several models have been proposed to explain the GrB delivery mechanism, conclusive understanding of this process remains still elusive. Once in the cytoplasm, GrB cleaves and activates, or inactivates, multiple protein substrates, resulting eventually into apoptotic demise of the target cell. This review is focused on the gene structure and expression of GrB, its biosynthesis and activation, delivery mechanisms into the target cell cytoplasm, direct proteolytic involvement in activation of several pro-apoptotic pathways, and on regulation of its activity in cancer cells. Moreover, emphasis is given to the GrB-mediated anticancer effects and future clinical applications of the GrB-based and tumour-targeted recombinant fusion constructs.
- MeSH
- aktivace enzymů MeSH
- apoptóza genetika MeSH
- biologické modely MeSH
- granzymy genetika metabolismus fyziologie MeSH
- lidé MeSH
- nádory enzymologie genetika metabolismus MeSH
- regulace genové exprese enzymů MeSH
- regulace genové exprese u nádorů MeSH
- tkáňová distribuce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
PURPOSE: Chimeric transcription factor ETV6/RUNX1 (TEL/AML1) is believed to cause pathologic block in lymphoid cell development via interaction with corepressor complex and histone deacetylase. We wanted to show the regulatory effect of ETV6/RUNX1 and its reversibility by histone deacetylase inhibitors (HDACi), as well as to identify potential ETV6/RUNX1-regulated genes. EXPERIMENTAL DESIGN: We used luciferase assay to show the interaction of ETV6/RUNX1 protein, ETV6/RUNX1-regulated gene, and HDACi. To identify ETV6/RUNX1-regulated genes, we used expression profiling and HDACi in lymphoid cells. Next, using the flow cytometry and quantitative reverse transcription-PCR, we measured differentiation changes in gene and protein expression after HDACi treatment. RESULTS: Luciferase assay showed repression of granzyme B expression by ETV6/RUNX1 protein and the reversibility of this effect by HDACi. Proving this regulatory role of ETV6/RUNX1, we identified, using complex statistical analysis, 25 genes that are potentially regulated by ETV6/RUNX1 protein. In four selected genes with known role in the cell cycle regulation (JunD, ACK1, PDGFRB, and TCF4), we confirmed expression changes after HDACi by quantitative analysis. After HDACi treatment, ETV6/RUNX1-positive cells showed immunophenotype changes resembling differentiation process compared with other leukemic cells (BCR/ABL, ETV6/PDGFRB positive). Moreover, ETV6/RUNX1-positive leukemic cells accumulated in G(1)-G(0) phase after HDACi whereas other B-lineage leukemic cell lines showed rather unspecific changes including induction of apoptosis and decreased proliferation. CONCLUSIONS: Presented data support the hypothesis that HDACi affect ETV6/RUNX1-positive cells via direct interaction with ETV6/RUNX1 protein and that treatment with HDACi may release aberrant transcription activity caused by ETV6/RUNX1 chimeric transcription factor.
- MeSH
- apoptóza účinky záření MeSH
- granzymy genetika MeSH
- histondeacetylasy MeSH
- inhibitory histondeacetylas MeSH
- lidé MeSH
- lymfoidní leukemie genetika patologie MeSH
- lymfopoéza genetika účinky léků MeSH
- nádorové buňky kultivované MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- proliferace buněk účinky léků MeSH
- protein PEBP2A2 fyziologie genetika MeSH
- protoonkogenní proteiny c-ets fyziologie genetika MeSH
- regulace genové exprese u leukemie účinky léků MeSH
- represorové proteiny fyziologie genetika MeSH
- stanovení celkové genové exprese MeSH
- Check Tag
- lidé MeSH