The Use of Multicolor Flow Cytofluorometry in White Blood Cell Differential: HematoFlow Conception

Aim. To develop reference intervals for white blood cell subpopulations in peripheral blood using Cytomics FC500 flow cytometer and CytoDiffTM reagent (Beckman Coulter). Materials & Methods. The trial included the analysis of blood samples of healthy donors (n = 315) using Cytomics FC500 flow cytometer and CytoDiffTM reagent cocktail composed of 6 antibodies in 5 colors (CD36-FITC, CD2-PE, CD294-PE, CD19-ECD, CD16-Cy5, CD45-Cy7) and enabling to count 17 cell populations. Results. The data obtained by means of multicolor flow cytofluorometry included the reference values of white blood cell populations in peripheral blood. In 1 first-time donor a lymphoproliferative disease was detected. It was subsequently confirmed by immunophenotyping; B-cell chronic lymphocytic leukemia was diagnosed. Conclusion. Multicolor flow cytofluorometry using CytoDiffTM is considered to be a new step toward an improved WBC differential evaluation aimed mainly at reducing the volume of blood smear analysis using light microscopy at large laboratories, enhancing objectivity, precision and reproducibility of results. WBC differential extended with the count of lymphocyte and monocyte subpopulations can be regarded as modern donor blood screening to detect changes in the pattern of lymphocyte subpopulations as grounds for further examination of donors

Ibrutinib in the Treatment of Refractory Chronic Lymphocytic Leukemia

Background & Aims. This paper presents the results of the observational study of ibrutinib in patients with chronic lymphocytic leukemia (CLL), conducted in SP Botkin Municipal Clinical Hospital. The main objective was the analysis of complications of ibrutinib and identification of factors, influencing the dosage regimen; the secondary objective was the estimation of the total response to treatment, event-free and overall survival. Materials & Methods. The study included 96 patients with CLL with indications for ibrutinib therapy. The median age was 64,9 years (range 32–91 years), the study population consisted of 69 (72 %) men and 27 (28 %) women. The condition of 25 (26 %) patients according to the ECOG scale was of > 3 points. The disease of stage C were diagnosed in 36 (37 %) patients . Deletion of 17p/TP53 mutations were detected in 29 (33 %) of 87 patients. Seventy patients had refractory CLL. The median of the number of the lines of the previous therapy was 3 (range 1–9). Adverse events were assessed in accordance with the CTCAE criteria, version 4.0; the bleeding severity was evaluated using ITP-specific bleeding score; hematological complications were classified according to the recommendations of IWCLL-2008. Results. Ibrutinib was administered at a dosage of 420 mg per day daily until progression or intolerable toxicity. The median duration of ibrutinib therapy was 10.3 months. Ibrutinib was shown to have moderate toxicity, mostly of grade I or II. The bleeding was the most frequent complication. Of the hematological complications, thrombocytopenia was the most common (35 %); neutropenia grade III) developed in 26 % of patients. The treatment response was assessed in 92 patients. The overall response to treatment was 89 %. Complete remission, partial remission and partial remission with lymphocytosis were achieved in 4 (4 %), 57 (62 %), and 21 (23 %) patients, respectively. The event-free survival and overall survival by the month 10 was 90 % and 91 %, respectively. For this observation period, ECOG status and the number of the lines of therapy prior to ibrutinib had the prognostic value. Conclusion. Ibrutinib was shown to have high efficiency in relapsed/refractory forms of CLL. The nature of the ibrutinib toxicity is fundamentally different from that of the conventional chemotherapy. The frequency of ibrutinib therapy complications and patients’ non-compliance depends on the intensity of the previous treatment of CLL. Despite a short observation period, it can be concluded that ibrutinib had the greatest impact on the patient’s quality of life when administered for the first relapse. The low toxicity of ibrutinib is likely to allow the combination with other antitumor agents