Recent studies have underscored the importance of gamma-delta (γδ) T cells in mediating potent MHC-unrestricted cytotoxicity in numerous malignancies. Here, we analyzed Vδ1 and Vδ2 γδ T cell subsets in newly diagnosed chronic myeloid leukemia (CML) patients (n = 40) who had initiated tyrosine kinase inhibitor (TKI) therapy including imatinib (n = 22), nilotinib (n = 14) and dasatinib (n = 4). Patient peripheral blood samples were analyzed at diagnosis and monitored prospectively at 3, 6, 12 and 18 months post-TKI. γδ T cells isolated from healthy donors and CML patients were used against K562, LAMA-84 and KYO-1 cell lines and against primary CML cells in cytotoxicity assays. We found large expansions of Vδ1 and Vδ2 T cells in patients at diagnosis compared to age-matched healthy donors (n = 40) (p < 0.0001). The γδ T cell reconstitution in patients on imatinib and also on nilotinib showed significant reductions of Vδ1 T cell and Vδ2 T cell absolute counts at 3 months compared to diagnosis. Importantly, Vδ1 and Vδ2 T absolute cell counts remained at normal levels from 3 months throughout the follow-up. Next, we observed susceptibility to specific lysis of primary CML tumor cells by Vδ1 T cells from healthy donors. Furthermore, we determined inherent cytotoxic reactivity by autologous patients' Vδ1 T lymphocytes against primary CML tumor cells. Finally, the TCR clonality profiles showed in CML patients mostly polyclonal repertoires regardless of the TKI. Our results provide further evidence into γδ T cell antileukemia immunity in CML that might be beneficial for long-term disease control and treatment outcome.
- MeSH
- buněčné linie MeSH
- chronická myeloidní leukemie * farmakoterapie metabolismus MeSH
- imatinib mesylát farmakologie terapeutické užití MeSH
- lidé MeSH
- myeloidní leukemie * metabolismus MeSH
- receptory antigenů T-buněk gama-delta metabolismus MeSH
- T-lymfocyty - podskupiny MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
CCCTC-binding factor (CTCF) can both activate as well as inhibit transcription by forming chromatin loops between regulatory regions and promoters. In this regard, Ctcf binding on non-methylated DNA and its interaction with the Cohesin complex results in differential regulation of the H19/Igf2 locus. Similarly, a role for CTCF has been established in normal hematopoietic development; however its involvement in leukemia remains elusive. Here, we show that Ctcf binds to the imprinting control region of H19/Igf2 in AML blasts. We also demonstrate that Smarca5, which also associates with the Cohesin complex, facilitates Ctcf binding to its target sites on DNA. Furthermore, Smarca5 supports Ctcf functionally and is needed for enhancer-blocking effect at ICR. We next asked whether CTCF and SMARCA5 control the expression of key hematopoiesis regulators. In normally differentiating myeloid cells both CTCF and SMARCA5 together with members of the Cohesin complex are recruited to the SPI1 gene, a key hematopoiesis regulator and leukemia suppressor. Due to DNA methylation, CTCF binding to the SPI1 gene is blocked in AML blasts. Upon AZA-mediated DNA demethylation of human AML blasts, CTCF and SMARCA5 are recruited to the -14.4 Enhancer of SPI1 gene and block its expression. Our data provide new insight into complex SPI1 gene regulation now involving additional key epigenetic factors, CTCF and SMARCA5 that control PU.1 expression at the -14.4 Enhancer.
- MeSH
- adenosintrifosfatasy genetika metabolismus MeSH
- akutní erytroblastická leukemie genetika metabolismus patologie MeSH
- akutní nemoc MeSH
- azacytidin farmakologie MeSH
- buňky K562 MeSH
- chromozomální proteiny, nehistonové genetika metabolismus MeSH
- epigeneze genetická * MeSH
- genomový imprinting MeSH
- HeLa buňky MeSH
- imunoblotting MeSH
- insulinu podobný růstový faktor II genetika metabolismus MeSH
- konfokální mikroskopie MeSH
- lidé MeSH
- metylace DNA účinky léků MeSH
- myeloidní leukemie genetika metabolismus patologie MeSH
- nádorové buněčné linie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- protoonkogenní proteiny genetika metabolismus MeSH
- regulace genové exprese u nádorů MeSH
- represorové proteiny genetika metabolismus MeSH
- RNA dlouhá nekódující genetika metabolismus MeSH
- RNA interference MeSH
- trans-aktivátory genetika metabolismus MeSH
- vazba proteinů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The present study was undertaken to provide more information on the differentiation and maturation of human granulocytes using computer-assisted image RNA densitometry at single-cell level. The bone marrow of patients suffering from chronic phase of chronic myeloid leukemia represents a very convenient model for such measurements because of the satisfactory number of early stages, as well as advanced stages, of the granulocytic cell lineage represented by neutrophils. In contrast to the erythroid cell lineage, similar nucleolar and cytoplasmic RNA density-concentration values were found only in early granulocytic progenitors such as myeloblasts and promyelocytes. In advanced stages of the granulocytic development starting with myelocytes, these cells were characterized by a larger decrease in the cytoplasmic RNA concentration in comparison with that of the nucleoli. Thus, the nucleolar to cytoplasmic RNA concentration ratio in these cells was above 1. On the other hand, it should be pointed out that late differentiation stages of granulocytes, starting with myelocytes, possessed nucleolar bodies (nucleoli without surrounding perinucleolar chromatin) of a markedly reduced size.
- MeSH
- biopsie MeSH
- buněčná diferenciace MeSH
- buněčné jadérko ultrastruktura MeSH
- buněčný rodokmen MeSH
- chronická nemoc MeSH
- cytoplazma metabolismus ultrastruktura MeSH
- denzitometrie MeSH
- lidé MeSH
- myeloidní leukemie krev metabolismus patologie MeSH
- počítačové zpracování obrazu MeSH
- prekurzorové buňky granulocytů metabolismus ultrastruktura MeSH
- RNA analýza MeSH
- velikost částic MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- absorpční fotometrie MeSH
- akutní lymfatická leukemie metabolismus terapie MeSH
- akutní nemoc MeSH
- dexamethason aplikace a dávkování škodlivé účinky MeSH
- dítě MeSH
- kostní denzita MeSH
- lidé MeSH
- lineární modely MeSH
- mladiství MeSH
- mladý dospělý MeSH
- multivariační analýza MeSH
- myeloidní leukemie metabolismus terapie MeSH
- následné studie MeSH
- prednison aplikace a dávkování škodlivé účinky MeSH
- přežívající MeSH
- prospektivní studie MeSH
- protokoly antitumorózní kombinované chemoterapie terapeutické užití MeSH
- transplantace hematopoetických kmenových buněk MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- souhrny MeSH
Dimethyl sulfoxide (DMSO) is a widely used prototypical chemical inducer of cell differentiation. In the present study, the effects of DMSO on susceptibility of human myeloid leukemia U937 cells towards ligation of distinct death receptors (DRs) were investigated. DMSO sensitized cells towards induction of apoptosis by anti-Fas antibody, tumour necrosis factor-alpha or Apo2 ligand/TNF-related apoptosis-inducing ligand (TRAIL). Apart from increasing Fas levels, DMSO did not affect expression of proteins in death signal transduction, such as Bcl-2 family proteins, FADD, caspase-3 and -8, the inhibitor of apoptosis proteins (IAPs) or cFLIP(L). However, DMSO significantly potentiated mitochondrial membrane depolarization, suggesting that this mechanism might be involved in sensitisation of myeloid cells to DR-mediated apoptosis.
- MeSH
- adaptorové proteiny signální transdukční metabolismus MeSH
- antigeny CD95 metabolismus MeSH
- apoptóza účinky záření MeSH
- dimethylsulfoxid farmakologie MeSH
- financování organizované MeSH
- FLIP (buněčný) MeSH
- intracelulární signální peptidy a proteiny MeSH
- kaspasa 3 MeSH
- kaspasa 8 MeSH
- kaspasy metabolismus MeSH
- kryoprotektivní látky farmakologie MeSH
- lidé MeSH
- membránové glykoproteiny metabolismus MeSH
- mitochondriální membrány metabolismus patologie MeSH
- mitochondrie metabolismus patologie MeSH
- myeloidní leukemie metabolismus patologie MeSH
- protein FADD asociující s Fas MeSH
- protein TRAIL MeSH
- proteiny regulující apoptózu metabolismus MeSH
- protoonkogenní proteiny c-bcl-2 metabolismus MeSH
- TNF-alfa metabolismus MeSH
- U937 buňky MeSH
- Check Tag
- lidé MeSH
