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.

• Klíčová slova
• human plasma cell line, interleukin-6 (IL-6), multiple myeloma, plasma cell, plasma cell leukemia,
• 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
• Názvy látek
• biologické markery MeSH
• imunoglobuliny MeSH

UNLABELLED: 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
• Názvy látek
• interleukin-6 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
• Názvy látek
• vitronektin MeSH

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
• 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
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA nádorová MeSH
• RNA nádorová MeSH

Expression of cell cycle-regulating genes was studied in human myeloid leukemia cell lines ML-1, ML-2 and ML-3 during induction of differentiation in vitro. Myelomonocytic differentiation was induced by phorbol ester (12-o-Tetradecanoyl-phorbol-13-acetate, TPA), tumor necrosis factor alpha (TNFalpha) or interferon gamma (INFgamma), or their combination. Differentiation (with the exception of TNFalpha alone) was accompanied by inhibition of DNA synthesis and cell cycle arrest. Inhibition of proliferation was associated with a decrease in the expression of cdc25A and cdc25B, cdk6 and Ki-67 genes, and with increased p21(Waf1/Cip1) gene expression, as measured by comparative RT-PCR. Expression of the following genes was not changed after induction of differentiation: cyclin A1, cyclin D3, cyclin E1 and p27(Kip1). Surprisingly, cyclin D1 expression was upregulated after induction by TPA, TNFalpha with IFNgamma or BA. Cyclin D2 was upregulated only after induction by BA. The results of the expression of the tested genes obtained by comparative RT-PCR were confirmed by quantitative real-time (RQ) RT-PCR and Western blotting. Quantitative RT-PCR showed as much as a 288-fold increase of cyclin D1 specific mRNA after a 24h induction by TPA. The upregulation of cyclin D1 in differentiating cells seems to be compensated by the upregulation of p21(Waf1/Cip1). These results, besides others, point to a strong correlation between the expression of cyclin D1 and p21(Waf1/Cip1) on the one hand and differentiation on the other hand in human myeloid leukemic cells and reflect a rather complicated network regulating proliferation and differentiation of leukemic cells.

• MeSH
• akutní nemoc MeSH
• antitumorózní látky farmakologie   MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčné dělení účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• cyklin D1 genetika   metabolismus   MeSH
• cykliny genetika   metabolismus   MeSH
• DNA primery chemie   MeSH
• inhibitor p21 cyklin-dependentní kinasy MeSH
• interferon gama farmakologie   MeSH
• karcinogeny farmakologie   MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• myeloidní leukemie metabolismus   patologie   MeSH
• nádorové buňky kultivované MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• tetradekanoylforbolacetát farmakologie   MeSH
• TNF-alfa farmakologie   MeSH
• upregulace MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antitumorózní látky MeSH
• CDKN1A protein, human MeSH Prohlížeč
• cyklin D1 MeSH
• cykliny MeSH
• DNA primery MeSH
• inhibitor p21 cyklin-dependentní kinasy MeSH
• interferon gama MeSH
• karcinogeny MeSH
• messenger RNA MeSH
• tetradekanoylforbolacetát MeSH
• TNF-alfa MeSH

Monoclonal antibody 2E12 was prepared by immunization of mice with fresh cells of chronic myeloid leukemia cell line MOLM-7. A panel of 15 leukemic cell lines (myeloid, promyelocytic, erythroid, B and T lymphoid) and numerous cultured patient's leukemia and myeloma cells were tested for reactivity with 2E12 antibody. A subset of cells in all cell lines and various number of patient's cells cultivated for 10 days or more were 2E12 positive. KG-1 and HL-60 cell lines were treated by camptothecin (CAM) (5 microg/ml, 4 h), washed and further cultivated without CAM. After 24 and 48 h in culture a considerable increase of 2E12 positivity was detected both in KG-1 and HL-60 cells, which well correlated with the increase of APO2.7 positivity and the sub-G1 peaks. The 2E12 positive cells were morphologically the same as cells in PCD, possibly apoptosis. We suggest that the 2E12 antibody detects a strong antigen on apoptotic cells which could be a part of the signaling process for ingestion by phagocytes.

• MeSH
• antigeny nádorové imunologie   MeSH
• antitumorózní látky fytogenní toxicita   MeSH
• apoptóza imunologie   MeSH
• chronická myeloidní leukemie genetika   imunologie   MeSH
• DNA nádorová metabolismus   MeSH
• fluorescenční protilátková technika nepřímá MeSH
• kamptothecin toxicita   MeSH
• lidé MeSH
• mnohočetný myelom imunologie   MeSH
• monoklonální protilátky imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nádorové buňky kultivované imunologie   MeSH
• nekróza MeSH
• propidium metabolismus   MeSH
• průtoková cytometrie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny nádorové MeSH
• antitumorózní látky fytogenní MeSH
• DNA nádorová MeSH
• kamptothecin MeSH
• monoklonální protilátky MeSH
• propidium MeSH