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Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and aggressive hematodermic neoplasm usually involving the skin. In this retrospective case series, 10 cases of BPDCN were identified, 90% of which had skin involvement and exhibited predominantly violaceous nodules and/or bruise-like plaques. Skin lesions showed diffuse or nodular dermal-based infiltrates of intermediate sized blasts with a grenz zone. Tumor immunophenotyping was CD4(+), CD56(+), CD123(+) and CD303(+). The most frequently mutated genes according to targeted next-generation sequencing were TET2 (3/7) and NRAS (2/7). Multiagent chemotherapy (CT) was administered as first-line therapy, and a total of 5 patients underwent allogenic hematopoietic stem cell transplantation (allo-HSCT). Better outcomes were observed in younger patients and those treated with acute lymphoblastic leukemia (ALL)-like CT followed by allo-HSCT. This study shows the clinical range of cutaneous lesions of BPDCN. Despite the absence of a gold standard therapy, patients treated with myeloablative intensive regimens and allo-HSCT seems to have a more favorable prognosis., (Copyright © 2024 AEDV. Publicado por Elsevier España, S.L.U. All rights reserved.)

Purpose: To assess the reliability of the differential leucocyte count (DLC) and the left shift flagging (LSF) system provided by the Coulter MAXM (MAXM) haematology analyzer., Material and Methods: 380 blood specimens (drawn with tri-K EDTA as anticoagulant) were studied., Results: By using the reference method (NCCLS H20-A), 50 out of the 380 blood specimens presented abnormal DLC (bands > 6%). Of from these, in 39 (80%) the MAXM displayed LSF of "bands 1 or 2". In 118 left shift flagged specimens (MAXM) with normal manual DLC, 87 (74%) had the "bands 1" alarm and 31 (26%) the "bands 2" alarm. Accordingly if the LSF "bands 1" is overlooked, the percentage of FP decreases from 36% to 10% but the percentage of false negatives (FN) increases from 22% to 58%. In order to improve the appreciation of LSF by decreasing the need of manual revisions, the visual examination of the leucocyte distribution scattergram (LDS), also provided by the MAXM, was conveniently evaluated. This study was performed on 190 blood specimens from which the MAXM displayed a normal DLC in 122 (64%), the LSF of "bands" in 44 (23%) and the LSF of "bands 2" in 24 (12.6%). Of from the 122 specimens with normal DLC, four were FN, of from the 44 specimens with "bands 1" LSF, 37 were FP and of from the 24 specimens with "bands 2" LSF, 16 were FP. The visual appreciation of the LDS showed in the majority of samples with "bands 1" and "bands 2" a definitely different shape consisting in a sharper image up to the top of the picture when compared to samples with normal DLC (without flags). According to this criteria, all the 122 specimens with normal DLC displayed a normal LDS and all the 24 specimens with "bands 2" flag displayed abnormal LDS. Of from the 44 specimens with "bands 1" flag, 26 (59%) showed an abnormal LDS and 18 (41%) a normal LDS. It is noteworthy that of from the 26 specimens with abnormal LDS only 7 were true positive (TP), whereas the 18 specimens with normal LDS all showed a normal DLC according to the reference method. These data allow us to conclude that manual revision was required in 26 out of 68 specimens with "bands 1" and abnormal LDS (13% of the total) and in all the 24 specimens with "bands 2" flag. Therefore by using the information provided by the LDS the need of manual revision decreases to 73% of the total sample with LSF., Conclusion: Our results give further support to the idea that th VCS method used by the Coulter MAXM provides a high quality DLC with specific left shift detection.

Myelodysplastic syndromes (MDS) are a group of acquired hemopathies characterized by peripheral cytopenias due to ineffective hematopoiesis and a high risk of transformation into acute non lymphoblastic leukemia (ANLL) which, in most cases, usually occurs from 6 months to 4 years after diagnosis. A patient with extreme neutropenia with intense dysgranulopoiesis as the only manifestations of MDS is described. The patient was controlled over 14 years and presented multiple infectious episodes, in various locations, throughout the evolution, some being very severe and generally caused by gram-negative germs. Likewise, during this time the patient received different treatments (oxymetholone, prednisone and lithium carbonate) with no hematologic response being observed. The leukocyte count remained around 3 x 10(9)/L with a mean proportion of neutrophils of 12% with no variations being found in the bone marrow aspirates carried out throughout the evolution (total of 9). At 14 years the diagnosis of MDS evolved to ANLL. The patient died shortly after the acute transformation due to respiratory failure secondary to bilateral pneumonia. In this case three peculiar features are of note: the almost exclusive involvement of the granulopoietic series without either anemia or thrombocytopenia, the long evolution of AREB, with acute transformation 14 years after diagnosis and the severity of the infections, among which recurrent lingual granulopenic ulcers were of note.

Purpose: To compare the results of the monocyte count provided by autoanalysers with those attained with flow cytometry., Material and Methods: Fifty-six blood samples (EDTA-K3) from the emergency laboratory were studied. The automatic percentage of monocytes attained by four autoanalysers, STKS (Coulter-IZASA), Technicon H*2 (Bayer Diagnostics), Cell-Dyn-3000 (Abbott Diagnostics) and NE-8000 (TOA-Sysmex) was confronted with that provided by the flow cytometry analysis performed with a FACScan (Becton-Dickinson) using monoclonal antibodies of myeloid-monocytic lineage (CD14, CD16). The morphologic observation of the blood smear in accordance with the H20-A protocol of the National Committee for Clinical Laboratory Standards (NCCLS) was used as a reference method. Descriptive statistics plus linear regression analysis, along with the Student's t test for paired data were used for the statistical evaluation of the results., Results: Significant correlation was found between the systems under study and the flow cytometry method: Technicon H*2 (y = 1.79 +/- 0.61x; r = 0.91, p < 0.0001), Coulter STKS (y = 0.61 +/- 0.93X; r = 0.89, p < 0.0001), Cell-Dyn 3000 (y = 0.88 + 0.64x; r = 0.78, p < 0.0001) and Sysmex NE-8000 (y = 4.60 +/- 0.33x; r = 0.52, p < 0.0001). When comparing the percentage of monocytes by means of the Student's t test it was found that Coulter STKS provided the closest results with regard to flow cytometry (13.78 +/- 9.27 vs 14.19 +/- 8.88, p = 0.49), while the closest findings with respect to the reference method were given by Technicon H*2 (9.29 +/- 4.78 vs 9.71 +/- 4.77, p = 0.30)., Conclusions: From this analysis, it was concluded that flow cytometry could be an alternative method to conventional optic observation in the evaluation of the differential count of leucocytes, including monocytes.

Background: Paroxysmal nocturnal hemoglobinuria (NPH) is a clonal disease in which a deficit in the expression of molecules bound to the cell membrane by glycosyl-phosphatidylinositol (MUGFI) groups has been demonstrated. The MUGFI are widely distributed and among them proteins regulating the action of complement may be found. The development of monoclonal antibodies (MoAb) against MUGFI may allow the introduction of a new diagnostic method in this disease and increase the sensitivity, particularly in transfused individuals., Methods: The erythrocytic and leukocytic phenotype of 14 patients with NPH clonality demonstrated by the classic tests of sensitivity to the complement was analyzed by immunofluorescence techniques and flow cytometry with the use of MoAb which recognize MUGFI (CD55, CD59, CD14, CD16 and CD24)., Results: Cells with a decrease or absence of MUGFI were observed in all the patients. The defect was demonstrated in the red cells, monocytes and neutrophils of all the patients, while it was only observed in the lymphocytes of three patients. The percentage of cells with a decrease in MUGFI was variable (2-100%) as well the pattern of deficiency against the different MoAb used. The MoAb with greatest sensitivity for the detection of clonal population were the CD59 in erythrocytes, the CD14 in monocytes and the CD24 in neutrophils. The CD16 was normal in one patient and the CD55 in two. The transfusion of packed red cells did not influence the abnormal leukocyte pattern, with abnormalities even being observed in the erythrocytary CD59. The Ham test was negative in those cases in which the percentage of negative CD59 erythrocytes was lower than 5% of the total erythrocytic population., Conclusions: The study of glucosylphosphatidylinositol by flow cytometry and monoclonal antibodies is a useful technique for the detection and quantification of the nocturnal paroxistic hemoglobinuria clone even in transfused patients.

Purpose: Splenic lymphoma with circulating villous lymphocytes (SLCVL) is a rare chronic B-type lymphoproliferative disorder, few series having been reported thus far. The major clinical, cytological, immunophenotypic and ultrastructural features, as well as the course and treatment of six cases seen in three hospitals of the Barcelona area in six years are reported herein., Methods: The criteria of Melo et al were followed for the diagnosis of SLCVL. The clinical manifestations at onset and the morphology of lymphocytes from blood and bone-marrow (aspiration and biopsy samples) were analysed. The samples were subjected to cytochemical (acid phosphatase and L-tartaric acid inhibition), immunophenotypic (alkaline immunophosphatase and immunofluorescence) and ultrastructural studies. The histologic study of the spleen of those patients who underwent splenectomy was also performed., Results: The median age of the series was 63 years (range: 45-86). Five of the patients were women. Anaemia was the commonest clinical finding at onset, and splenomegaly was found in all cases. Villous lymphocytes were found in both peripheral blood, 3% to 85%, and bone-marrow, 31% to 70%; they showed diffuse positivity to acid phosphatase stain, this being inhibited in all cases by L-tartaric acid. None of the patients had associated monoclonal gammopathy. Bone-marrow biopsy was performed in five occasions, showing nodular infiltrative pattern in three cases, interstitial in one and diffuse pattern in another. Nodular infiltration was seen in the white pulp of spleen, with no involvement of the red pulp. The ultrastructural study of villous lymphocytes from peripheral blood or bone-marrow disclosed short, slim villi with narrow bases, no lamellar ribosomal complexes being found. Mature B-type lymphocyte proliferation was present in all instances, showing positivity for surface immunoglobulin; monoclonal antibody CD25 was negative in all cases. Three patients underwent splenectomy, and the remainders received no treatment. The expected survival at three years was 80%., Conclusions: SLCVL is a rare B-cell chronic lymphoproliferative disorder with recognised morphologic and immunophenotypic characteristics that make it distinguishable from other B-type lymphoproliferative diseases especially hairy cell leukaemia. Its clinical course is chronic and it has good prognosis. Response to splenectomy is usually favourable, although remission of the disease is not achieved.

A patient with lambda light-chains Bence-Jones multiple myeloma (MM) showed a meningeal myelomatosis during a relapse of his illness. Meningeal infiltration was showed through the detection of plasmatic cells in cerebro spinal fluid, identified morphologic and immunophenotypically, together with hyperproteinemia constituted exclusively by lambda light-chains. Treatment was given, intrathecal (methotrexate and cytosine arabinoside) and systemic (vincristine, adriamycin and dexamethasone) chemotherapy, with disappearance of meningeal infiltration. However the patient died, after three months evolution of MM, tough. Literature on this topic is reviewed.

The Sysmex NE-8000 blood autoanalyser determines the values of red cells and platelets by impedance method and the differential leucocyte count (DLC) by means of radiofrequency plus two independent channels for eosinophils and basophils. The system is provided with a sampler capable of holding 100 closed tubes and its working velocity is about 120 samples per hour. The results of its evaluation are presented in this report. The parameters systematically provided by this cell counter include blood cell counts (plus haemoglobin rate, haematocrit and red cell indices), DLC platelet indices and volumetric distribution curves of red cells and platelets. The machine is provided with alarms on each of the above parameters for any suspicion of pathology. The accuracy analysis was performed with regard to the Technicon H*1 system and the conventional methods recommended by the ICSH. Precision, carry-over, linearity and effective velocity were evaluated in accordance to ICSH standards, and when assessing the DLC the number of visual revisions and the percentage of false positives and negatives were taken into account, and hence the sensitivity, specificity and efficiency of the machine. The accuracy of the parameters analysed was acceptable, except for MCHC and monocyte and eosinophil counts. A fair precision was found except for the monocyte count, along with excellent linearity except for high white-cell count. Less than 1.5% carry-over was observed, except for the leucocyte count. The number of necessary revisions of the DLC was somewhat higher than expected, the false negatives being below 11%. Sensitivity, specificity and efficiency were over 75%, except for hospital patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Reactive leukocytosis has been reported in patients with non-Hodgkin's lymphoma of different histologic types. On the other hand, the blastic crisis of chronic myelocytic leukemia (CML) can sometimes be localized outside the bone marrow and simulate lymphoma, particularly when the blasts are of lymphoid lineage and the blastic crisis is the presenting feature of the disease. We report two patients in whom the differential diagnosis between lymphoblastic lymphoma with reactive leukocytosis and blastic crisis of CML outside the bone marrow was raised. They were two males aged 32 and 22 years, respectively, with lymphadenopathy (and one with splenomegaly), who were initially diagnosed of T lymphoblastic lymphoma. In both cases, leukocytosis was detected with myelemia and dysgranulopoiesis in the onset in one of them and when lymphadenopathy reappeared after remission in the other one. In addition, one patient had marked eosinophilia. In the bone marrow there was marked granulopoietic hyperplasia, with a reduction of fatty cells, and the granulocyte alkaline phosphatase index was reduced. However, the cytogenetic study did not disclose the existence of Philadelphia (Ph) chromosome, and bcr/abl molecular rearrangement was also not observed in the molecular study of both cases. We discuss the basic aspects of differential diagnosis between T lymphoblastic lymphoma with leukemoid reaction and T lymphoid lymphadenopathic blastic crisis of Ph-negative, bcr/abl-negative CML.

The major differences of the autoanalyser Technicon H*2 with regard to the H*1, booth manufactured by the same firm, are the former's higher working speed, over one hundred samples per hour, its hydraulic changes and a new type of samples. This system provides cell counts, leucocyte differential count (LDC) and distribution graphs for cell volume, haemoglobin, platelets and leucocytes. Its accuracy was studied with regard to H*1 and reference methods, along with the precision, linearity, carry-over, necessary LDC revisions and LDC sensitivity, specificity and efficiency. Very good accuracy (r greater than 0.9) was found when except comparing H*2 with the other machine and the reference methods, except for MCHCH and basophil count. The linearity of results was good (r greater than 0.998) and the percentage of carry-over was less than 5% in all cases. Twenty one per cent of hospital patients (HP) and 8% of outpatients (OP) needed revision of LDC. False negative LDC appeared in 0.4% of HP and in 0.3% of OP. LDC sensitivity, specificity in H*2 have solved the problems of H*1, improving the performances and working velocity of the system.

The role played by alpha interferon (alpha-IFN) in the treatment of B-type chronic lymphocytic leukaemia (B-CLL) has been studied by different authors. Despite the inconclusiveness of the results, alpha-IFN seems to be more effective in those patients with low tumour burden (early stages) who have been previously untreated. Although the mechanism of action of alpha-IFN is not wholly understood, it is known that this agent is a strong stimulant of the natural killer lymphocytes (NK). NK activity has been found decreased in B-CLL. In the present work the effect of alpha-IFN on NK activity was studied in 9 previously untreated B-CLL patients in stage A, who received chlorambucil (CLB) followed by alpha-IFN for at least 4 months. The disease stage did not change in most of the patients during alkylating or IFN therapy. CLB failed to increase NK activity, although it diminished the lymphocyte count. Although the lymphocyte count of the patients treated with alpha-IFN was not reduced beyond the values attained by CLB, NK activity reached normal values in 5 of the 7 patients in whom this was low, and kept within normal ranges in the two patients with normal NK activity. The number of CD57+ lymphocytes (this being the antigen present in NK cells) increased after alpha-IFN treatment, without any changes in the remaining T-lymphocyte (CD2, CD4 and CD8) and NK (CD16 and CD11b) subpopulations. These results show that alpha-IFN enhances NK activity in vivo.