Hairy cell leukemia (HCL) and HCL-like disorders have to be distinguished because of their different biology and treatment response. Thus, we conducted a retrospective study on patients with HCL and hairy cell leukemia variant (HCLv) to assess diagnostic algorithms and treatment outcomes in a real-world setting. We analyzed 225 HCL and 26 HCLv patients with median follow-up of 67.9 months (HCL) and 20.1 months (HCLv). Median age at diagnosis was 56.2 (HCL) and 69.5 years (HCLv), male predominance was observed in both groups (76.0% vs. 73.1%). Diagnostics was mostly based on morphological evidence of hairy cells in the peripheral blood and bone marrow. At diagnosis, BRAF V600E mutation was detected in 94.7% of examined HCL patients and in no HCLv patient. Front-line treatment was indicated in 205 (91.1%) HCL and 18 (69.2%) HCLv patients. The majority of HCL patients were administered a cladribine-based regimen (91.2%). Overall response rate (ORR) was higher in cladribine-treated patients compared to those given other treatments (97.7% vs. 81.3%), the same applied with achieving Complete remission (CR) (91.2% vs. 62.5%). HCLv treatment was heterogeneous, but cladribine remained the most frequent option (44.4%) with ORR 81.3% and CR rates 43.8%. Second-line treatment was indicated in 52 HCL and 8 HCLv patients, 25.4% and 44.4% of those treated in first-line. In the whole HCL group, median time to next treatment (TTNT) was not reached and 10-year TTNT was estimated at 74.1%. HCLv patients who underwent first-line treatment had a median TTNT of 56 months. The median overall survival (OS) in HCL patients was not reached compared to HCLv with a median OS of 9.5 years. These data confirm an excellent prognosis for HCL patients treated with cladribine-based therapy. On the contrary, HCLv with its aggressive behavior represents a group of patients in whom novel treatment approaches are needed.
- MeSH
- dospělí MeSH
- kladribin terapeutické užití aplikace a dávkování MeSH
- lidé středního věku MeSH
- lidé MeSH
- následné studie MeSH
- protokoly protinádorové kombinované chemoterapie terapeutické užití MeSH
- protoonkogenní proteiny B-Raf genetika MeSH
- retrospektivní studie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- vlasatobuněčná leukemie * diagnóza farmakoterapie patologie mortalita terapie MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
The in vivo rituximab effects in B cell malignancies are only partially understood. Here we analyzed in a large chronic lymphocytic leukemia (CLL) cohort (n = 80) the inter-patient variability in CLL cell count reduction within the first 24 h of rituximab administration in vivo, and a phenomenon of blood repopulation by malignant cells after anti-CD20 antibody therapy. Larger CLL cell elimination after rituximab infusion was associated with lower pre-therapy CLL cell counts, higher CD20 levels, and the non-exhausted capacity of complement-dependent cytotoxicity (CDC). The absolute amount of cell-surface CD20 molecules (CD20 density x CLL lymphocytosis) was a predictor for complement exhaustion during therapy. We also describe that a highly variable decrease in CLL cell counts at 5 h (88 %-2%) following rituximab infusion is accompanied in most patients by peripheral blood repopulation with CLL cells at 24 h, and in ∼20 % of patients, this resulted in CLL counts higher than before therapy. We provide evidence that CLL cells recrudescence is linked with i) CDC exhaustion, which leads to the formation of an insufficient amount of membrane attack complexes, likely resulting in temporary retention of surviving rituximab-opsonized cells by the mononuclear-phagocyte system (followed by their release back to blood), and ii) CLL cells regression from immune niches (CXCR4dimCD5bright intraclonal subpopulation). Patients with major peripheral blood CLL cell repopulation exhibited a longer time-to-progression after chemoimmunotherapy compared to patients with lower or no repopulation, suggesting chemotherapy vulnerability of CLL cells that repopulate the blood.
- MeSH
- chronická lymfatická leukemie krev farmakoterapie imunologie patologie MeSH
- cytotoxicita imunologická imunologie MeSH
- komplement imunologie MeSH
- lidé MeSH
- následné studie MeSH
- protinádorové látky imunologicky aktivní terapeutické užití MeSH
- rituximab terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
cDNA microarray technology is widely used in various biological and medical disciplines to determine gene expression profiles. Unfortunately, this technology requires a large quantity of input RNA. Since there is an increasing need for more precise analyses of defined cell subpopulations with low cell counts, working protocols using a minimal number of input cells are required. Optimal RNA isolation and its accurate amplification are crucial to the success of these protocols. The HL-60 cell line was used in the search for a suitable protocol that can be used for clinical samples of CD34+ haematopoietic cells obtained from bone marrow. The goal was to discover the best method for isolating and amplifying RNA from a small number of cells. Our evaluation of various methods and kits available in the market revealed that the combination of RNAqueous™ Kit for RNA isolation and the SenseAmp Plus Kit for one-round RNA amplification produced the best results. This article presents a verified protocol describing a reliable and reproducible method for obtaining enough input RNA for microarray experiments when the number of cells is limited.
B-cell chronic lymphocytic leukemia (CLL) is an incurable disease with a highly variable clinical course. A proportion of patients eventually progress to a higher stage of malignancy. A recent association has been observed between the presence of aberrant somatic hypermutations in leukemic cells (hypermutations occurring outside of the immunoglobulin locus) and the transformation to a diffuse large B-cell lymphoma or prolymphocytic leukemia. In this study, we report on the rarely observed blastic transformation in a CLL patient who had previously been shown to harbor aberrant somatic hypermutations in the TP53 tumor-suppressor gene (Mol Immunol 2008;45:1525-29). The enzyme responsible, the activation-induced cytidine deaminase, was still active within the transformation, as evidenced by the ongoing class-switch recombination of cytoplasmic immunoglobulins. The transformation was accompanied by a complete p53 inactivation, as well as complex karyotype changes including prominent amplification of MYCN oncogene. Our case-study supports the view that the aberrant somatic hypermutation is associated with transformation of CLL to a more aggressive malignancy.
- MeSH
- amplifikace genu MeSH
- chronická lymfatická leukemie genetika patologie prevence a kontrola MeSH
- financování organizované MeSH
- jaderné proteiny genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádorový supresorový protein p53 genetika MeSH
- onkogenní proteiny genetika MeSH
- recidiva MeSH
- somatická hypermutace imunoglobulinových genů genetika MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Východisko. Krvetvorné buňky periferní krve jsou preferovaným zdrojem transplantátu při léčbě pacientů s non-hodgkinskými lymfomy. Součástí stimulačního chemoterapeutického režimu je aplikace růstových faktorů. Pro ideální výtěžnost sběru štěpu periferních kmenových buněk je nutný překryv časových oken dosaženého maxima leukocytů a koncentrace CD34+ buněk v periferní krvi. Autoři sledovali účinky dvou růstových faktorů (leridistimu a filgrastimu) u pacientů s non-hodgkinskými lymfomy indikovaných k autologní transplantaci na kinetiku a fenotyp CD34+ buněk. Metody a výsledky. Autoři analyzovali fenotyp subpopulací CD34+ buněk a kinetiku jejich mobilizace v periferní krvi a v produktech leukaferézy metodou flowcytometrie v průběhu stimulace a sběru transplantátu. Statisticky významné rozdíly byly zjištěny mezi jednotlivými stimulačními faktory v expresi antigenů: CD3, CD5 (T-linie), CD56 (NK-linie), CD20 (B-linie), CD38 (p < 0,05) a CD54 (p < 0,01). Nejvýznamnější rozdíl byl pozorován v zastoupení CD34+CD19+ subpopulace (B-linie) (p < 0,001). Závěry. Exprese jednotlivých antigenů na CD34+ subpopulacích se v závislosti na stimulačním faktoru statisticky významně lišila. Kinetika vyplavování CD34+ buněk v průběhu stimulace vzhledem k optimalizaci sběru transplantátu nebyla výhodnější pro leridistim oproti filgrastimu.
Background. Peripheral blood stem cells are the preferred source for transplantation of hematopoiesis in patients with non-Hodgkin's lymphoma. Application of hematopoietic growth factors is a part of the mobilization chemotherapy regimen. Time overlap of the highest leukocyte and CD34+ cell count is required for optimal graft collection. Authors analyzed the effect of two growth factors (leridistim and filgrastim) on the kinetics and phenotype of CD34+ cells in patients with non-Hodgkin's lymphoma indicated for autologous peripheral blood stem cell transplantation. Methods and Results. Authors analyzed phenotype of CD34+ cell subpopulations and their kinetics in peripheral blood and leukapheresis products by flow cytometry during mobilization and graft collection. Statistically significant differences in expression of lineage-committed antigens between growth factors were found (CD3, CD5 – T-lineage, CD56 NK-lineage, CD20 for B-lineage, p < 0.05), as well as for lineage non–specific antigens (CD38, p < 0.05 and CD54, p < 0.01). The most significant divergence was observed between CD34+CD19+ subpopulations of leridistim and filgrastim stimulated blood and graft (p < 0.001). Conclusions. Expression of lineage-committed antigens on CD34+ subpopulations between two growth factors was statistically different. Kinetics of CD34+ cells during mobilization regimen with leridistim was not superior to filgrastim concerning the quality of graft.
- MeSH
- antigeny CD34 diagnostické užití MeSH
- autologní transplantace metody MeSH
- faktor stimulující kolonie granulocytů farmakologie terapeutické užití klasifikace terapeutické užití MeSH
- interleukin-3 terapeutické užití MeSH
- nehodgkinský lymfom imunologie terapie MeSH
- průtoková cytometrie využití MeSH
- rekombinantní fúzní proteiny terapeutické užití MeSH
- transplantace periferních kmenových buněk využití MeSH
- Publikační typ
- srovnávací studie MeSH
