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Leishmaniasis, an intracellular protozoal infection in which tissue macrophages are targeted, is transmitted by female sandfly bite and occurs in 98 countries. Visceral leishmaniasis (VL) is the clinical form of leishmaniasis most frequently associated with HIV, especially in Europe. Both diseases have a synergistic detrimental effect on the cellular immune response. Treatment of VL in patients with underlying HIV-infection is associated with lower cure rates, higher rates of drug toxicity, higher relapse rates and greater mortality than treatment of VL in immunocompetent patients. We report the case of a HIV-1 infected patient with advanced disease who presented VL with multiple relapses. This case highlights the difficulties of treating VL in patients with HIV co-infection.

Constitutively active JAK2V617F and thrombopoietin receptor (TpoR) W515L/K mutants are major determinants of human myeloproliferative neoplasms (MPNs). We show that a TpoRW515 mutation (W515A), whichwedetected in 2 myelofibrosis patients, and the Delta 5TpoR active mutant, where the juxtamembrane R/KW515QFP motif is deleted, induce a myeloproliferative phenotype in mouse bone marrow reconstitution experiments. This phenotype required cytosolic Y112 of the TpoR. Phosphotyrosine immunoprofiling detected phosphorylated cytosolic TpoR Y78 and Y112 in cells expressing TpoRW515A. Mutation of cytosolic Y112 to phenylalanine prevented establishment of the in vivo phenotype and decreased constitutive active signaling by Delta 5TpoR and TpoRW515A, especially via the mitogen-activated protein (MAP)-kinase pathway, without decreasing Janus kinase 2 (JAK2) activation. In contrast, mutation of cytosolic Y78 to phenylalanine enhanced the myeloproliferative syndrome induced by the TpoRW515 mutants, by enhancing receptor-induced JAK2 activation. We propose that TpoR cytosolic phosphorylated Y112 and flanking sequences could become targets for pharmacologic inhibition in MPNs. (Blood. 2010;115:1037-1048)

The Belgian national External Quality Assessment Scheme (EQAS) for haematology organized a survey to assess the reliability of haemoglobin (Hb) measurements with the blood gas analysers (BGAs) currently available in Belgian hospitals. All hospital laboratories received two specimens of fresh EDTA anticoagulated whole blood and were asked to determine the Hb concentration using both the conventional haematology analyser (HA) and all BGAs in the hospital. Ninety-seven hospital laboratories participated in the study and a total of 166 results were reported. The BGAs used (grouped according to technology) were Rapidlab 845, 855, 865 (Bayer 1, n = 41), Rapidlab 1245, 1265, Rapidpoint 405 (Bayer 2, n = 19), GEM Premier 3000 (Instrumentation Laboratory, IL, n = 13), ABL 500 and 600 series (Radiometer 1, n = 13), ABL 700 and 800 series (Radiometer 2, n = 35), Omni C, S5 (Roche 1, n = 7), Omni 3, 6, 9, S2, S4, S6 (Roche 2, n = 21). For the BGAs from Bayer, Radiometer and Roche, interlaboratory variation ranged from 0.6 % to 4.1 %, indicating good precision and close agreement between centres. A significant negative bias observed on the GEM Premier 3000 using the EDTA anticoagulated blood samples did not appear to be present in fresh heparinized whole blood samples. There was no significant difference in imprecision and bias between Hb measurements on BGA situated in and outside the central laboratory.

Objectives: Follicular lymphoma (FL) is the most common adult non-Hodgkin's lymphoma. Diagnosis is based on morphology and can be confirmed by cytogenetic, flow cytometry (FCM) or molecular studies. Despite all these complementary approaches, diagnosis sometimes remains difficult. The purpose of the present work was to characterise the expression of new specific follicular cells markers which allows us to target specifically the abnormal FL cell population in FCM. Methods: A total of 153 samples from healthy subjects and from patients with chronic B-cell lymphoproliferative disorders were analysed by FCM in the same conditions for purpose of comparison. Results: We showed that CD44 is weakly expressed in FL cells compared with peripheral blood mononuclear cell from normal blood donors and others cells from B lymphoproliferative diseases. We nevertheless observed bone marrow samples where some immature B-cell population express CD44 with lower fluorescence intensity. Therefore, we developed a double antibody combination, using CD44 and CD38, which allowed us to separate the normal immature cells from the pathological population using FCM. Conclusion: This new phenotypic approach offers an accurate (sensitivity and specificity of 93% and 96%, respectively), fast and low sample consuming method for the diagnosis of FL.

Whether or not a pregnant women should travel to regions where malaria is highly endemic will always be open to question as no prophlaxis can guarantee complete protection in every case. No chemoprophylaxis is 100% effective or entirely without side-effects, particularly for pregnant women whose immune status provides a favourable environment for the emergence of this parasitosis. The advice given will depend on the country and the region, a possible resistance of the parasite to treatment, the season, the length and circumstances of the stay and individual factors. The choice of treatment is very limited as many therapies are contra-indicated in pregnancy; in addition it is extremely important to give a sufficiently clear explanation concerning precautions and prophylaxis as well as of how a possible bout of malaria should be treated, particularly in case of extended stay.

Whether or not a pregnant women should travel to regions where malaria is highly endemic will always be open to question as no prophlaxis can guarantee complete protection in every case. No chemoprophylaxis is 100 % effective or entirely without side-effects, particularly for pregnant women whose immune status provides a favourable environment for the emergence of this parasitosis. The advice given will depend on the country and the region, a possible resistance of the parasite to treatment, the season, the length and circumstances of the stay and individual factors. The choice of treatment is very limited as many therapies are contra-indicated in pregnancy; in addition it is extremely important to give a sufficiently clear explanation concerning precautions and prophylaxis as well as of how a possible bout of malaria should be treated, particularly in case of extended stay.

Thèse de doctorat en sciences biomédicales (hématologie - biologie moléculaire) (SBIM 3) -- UCL, 2002

Aggressive mastocytosis is a form of systemic mast cell disease (SMCD) characterized by organ infiltration, bone lesions. eosinophilia and lymphadenopathies. Here we report a patient with unusual clinical features, namely osteolysis without other bone lesions commonly found in SMCD, major eosinophilia and cerebral infarction. The mast cells exhibited a classical immunophenotype (CD2+, CD9+, CD13+, CD25+, CD35+, CD45c+ and CD117+). Cytogenetic investigation showed novel complex aberrations, and clonal evolution was correlated with clinical progression. The screening for recurrent point mutations affecting the c-kit gene was negative. Mainly, the ASP816VAL substitution was not detected in our patient. Treatment with steroids and interferon was only temporarily effective.

Digital clubbing with or without hypertrophic osteoarthropathy may occur in a variety of neoplastic diseases. We present the first description of digital clubbing in a patient with progressive mantle cell lymphoma, in the absence of any other apparent etiology. We suggest that clubbing might be a paraneoplastic manifestation in mantle cell lymphoma. The literature on digital clubbing in hematological neoplasms is reviewed.

Because 2-chloro-2'-deoxyadenosine (CdA) is active in B-chronic lymphocytic leukemia (B-CLL), and may interfere with DNA repair, we investigated the potentiating effect of CdA on the cytotoxicity induced in vitro in B-CLL lymphocytes by cyclophosphamide (CP) derivatives, which induce DNA damage by DNA cross-linking. Exposure to CdA at clinically achievable concentrations for 2 h, followed by mafosfamide (MAF) or 4-hydroxycyclophosphamide (4HC) for 22 h, resulted in synergistic cytotoxicity in the majority of B-CLL samples tested. Synergy between CdA and MAF was observed in cell samples of sensitive/untreated patients, as well as in cells of resistant/pretreated patients, particularly at the highest concentrations of MAF. In the cells treated with CdA and MAF, we observed loss in ATP and hallmarks of apoptosis, as evidenced by cellular morphology and high molecular weight DNA fragmentation. The synergy could be explained neither by an influence of MAF on the phosphorylation of CdA, nor by an increase in the incorporation of CdA into DNA in the presence of MAF. The in vitro synergy between CdA and CP derivatives provides a rationale for the use of this association in B-CLL patients.

OBJECT: To examine the hypothesis of Renz and Kalf relative to the involvement of interleukin 1 alpha (IL-1 alpha) in the development of anemia in benzene-exposed workers. According to this hypothesis, benzene inhibits the cleavage of the IL-1 alpha precursor (proIL-1 alpha) to mature IL-1 alpha and the lack of this cytokine is responsible for benzene-induced bone marrow suppression. This inhibition of the processing of proIL-1 alpha is attributed to an inhibition of calpain. METHOD: Selection of a population of mechanics exposed to low levels of benzene from fuels, assessment of usual exposure and lifetime exposure duration, and measurements of concentrations of workplace-air benzene and urinary benzene metabolites. Determination of IL-1 alpha concentrations was done by a whole-blood assay after lipopolysaccharide stimulation and a hematological examination was carried out. Statistical analysis considered several possible confounding factors, particularly smoking and drinking habits. DESIGN: Cross-sectional study. RESULTS: The level of exposure of the mechanics to benzene from fuels was mostly well below 1 ppm. IL-1 alpha production was not decreased in mechanics exposed to benzene from fuels, and no correlation between IL-1 alpha concentrations and red blood cell counts appeared. With the exception of a slight decrease in red blood cell counts in mechanics, no hint of a toxic effect of exposure on hematological parameters was found. CONCLUSIONS: The hypothesis of Renz and Kalf could not be confirmed. Although the low level exposure of the study population and methodological factors are possible explanations, it cannot be excluded that the hypothesis of Renz and Kalf is not generalizable to benzene-exposed humans. Presently, one cannot advise the measurement of IL-1 alpha production for biological effect monitoring of workers exposed to low concentrations of benzene.

The purposes of this study were (a) to determine the prevalence of bone marrow abnormalities in patients with chronic lymphocytic leukemia (CLL) using quantitative MR assessment of axial marrow composition and peripheral marrow distribution; (b) to assess the agreement between both quantitative MR methods and compare their sensitivities to detect marrow alterations; and (c) to correlate MR findings with clinical and laboratory parameters. Twenty-nine consecutive patients with biopsy-proven CLL were investigated on a .5-T MR imager to determine bulk T1 relaxation times of the vertebral bone marrow and proportion of proximal femur surface area occupied by nonfatty marrow on coronal T1-weighted MR images of one hip. Of the 29 patients, 12 (41%) had abnormal increase in lumbar marrow T1 values (>600 msec) and 16 (55%) had increased proportion of surface area occupied by nonfatty marrow in the proximal femur (>+1 SD compared to normal values determined in sex- and age-matched healthy subjects). The results of both quantitative MR methods were normal in 12 patients and abnormal in 11 patients (agreement, 79%). Patients with alterations in peripheral marrow distribution had significantly higher T1 relaxation times (P = .001) than those with normal peripheral marrow. Patients with abnormal marrow composition or distribution at MRI had significantly higher blood and marrow lymphocytosis than patients without these features. In conclusion, the agreement between both quantitative MR methods suggests a parallelism between changes in axial marrow composition and in peripheral marrow distribution in patients with CLL. The limits of quantitative MRI in CLL must be kept in mind, because quantitative MR methods failed to detect leukemic marrow infiltration in 41% of patients.

AIMS: To assess a panel of antibodies on Bouin's fixed bone marrow trephine (BMT) biopsies. These biopsies are widely used in routine diagnosis of various haematological malignancies and may be the sole material available in many centres; however, information regarding the immunostaining of this material is lacking. METHODS: Biopsies were taken from 72 patients presenting with various haematological malignancies (leukaemia, 38; lymphoma, 14; multiple myeloma, 20). A panel of antibodies was assessed on Bouin's fixed BMT biopsies by the alkaline phosphatase-antialkaline phosphatase method. RESULTS: Three B (MB2, LN-2, Ki-B5) and two T cell lineage antibodies (UCHL-1, CD3-r) reliably identified lymphoid cells, while MPO-r, Leu-M1/CD15, and KP-1/CD68 recognised cells from the myeloid or histiocytic/macrophage series. Reed-Sternberg cells were stained by LN-2, Leu-M1, and CD30. Antibodies specific for plasma cells (VS38) and hairy cells (DBA.44) gave a variable pattern of staining. Among the proliferation markers, proliferative cell nuclear antigen but not Ki-67 related antibodies were effective. CONCLUSION: This study presents a panel of antibodies with reactivity not restricted to common fixatives that are also suitable for Bouin's fixed BMT biopsies.

Differential diagnosis of thrombocytosis is not always obvious. The routine clinical chemistry laboratory classically provides only limited help in distinguishing between reactive thrombocytosis (RT) and autonomous thrombocytosis, where platelet production escapes normal regulatory processes, and which is seen in myeloproliferative diseases (MPD) such as essential thrombocythemia and polycythemia vera. We explored the clinical use of platelet distribution width (PDW) in the differential diagnosis of thrombocytosis. During a 3-month period, 250 patients presenting with a platelet count > 500 x 10(9)/L were studied; 174 were classified as having RT, 42 had a diagnosis of MPD, and 34 patients were excluded because they had a hemopathy different from MPD, and either did or did not present a known etiologic factor for RT. First, we determined that in the RT group the value of PDW was closely linked to both mean platelet volume (MPV) and platelet count (PLT) (PDW = 79.5-0.005 PLT -3.5 MPV; r = 0.848, R2 = 0.720). Therefore a new parameter, PDWresidual was defined (PDWresidual = PDWobserved -PDWexpected). Second, the discrimination between reactive and autonomous thrombocytosis obtained with PDWresidual was compared with that obtained with either PDW, MPV, or PLT. PDWresidual provided much more powerful than each of the other parameters used separately: 76% of MPD patients had a PDWresidual above the 95th percentile value of the RT population and none of the MPD patients had a PDWresidual below the 50th percentile. Thus, the combined interpretation of PLT, MPV, and PDW through the use of a PDWresidual appears highly useful in the differential diagnosis of thrombocytosis. Also, through simple modeling, more information can be drawn from parameters such as PDW that hitherto were mostly discarded as being without clinical interest.

The engraftment of human peripheral blood mononuclear cells (Hu-PBMC) from adult donors in scid mice has been published by MOSIER et al. in 1988. The possibility to obtain a secondary human immune response in human-scid mice has also been reported but attempts to induce a primary human immune response still remain difficult to achieve. In this work, an antigen (Canine albumin) or a hapten (DNP) was coupled with tetanus toxoid, an antigenic protein against which our human donors already had memory T cells through vaccination. In this way, hu-scid mice immunized with coupled DNP-tetanus toxoid (TT-DNP) or coupled Canine albumin-Tetanus toxoid (Calb-TT) mounted a specific human immune response anti-DNP or anti-Canine albumin (Calb) respectively. A secondary human immune response anti-tetanus toxoid was also detected in the sera of hu-scid mice immunized with product containing TT but not in the sera of those injected with PBS alone. The scid mice grafted with Hu-PBMC from a TT naive donor and challenged with Calb-TT or Calb alone failed to produce specific anti-Calb antibodies. These observations demonstrate that memory T cells can give a substantial help to naive B cells which interact with them for obvious B cell activation and differentiation into plasma cells. This model of immunization might be useful for other antigens of choice, allowing the production of human monoclonal antibodies, in combination with a suitable system of immortalization. Attempts to immunize human cells in scid mice against DNP coupled to LO-BM2 (a rat monoclonal antibody anti-human IgM) failed to induce a specific human response either anti-rat immunoglobulins (Igs), or anti-DNP and led to a decrease of human Ig production in hu-scid. We also immunized hu-scid mice against ovalbumin alone but, only in some cases, a low specific human immune response was observed, so this system seems to be unreliable.