• MeSH
• apoptóza účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• inhibitory histondeacetylas farmakologie   terapeutické užití   MeSH
• klinická studie jako téma MeSH
• kostní dřeň MeSH
• kyselina valproová farmakologie   MeSH
• kyseliny hydroxamové farmakologie   terapeutické užití   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí MeSH
• plazmocelulární leukemie * farmakoterapie   patofyziologie   MeSH
• proliferace buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

OBJECTIVE: A new interleukin-6 (IL-6)-dependent plasma cell leukemia cell line UHKT-944 was established from bone marrow cells derived from a 55-yr-old man with plasma cell leukemia. RESULTS: The cell line possesses phenotypic characteristics of plasma cells including the production of a monoclonal immunoglobulin IgA1-kappa. VH3-9 region of IgVH genes was rearranged and somatically hypermutated. The UHKT-944 cells were found to be negative for most of tested B-cell, T-cell, and myeloid markers. According to cytogenetic analysis, the cells were classified as near tetraploid with several numerical and structural abnormalities including the t(14;20) involving IgH locus. CONCLUSION: The established permanent plasma cell leukemia cell line is a suitable model for the study of cellular and molecular mechanisms of pathogenesis of this rare malignant disease.

• MeSH
• biologické markery MeSH
• cytogenetické vyšetření MeSH
• imunofenotypizace MeSH
• imunoglobuliny genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• plazmocelulární leukemie diagnóza   metabolismus   patologie   MeSH
• proliferace buněk MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH

We established and characterized a new IL-6 dependent multiple myeloma (MM) cell line UHKT-893 from the bone marrow of a relapsed 57-year-old woman. RESULTS: Using nephelometry, cells with plasma cell phenotype and morphology were found to secrete IgG and free kappa (κ)-light chain of immunoglobulin. κ-Light chain was also recognized intracellularly by flow cytometry and by mass spectrometry. VH4-39 region of IgVH genes was rearranged and somatically hypermutated. Cytogenetic analysis of cells revealed new chromosome abnormalities in all breakpoints unique in both MM patients and cell lines - t(1;6), t(1;11), t(5;15), t(5;21), +der(11;15) and der(16). IL-6 independent subline UHKT-893a was established by adaptation to descending IL-6 concentration, while the original cell line keeps on maintaining its IL-6 dependency. CONCLUSION: The cell line provides a suitable material for cellular and molecular studies of tumor abnormalities, with potentially unique mutagenic features of myeloma disease. It may be utilized for human hybridoma construction and vaccine development. Both IL-6 dependent and independent cell clones represent an important model for studies of myeloma cell growth and resistance emerging during targeted therapy.

• MeSH
• buňky kostní dřeně účinky léků   patologie   fyziologie   MeSH
• cytogenetické vyšetření MeSH
• interleukin-6 farmakologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mnohočetný myelom patologie   MeSH
• nádorové buněčné linie MeSH
• primární buněčná kultura * metody   MeSH
• proliferace buněk účinky léků   MeSH
• recidiva MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Vitronectin is an abundant plasma glycoprotein identified also as a part of extracellular matrix. Vitronectin is substantially enriched at sites of injured, fibrosing, inflamed, and tumor tissues where it is believed to be involved in wound healing and tissue remodeling. Little is known about the mechanism of vitronectin localization into the damaged tissues. METHODOLOGY/PRINCIPAL FINDINGS: 2E12 antibody has been described to bind a subset of late apoptotic cells. Using immunoisolation followed by mass spectrometry, we identified the antigen recognized by 2E12 antibody as vitronectin. Based on flow cytometry, we described that vitronectin binds to the late apoptotic and necrotic cells in cell cultures in vitro as well as in murine thymus and spleen in vivo. Confocal microscopy revealed that vitronectin binds to an intracellular cytoplasmic structure after the membrane rupture. CONCLUSIONS/SIGNIFICANCE: We propose that vitronectin could serve as a marker of membrane disruption in necrosis and apoptosis for flow cytometry analysis. Moreover, we suggest that vitronectin binding to dead cells may represent one of the mechanisms of vitronectin incorporation into the injured tissues.

• MeSH
• apoptóza fyziologie   MeSH
• buněčné linie MeSH
• erytrocyty cytologie   metabolismus   MeSH
• hmotnostní spektrometrie MeSH
• Jurkat buňky MeSH
• konfokální mikroskopie MeSH
• kultivované buňky MeSH
• lidé MeSH
• myši MeSH
• nekróza MeSH
• průtoková cytometrie MeSH
• slezina cytologie   metabolismus   MeSH
• thymus cytologie   metabolismus   MeSH
• vazba proteinů MeSH
• vitronektin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Growth-inhibitory and proapoptotic effects of the monoclonal antibody to CD34 molecule, clone 4H11, were tested in CD34+ leukemic cell lines (MOLM-9, JURL-MK1, HEL) and CD34- cell lines (PS-1, ML-2 and CTV-1). We have found that the monoclonal antibody to CD34 inhibited the proliferation and induced apoptosis of all CD34+ cell lines. We did not observe induction of differentiation by anti-CD34 antibody, but a growth arrest of cells in the G0/G1 phase of the cell cycle was detected in all the cell lines studied. Combinations of anti-CD34 antibody with both type I (alpha, beta) or type II (gamma) interferons did not enhance the effects on the cell growth or inhibition of cellular proliferation of the antibody alone. Our data suggest that the monoclonal antibody to CD34 molecule prepared from clone 4H11, after sufficient experimental and preclinical testing on laboratory animals, may provide a new basis for targeted antibody therapy of acute or chronic myeloid leukemia.

Resistance to chemotherapy is one of the major obstacles to effective treatment in acute myeloid leukemia (AML). The most extensively studied protein involved in multidrug resistance (MDR) is the transmembrane glycoprotein P (P-gp), the product of the multidrug resistance gene 1 (MDR1). MDR1/P-gp overexpression is frequently observed in hematological malignancies, especially in acute leukemia, and has been reported to correlate with poor prognosis in acute myeloid leukemia (AML). The aim of this study was to evaluate the level of MDR1 gene expression in bone marrow and/or peripheral blood samples in 92 AML patients in relation to their prognosis. The analyzed group was stratified according to presence or absence of prognostically favorable aberrations (PFAs), such as t(15;17) with PML/RARalpha fusion gene, t(8;21) with AML1/ETO fusion gene or inv(16)/ t(16;16) with CBFbeta/MYH11 fusion gene. These prognostically favorable aberrations were detected by RT-PCR and/or standard cytogenetic techniques. MDR1 expression was detected by semiquantitative comparative RT-PCR using software-based evaluation. The levels of MDR1 expression in the bone marrow predicted induction of complete remission in the whole group of analyzed patients (P = 0.032). They were significantly lower in PFA negative patients who achieved complete remission compared to those who failed to achieve complete remission (P = 0.008). In PFA negative patients, MDR1 expression was higher when compared to PFA positive patients (P = 0.055). No such difference was found when analyzing peripheral blood samples. Our experiments showed no impact of MDR1 expression in bone marrow or peripheral blood cells on overall survival (P = 1.000 and P = 0.903 respectively). In summary, the present study shows the prognostic impact of MDR1 expression on induction of complete remission in AML patients. We confirmed that MDR1 overexpression is an unfavorable prognostic factor in AML, which may help to stratify the risk rate of PFA negative patients. In future studies, quantitative detection of MDR1 expression might be a valuable tool to predict prognosis in this patient subset.

• MeSH
• akutní nemoc MeSH
• amplifikace genu MeSH
• analýza přežití MeSH
• DNA primery MeSH
• financování organizované MeSH
• lidé MeSH
• myeloidní leukemie genetika   mortalita   MeSH
• P-glykoprotein genetika   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protoonkogenní proteiny c-bcr genetika   MeSH
• regulace genové exprese u nádorů MeSH
• Check Tag
• lidé MeSH

Selection and development of new therapeutically useful antibodies for targeted therapy of acute and chronic myeloid leukemia.

Buněčná a molekulární charakterizace nových antireceptorových monoklonálních protilátek proti antigenům leukemických buněk vhodných pro vývoj specifických terapeutik pro léčbu akutní a chronické myeloidní leukemie.

• MeSH
• akutní myelomonocytární leukemie terapie   MeSH
• apoptóza MeSH
• buněčné linie MeSH
• CD antigeny MeSH
• chronická myeloidní leukemie terapie   MeSH
• diferenciační antigeny MeSH
• imunotoxiny MeSH
• monoklonální protilátky MeSH
• myeloidní leukemie terapie   MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• hematologie a transfuzní lékařství
• onkologie
• biologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Early leukemic granulocytic and plasmacytic precursors were studied in vitro and in vivo to provide an information on the intranucleolar distribution of AgNORs (silver stained nucleolus organizer regions). In most of these cells AgNORs appeared as clusters of silver stained particles distributed in the whole nucleolar body. On the other hand, in some leukemic early granulocytic precursors, i.e., in myeloblasts and promyelocytes enlarged AgNORs were translocated in the nucleolar peripheral part. In addition, the number of translocated AgNORs at the nucleolar periphery was significantly smaller. Such translocation of a reduced number of AgNORs was easily produced by experimental aging, i.e., starving of cultured leukemic early granulocytic precursors (HL-60 and K562 cells) in vitro and seems to be reversible. Similar translocation of a reduced number of AgNORs was also produced by aging of leukemic plasmacytic precursors. Thus, the translocation of the reduced number of AgNORs to the nucleolar periphery in some blastic leukemic hematopoietic cells might be an useful marker of their aging at the single cell level. However, more studies in this direction are required in the future.

• MeSH
• barvení stříbrem MeSH
• buněčné jadérko metabolismus   MeSH
• buněčný cyklus fyziologie   MeSH
• buňky K562 MeSH
• buňky kostní dřeně metabolismus   MeSH
• DNA nádorová metabolismus   MeSH
• financování organizované MeSH
• granulocyty metabolismus   ultrastruktura   MeSH
• HL-60 buňky MeSH
• leukemie metabolismus   MeSH
• lidé MeSH
• organizátor jadérka metabolismus   MeSH
• plazmatické buňky metabolismus   ultrastruktura   MeSH
• RNA nádorová metabolismus   MeSH
• stárnutí buněk fyziologie   MeSH
• transport proteinů MeSH
• Check Tag
• lidé MeSH

• Klíčová slova
• maligní plazmatické buňky,
• MeSH
• lidé MeSH
• nádorové buněčné linie * MeSH
• plazmocelulární leukemie MeSH
• plazmocytom MeSH
• Check Tag
• lidé MeSH