Katarzyna Kapelko-Słowik

Circulating sCD138 and some angiogenesis-involved cytokines help to anticipate the disease progression of early-stage B-cell chronic lymphocytic leukemia.

Syndecan-1 (CD138) is a transmembrane heparin sulfate proteoglycan expressed on distinct stages of differentiation of B-lymphoid cells. Its prognostic value in B-cell chronic lymphocytic leukemia (B-CLL) has not been evaluated so far. The serum concentration of sCD138 and some angiogenesis-involved cytokines: vascular endothelial growth factor (VEGF), basis fibroblast growth factor (bFGF), and endostatin were studied in 52 previously untreated patients with B-CLL. We found that bFGF and sCD138 levels were significantly higher in B-CLL patients than in controls. In patients with sCD138 level or endostatin level below the median value the lymphocyte count was higher than in patients with serum level of those cytokines above the median value. In patients with progressive disease bFGF level was significantly higher and sCD138 level significantly lower than in patients with stable one. Moreover, high sCD138 level was associated with longer lymphocyte doubling-free survival, and, on the limit of statistical significance, a high endostatin level was associated with shorter progression-free survival. We conclude that serum sCD138 level is increased in early stage B-CLL patients and may have a positive prognostic value as to the dynamics of the disease.

CD8+CD28− Lymphocytes in Peripheral Blood and Serum Concentrations of Soluble Interleukin 6 Receptor are Increased in Patients with Graves’ Orbitopathy and Correlate with Disease Activity

Background. The extrathyroid, orbital manifestation of Graves’ disease (GD)—Graves’ orbitopathy (GO)—presents a difficult clinical problem. The immunological status of GO patients is still under investigation. The aim of this study was to assess the serum concentration of interleukin 6 (IL-6), soluble interleukin 6 receptor (sIL-6R), and CD8+CD28− lymphocytes in GO patients and to evaluate if these parameters were associated with disease activity. Patients. Thirty-nine patients (29 women and 10 men, aged 24–71, mean 50.18) with newly diagnosed GD were enrolled in the study. Active GO was diagnosed in 20 patients. The control group included 12 healthy individuals. Methods. Serum concentrations of IL-6 and sIL-6R were estimated by ELISA. Percentages of CD8+CD28− lymphocytes in peripheral blood were assessed by flow cytometry. Results. Mean serum IL-6 and sIL-6R concentrations were significantly higher in all GD patients and in GO and non-GO patients than in normal controls. In all GD patients and the non-GO group, serum IL-6 and sIL-6R concentrations were significantly reduced after efficient treatment. In GO patients, only serum sIL-6R concentration was significantly lower after efficient treatment. In all GD patients, the mean percentage of CD8+CD28− lymphocytes was significantly lower after efficient treatment. In GO patients, the mean percentage of CD8+CD28− lymphocytes was significantly higher than in the non-GO group or in normals. Moreover, in the GO group, the mean percentage of CD8+CD28− lymphocytes was significantly lower after treatment. Conclusion. Our results have shown that CD8+CD28− lymphocyte percentage in peripheral blood and serum concentration of sIL-6R are increased in GO patients and correlate with disease activity.

Mycotic Infections of the Eye.

Fungal infections of the eye are an important cause of significant visual loss and blindness in some regions of the world, especially developing countries. Ocular mycoses remain a diagnostic and therapeutic challenge to the ophthalmologist. Corneal infection is the most frequent presentation, but the orbit, eyelids, lacrimal apparatus, conjunctiva, sclera and internal structures of the eye can also be affected. Candida spp., Fusarium spp. and Aspergillus spp. are the most frequently isolated organisms in fungal keratitis and in endophthalmitis. The difficulties posed by ocular mycoses are mainly related to establishing the clinical diagnosis, isolation of the fungal pathogen and effective local treatment, particularly in infections of the cornea. The critical issue in diagnosing fungal infection of the eye is microbiological identification of the etiologic agent in clinical samples. Early diagnosis and prompt treatment allow serious complications, including blindness, to be avoided. Local, systemic and even surgical treatment is applied in the therapy.

Expression of PIM-2 and NF-κB genes is increased in patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) and is associated with complete remission rate and overall survival

INTRODUCTION PIM-2 is a proto-oncogene that encodes for a serine/threonine kinase that interacts with various signaling molecules. PIM-2 is highly expressed in neoplastic tissues and in leukemic and lymphoma cell lines, which is consistent with its role during oncogenic transformation. The nuclear factor kappa B (NF-κB) pathway appears to be deregulated in a variety of tumors, with sustained activity of NF-κB leading to apoptotic resistance in tumor cells. The aim of this study was to investigate whether expression of PIM-2 and NF-κB is altered in acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS One hundred forty-three patients were included: 91 with AML and 52 with ALL, aged 18-84 (median 46.7). Eighty-three patients (51 AML and 32 ALL) reached complete remission (CR). Bone marrow samples were collected at the time of diagnosis. Control samples were obtained from 24 healthy donors. We analyzed PIM-2 and NF-κB expression by RQ-PCR analysis. RESULTS Expression of both PIM-2 and NF-κB in all leukemia patients and subgroups was significantly higher than in controls. AML patients who reached CR expressed PIM-2 and NF-κB at significantly lower levels than did patients with primary resistance to chemotherapy and who did not reach CR (NCR). Survival analysis revealed that in AML patients with higher expression of PIM-2 the overall survival (OS) was significantly shorter than in patients with lower expression. CONCLUSION Our data indicate that PIM-2 and NF-κB gene expression is increased in patients with AML and ALL. Moreover, high PIM-2 expression is associated with CR rate and OS in AML patients.

Increased expression of metalloproteinase-2 and -9 (MMP-2, MMP-9), tissue inhibitor of metalloproteinase-1 and -2 (TIMP-1, TIMP-2), and EMMPRIN (CD147) in multiple myeloma

Introduction: Activity of metalloproteinases (MMP) is controlled both by specific tissue inhibitors (TIMP) and activators (extracellular matrix metalloproteinase inducer, EMMPRIN). There are few data available concerning concentration the bone marrow of MMP-2, MMP-9, TIMP-1, and TIMP-2, or EMMPRIM expression by bone marrow mesenchymal stromal cells (BMSCs) in patients with multiple myeloma (MM). Patients and methods: We studied 40 newly diagnosed, untreated patients: 18 males and 22 females with de novo MM and 11 healthy controls. Bone marrow was collected prior to therapy. BMSCs were derived by culturing bone marrow cells on MesenCult. Protein concentrations were determined in bone marrow plasma and culture supernatants by ELISA. EMMPRIN expression by BMSCs was assessed by flow cytometry. Results: The median concentrations of MMP-9, TIMP-1, and TIMP-2 in both marrow plasma and culture supernatants were significantly higher in MM patients than controls. Conclusion: EMMPRIN expression and ratios MMP-9/TIMP-1 and MMP-2/TIMP-2 were higher in MM patients, our results demonstrate that in MM patients MMP-2 and MMP-9 are secreted in higher amounts and are not balanced by inhibitors.

CD117 (c-kit) Expression on CD34+ Cells Participates in the Cytogenetic Response to Imatinib in Patients with Chronic Myeloid Leukemia in the First Chronic Phase

Background: Chronic myeloid leukemia (CML) biology seemed to be perfectly explored especially at the beginning of the tyrosine kinase inhibitors era. Later years with imatinib and second-generation tyrosine kinase inhibitors showed a variety of resistance mechanisms and it became obvious that the bcr-abl chimeric gene is not the only enemy to fight. Some studies assumed the decreased rate of programmed cell death (apoptotic) to be the primary mechanism by which BCR-ABL affects expansion of the leukemic clone in CML. Therefore, the aim of this study was to investigate the role of c-kit inhibition in treatment response. Methods: Cytogenetic analysis, real-time quantitative reverse-transcriptase polymerase chain reaction, flow-cytometric analysis and imatinib serum level quantification were applied. Results: The percentage of CD34+ cells expressing c-kit (CD117) isolated from bone marrow samples of 54 CML patients treated with standard-dose imatinib was significantly lower among imatinib responders. The fraction of apoptotic CD34+CD117+ cells in this patient group was significantly higher than in nonresponders. Conclusion: To achieve optimal treatment response in CML patients, the elimination of CD34+CD117+ may be necessary through an apoptotic pathway.

Atypical chronic myeloid leukaemia: A case of an orphan disease—A multicenter report by the Polish Adult Leukemia Group

Atypical chronic myeloid leukaemia (aCML) belongs to myelodysplastic/myeloproliferative neoplasms. Because of its rarity and changing diagnostic criteria throughout subsequent classifications, data on aCML are very scarce. Therefore, we at the Polish Adult Leukemia Group performed a nationwide survey on aCML. Eleven biggest Polish centres participated in the study. Altogether, 45 patients were reported, among whom only 18 patients (40%) fulfilled diagnostic criteria. Among misdiagnosed patients, myelodysplastic/myeloproliferative syndrome unclassifiable and chronic myelomonocytic leukaemia were the most frequent diagnoses. Thirteen patients were male, median age 64.6 years (range 40.4‐80.9). The median parameters at diagnosis were as follows: white blood cell count 97 × 109/L (23.8‐342) with immature progenitors amounting at 27.5% (12‐72), haemoglobin 8.6 g/dL (3.9‐14.9), and platelet count 66 × 109/L (34‐833). Cytoreductive treatment was used in all patients, and 2 patients underwent allogeneic hematopoietic stem cell transplantation. The median overall survival was 14.1 months (95% CI, 7.2), with median acute myeloid leukaemia‐free survival of 13.3 months (95% CI, 3.6‐22.6). Cumulative incidence of acute myeloid leukaemia transformation after 1 year in aCML group was 12.5% (95% CI, 0%‐29.6%). To conclude, aCML harbours a poor prognosis. Treatment options are limited, with allogeneic hematopoietic stem cell transplantation being the only curative method at present, although only a minority of patients are transplant eligible. Educational measures are needed to improve the quality of diagnoses.

Elevated PIM2 gene expression is associated with poor survival of patients with acute myeloid leukemia

Abstract The PIM2 gene encodes the serine/threonine kinase involved in cell survival and apoptosis. The aim of the study was to evaluate the expression of the PIM2 gene in acute myeloid leukemia (AML) and to examine its role in apoptosis of the blastic cells. We analyzed the PIM2 expression in 148 patients: 91 with AML, 57 with acute lymphoblastic leukemia and 24 healthy controls by Real-Time PCR and Western blot. Inhibition of the PIM2 gene in human leukemic HL60 cell line was performed with RNAi and apoptosis rate was analyzed. Our results indicate that overexpression of PIM2 in AML is associated with low complete remission rate, high-risk cytogenetics, shorter leukemia-free survival, and event-free survival. Cytometric analysis of HL60/PAC-GFP and HL60/PAC-GFP-shPIM2 cells revealed an increase in the number of apoptotic cells after inhibition of PIM2 gene. In summary, the elevated expression of PIM2 in blastic cells is associated with poor prognosis of AML patients and their resistance to induction therapy.

The Hasford Score May Predict Molecular Response in Chronic Myeloid Leukemia Patients: A Single Institution Experience

The Sokal, Hasford, and EUTOS scores were established in different treatment eras of chronic myeloid leukemia (CML). None of them was reported to predict molecular response. In this single center study we tried to reevaluate the usefulness of three main scores in TKI era. The study group included 88 CML patients in first chronic phase treated initially with standard imatinib dose. All of them achieved major molecular response (MMR) in time points defined by European LeukemiaNet (ELN). 42 patients lost MMR in a median time of 47 months and we found a significant difference in MMR maintenance between intermediate-risk (IR) and low-risk (LR) patients assessed by Hasford score. All 42 patients were switched to second-generation TKI (2G-TKI) treatment. At 18 months of 2G-TKI therapy we have still found a significant difference in BCR-ABL transcript levels and MMR rate between IR and LR groups. We did not find any of the described differences discriminating patients by Sokal or EUTOS score. In this retrospective single center analysis we found Hasford score to be useful in predicting molecular response in first chronic phase of CML patients.

Ascites in the course of plasma cell myeloma complicated by AL-amyloidosis.

A 60-year-old Caucasian male with plasma cell myeloma (PCM) immunoglobulin G (IgG) kappa, International Staging System stage 3, diagnosed 5 months ago, was admitted to the department of hematology due to progression of the disease. He had completed three cycles of chemotherapy comprising bortezomib, thalidomide, and dexamethasone; one cycle comprising vincristine, doxorubicin, and dexamethasone; and two cycles comprising lenalidomide and dexamethasone, without any clinically significant response. Three weeks before visiting the hospital, the patient also started complaining of progressive weakness, impaired respiratory function, and abdominal distension; an abdominal ultrasound at the time revealed hepatosplenomegaly with ascites, most likely associated with portal hypertension and protein disturbance, which initially he tolerated very well. Physical examination revealed crackles over the basal areas of the lungs, an enlarged spleen and liver, ascites (stage 2), and peripheral pitting edema. Bone marrow aspiration revealed that plasmacytes accounted for 58% of all nucleated cells. Laboratory tests revealed the following: serum monoclonal IgG, 88.4 g/L (normal: 8-17) and β2-microglobulin, 26.8 mg/L (normal: 1.09-2.53). An abdominal wall fat pad biopsy was positive for amyloid by Congo red staining; this correlated with elevated B-type natriuretic peptide levels (818.7 pg/mL; normal: 0-125). Peritoneal paracentesis was performed and 650 mL of red fluid was aspirated. Laboratory tests revealed a serum-ascites albumin gradient of 1.1 g/dL, with elevated lactate dehydrogenase. Microscopic examination of slide preparations revealed extensive monotonous infiltration by plasmacytes and plasmablasts with highly atypical nuclei and wide polymorphism; monoclonality (CD38+ CD56+ CD45+ CD138+ cyκ+) was confirmed by immunophenotyping (Figure 1A). Computed tomography of the abdomen and thorax revealed interstitial changes in the lower lobes of the lungs; pathological contrast enhancement of enlarged (up to 16-20 mm in diameter) paraaortic, paratracheal, and mediastinal