Agnieszka Szymczyk

Assessment of red blood cell distribution width as a prognostic marker in chronic lymphocytic leukemia

Red blood cell distribution width (RDW) is a quantitative measure of the variability in size of circulating erythrocytes. It was recently reported that RDW is a prognostic factor for infection diseases, cardiovascular and pulmonary diseases, as well as some neoplasms. Moreover, RDW is remarkably strong predictor of longevity, including all causes of death, for adults aged 45 years and older. To explain this occurrence it was proposed that persistent IGFs/mTOR signaling is one of the factors that play a role in affecting the RDW and mortality. The above observations induced us to analyze the prognostic role of RDW in chronic lymphocytic leukemia (CLL) being the most frequent type of adult leukemia in Western countries. The obtained results have shown that RDW may be considered as a potential CLL prognostic marker. Elevated RDW level at the moment of diagnosis was associated with advanced disease and presence of other poor prognostic factors. It is also connected with overall survival indicating shorter time in patients with elevated RDW. It is possible that the presently observed correlation between mortality and RDW of the CLL patients is affected by their metabolic (IGF-1/mTOR driven)- rather than chronological- aging. The patients with high level of RDW are expected to have an increased persistent level of IGF-1/mTOR signaling. Within the framework of personalized therapy, these CLL patients therefore would be expected to be more sensitive to the treatment with mTOR inhibitors.

Changes in T-cell subpopulations and cytokine network during early period of ibrutinib therapy in chronic lymphocytic leukemia patients: the significant decrease in T regulatory cells number

B cell receptor (BCR) stimulation signal plays an important role in the pathogenesis of chronic lymphocytic leukemia (CLL), and kinase inhibitors directed toward the BCR pathway are now the promising anti-leukemic drugs. Ibrutinib, a Bruton tyrosine kinase inhibitor, demonstrates promising clinical activity in CLL. It is reported that ibrutinib, additionally to directly targeting leukemic cells, also inhibits the interactions of these cells with T cells, macrophages and accessory cells. Assessment of these mechanisms is important because of their non -direct anti-leukemic effects and to identify possible side effects connected with long-term drug administration. The aim of this study was to assess the in vivo effects of ibrutinib on T-cell subpopulations and cytokine network in CLL. The analysis was performed on a group of 19 patients during first month of ibrutinib therapy. The standard multicolor flow cytometry and cytometric bead array methods were used for assessment of T-cell subsets and cytokines/chemokines, respectively. The data obtained indicates that Ibrutinib treatment results in changes in T-cell subpopulations and cytokine network in CLL patients. Particularly, a significant reduction of T regulatory cells in peripheral blood was observed. By targeting these populations of T cells Ibrutinib can stimulate rejection of tumor cells by the immune system.

Target Therapy in Hematological Malignances: New Monoclonal Antibodies

Apart from radio- and chemotherapy, monoclonal antibodies (MoAbs) represent a new, more selective tool in the treatment of hematological malignancies. MoAbs bind with the specific antigens of the tumors. This interaction is a basis for targeted therapies which exhibit few side effects and significant antitumor activity. This review provides an overview of the functional characteristics of MoAbs, with some examples of their clinical application. The promising results in the treatment of hematological malignancies have led to the more frequent usage of MoAbs in the therapy. Development of MoAbs is a subject of extensive research. They are a promising method of cancer treatment in the future.

Imbalance in circulatory iNKT, Th17 and T regulatory cell frequencies in patients with B‑cell non‑Hodgkin's lymphoma

T cells are important in B-cell non-Hodgkins lymphoma immunity, however the function of T cell subsets, including natural killer (iNKT), T helper (Th)17, and T regulatory cells remains to be elucidated. The present study analyzed the frequencies of iNKT, Th17 and T regulatory cells in the peripheral blood of 41 patients with B-cell non-Hodgkin lymphoma at diagnosis, then during and following immunochemotherapy R-CHOP/R-CVP. At lymphoma diagnosis, iNKT and Th17 frequencies were decreased and T regulatory cell frequencies were increased compared with healthy control group. The Th17 cell percentage was lower in patients with a worse prognosis and at a more advanced clinical stage and in contrast, the percentage of T regulatory cells was increased in patients at advanced stages of lymphoma, compared to earlier stages. There was an increase of iNKT and Th17 cells following R-CHOP/R-CVP therapy. In patients that responded, both prior to and following-treatment, percentages of iNKT and Th17 were higher and T regulatory cells were lower compared with patients with subsequent disease progression. Taken together, the results obtained demonstrated the opposing effects of T cell subsets in B-cell lymphoma immunity, with iNKT and Th17 inhibiting and T regulatory cells enhancing tumor growth. These alterations may be caused by malignant B-cells, however there may also be an axis of inverse feedback between T regulatory cells and their interaction with Th17 and iNKT cells.

Agreement between manual and automated measurements of simple QRS/T angle

The spatial QRS/T angle (QRS/T) has been identified as a strong and independent predictor of adverse cardiac events. QRS/T can be determined from the electrocardiogram (ECG) by matrix transformation methods or formula which uses a combination of net QRS and T-wave amplitudes (QRS/Tsimple). Amplitudes can be measured automatically by using dedicated software (QRS/Tauto) or can be manually measured on a computer screen (QRS/Tmanual). This latter method allows analysis of QRS/T, when digital ECGs are not available. The aim of the study was to determine the agreement in the measurements between automatically derived QRS and T amplitudes and manually measured on the computer screen amplitudes. The relative error of the QRS/T between the two methods was estimated in 73 patients. In the case of QRS/Tmanual the inter-observer as well as intra-observer variability was estimated. The relative error between QRS/Tauto vs. QRS/Tmanual was 3.51%. Inter-observer and intra-observer variability of the QRS/Tmanual was 1.19% and 1.18% respectively. Manual measurement of the QRS/T is reliable, however, the predictive value of this parameter should be tested in clinical trials, before QRS/Tmanual can be considered a useful tool in clinical practice or retrospective studies.

THE TECHNIQUE OF FLOW CYTOMETRY IN DIAGNOSTIC RESEARCH

Flow cytometry is a technology that simultaneously counts and then examines multiple physical and/or chemical characteristics of single particles, usually cells, as they flow in a fluid stream through a beam of light. The properties measured include a particle’s relative size, relative granularity or internal complexity, and relative fluo rescence intensity. These characteristics are determined using an optical-to-electronic coupling system that records how the cell scatters incident laser light and emits fluo rescence. One of the most significant applications is immunophenotyping of cells - the most important tool in diagnosis and monitoring haematological disorders, such as acute and chronic leukemia, lymphoma, monoclonal gammopathy, myelodisplastic and myeloproliferative diseases.

Neutropenia in adults – significant diagnostic issue

Abstract Introduction. Neutropenia, a disorder quite commonly encountered in blood tests, is defined as a decrease in the absolute neutrophil count below 1500/µl. Neutropenia may not be clinically significant, whereas it sometimes indicates serious haematological, infectious or rheumatic diseases. The reduction of the number of neutrocytes below 500/µl is referred to as ’agranulocytosis’. Such decrease in neutrophil count impairs host defense and makes the patient more vulnerable to bacterial and fungal infections, which may lead to life-threatening sepsis. Aim. This review presents the causes of congenital and acquired neutropenia, with particular attention to drug-induced neutropenia, which may occur due to the intake of the broad spectrum of drugs, including over-the-counter drugs. The article also attempts to answer the question of how the neutropenia and agranulocytosis should be diagnosed and treated. Methods. The publication is based on the analysis of the literature (PubMed database). Results. It has to be emphasized that a thorough physical examination and appropriate additional tests make it possible to diagnose a disease that causes neutropenia. This allows for the implementation of appropriate therapeutic procedures, and consequently, leads to avoidance of serious infections.

Assessment of micro RNAs expression in leukemic cells as prognostic markers in chronic lymphocytic leukemia: micro RNAs can predict survival in a course of the disease

Numerous genetic alterations predicting prognosis and clinical outcome are revealed recently in chronic lymphocytic leukemia (CLL). Among them the deregulated expression of micro RNAs that can induce tumor growth, or act as tumor suppressors seem to be of great importance. This study aimed to analyze the possible role of chosen micro RNAs as markers of prognosis in patients with CLL. We assessed the expression of miR-21, miR-34a, miR-181a, miR-199a/b and miR-221 in previously separated leukemic cells with the use of qRQ-PCR technique at the moment of diagnosis. The results were then analyzed in regards to presence of prognostic factors, clinical data and the end points like progression free survival (PFS), time to progression (TP) and overall survival time (OS). We detected significant correlations between expression of the analyzed micro RNAs and CLL prognostic markers particularly as far as miR-221 and miR-181a were concerned. The subsequent analysis revealed that high expression of miR-34a and miR-181a as well as low miR-21 expression indicated longer TTP, while miR-221 was predictor of OS. The obtained results prove the role of micro RNAs as CLL prognostic markers, particularly as factors predicting survival in a course of the disease.

Intraventricular treatment of secondary central nervous system lymphoma – Case study and literature overview

Secondary nervous system lymphoma (SCNSL) is a rare extranodal form of non-Hodgkin lymphoma (NHL). This applies to a particular form of lymphoma that does not originally derive from the central nervous system (CNS); it can be both an isolated form of relapse or a systemic part of disease progression. Due to poor prognosis and a lack of established algorithms of therapeutic procedures, it is a big challenge for physicians from many specializations. In our study, we present an interesting case of a patient with a relapsed form of SCNSL for whom a unique form of treatment was used - intraventricular administration of rituximab and methotrexate.

Hodgkin's variant of Richter's transformation during ibrutinib therapy in a series of CLL patients; the Polish Adult Leukemia Group report (PALG)

This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process, which may lead to differences between this version and the Version of Record. Please cite this article as doi: 10.1111/ejh.13016 This article is protected by copyright. All rights reserved. MRS. ELŻBIETA ISKIERKA-JAŻDŻEWSKA (Orcid ID : 0000-0003-2772-8870)