European Journal of Haematology

Abstract: The purpose of the present study was to analyse the importance and prognostic value of renal failure in multiple myeloma patients. The frequency and reversibility of renal failure in 775 multiple myeloma patients diagnosed between 1984–86 and 1990–92 in the Nordic countries were studied. Renal failure, defined as plasma creatinine >130μmol/l, was observed in 29% of the cases at the time of diagnosis. During the first year after diagnosis 58% achieved normalisation of p‐creatinine, and this was achieved mainly during the first 3 months. Reversibility of renal failure was more frequently observed in patients with moderate renal failure, hypercalcaemia and low Bence‐Jones protein excretion. In a multivariate analysis renal failure, high age, stage III disease and hypercalcaemia were independent prognostic factors for survival. Patients who needed dialysis had a poor prognosis, with a median survival of 3.5 months. A 12‐months landmark analysis showed that reversibility of renal failure was a more important prognostic factor than response to chemotherapy. It is concluded that renal failure in multiple myeloma is reversible in about half the cases, and reversibility of renal failure improves long‐term survival.

The physiology of B cells is intimately connected with the function of their B‐cell receptor (BCR). B‐cell lymphomas frequently (dys)regulate BCR signalling and thus take advantage of this pre‐existing pathway for B‐cell proliferation and survival. This has recently been underscored by clinical trials demonstrating that small molecules (fosfamatinib, ibrutinib, idelalisib) inhibiting BCR‐associated kinases (SYK, BTK, PI3K) have an encouraging clinical effect. Here we describe the current knowledge of the specific aspects of BCR signalling in diffuse large B‐cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukaemia (CLL) and normal B cells. Multiple factors can contribute to BCR pathway (dys)regulation in these malignancies and the activation of ‘chronic’ or ‘tonic’ BCR signalling. In lymphoma B cells, the balance of initiation, amplitude and duration of BCR activation can be influenced by a specific immunoglobulin structure, the expression and mutations of adaptor molecules (like GAB1, BLNK, GRB2, CARD11), the activity of kinases (like LYN, SYK, PI3K) or phosphatases (like SHIP‐1, SHP‐1 and PTEN) and levels of microRNAs. We also discuss the crosstalk of BCR with other signalling pathways (NF‐κB, adhesion through integrins, migration and chemokine signalling) to emphasise that the ‘BCR inhibitors’ target multiple pathways interconnected with BCR, which might explain some of their clinical activity.

Tumor necrosis factor (TNF)/cachectin can produce both beneficial and harmful manifestations. Mechanisms may operate to counteract potentially harmful effects such as shock and cachexia. The TNF binding protein (TNF‐BP), which is found at increased levels in serum and urine of patients with chronic renal failure, may play such a role. TNF‐BP was purified 1000000–fold to homogeneity from urine of patients with chronic renal failure by use of ion exchange chromatography, affinity chromatography on TNF‐Sepharose and reverese phase chromatography. The purified protein contained only one chain with an apparent M_r on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of 30000. The aminoterminal amino acid sequence D‐S‐V‐X‐P‐Q‐G‐K‐Y‐I‐H‐P‐Q‐V‐N‐S‐I‐X‐K‐T revealed no significant homologies with previously described protein sequences. TNF‐BP may act as a regulator of the bioactivities of TNF/cachectin.

Abstract: Defensins are a family of small, variably cationic proteins which are highly abundant in the granules of mammalian phagocytes. Three defensins, HNP‐1, 2, and 3, comprise 30–50% of total protein in azurophil granules of human neutrophils. Some defensins are broadly antimicrobial, antiviral and cytotoxic, while others are chemotactic, opsonic, or may modulate hormonal responses. The defensin molecule typically consists of 29–34 amino acids with a conserved pattern of disulfide linkage among its 6 cysteines. The three‐dimensional fold of defensins forms a highly amphiphilic molecule. Microbicidal and cytotoxic properties of defensins are most likely a consequence of their ability to insert into biological membranes and to generate pores. Defensins are synthesized by phagocytes or their precursors as a 94–95 amino acid charge‐neutralized preprodefensin, an arrangement which may avoid cytotoxic injury to the phagocyte. Although defensins were recognized only recently, the existence of homologs in certain invertebrates suggests that they are ancestral components of the host defense system.

Objectives:  MicroRNAs (miRNAs) play key roles in a wide variety of normal and pathological cellular processes. A number of studies identified hematopoietic‐specific miRNAs that are necessary for correct function of blood cells. Out of our microarray data, we chose 13 miRNAs that showed differential expression in peripheral blood cells (miR‐15b, miR‐16, miR‐24, miR‐30c, miR‐106b, miR‐142‐3p, miR‐142‐5p, miR‐150, miR‐155, miR‐181, miR‐223, miR‐342, and miR‐451) and examined their expression in separated hematopoietic cell lineages.

Methods:  Using quantitative real‐time polymerase chain reaction, we measured relative expression of the miRNAs in fractions of reticulocytes, platelets, granulocytes, monocytes, B‐ and T‐lymphocytes as well as in several hematopoietic cell lines.

Results:  We observed that miR‐16 and miR‐142‐3p were highly expressed in all native cell lineages, miR‐451 reached the maximal expression in reticulocytes, miR‐223 in platelets, granulocytes and monocytes, and miR‐150 in B‐ and T‐lymphocytes. Hierarchical clustering analysis grouped the lineage samples according to their origin based on the expression of these miRNAs. To validate discrimination power of the miRNAs, we quantified expression of the 13 miRNAs in several immortalized cell lines. Although the cell lines showed miRNA expression patterns considerably different from those of native cell lineages, clustering analysis distinguished between myeloid, lymphoid and non‐hematopoietic cells.

Conclusions:  In conclusion, the study reports the expression levels of 13 miRNAs in particular blood cell lineages as well as immortalized cell lines. We demonstrate that the expression profiles of these miRNAs may be used for discrimination of the hematopoietic cell lineages.

Abstract: In adult mammals, hemopoiesis takes place primarily in the bone marrow. The steady‐state production of blood cells depends to a large extent on the interaction between hemopoietic stem/progenitor cells (HPC) and the different components of the microenvironment present in the medullary cavity. During the last three decades, in vivo and in vitro studies have allowed significant advances in understanding of the biology of such a hemopoietic microenvironment. Although not evident in histological sections, it is well known that the hemopoietic microenvironment is a highly organized structure that regulates the location and physiology of HPC. The hemopoietic microenvironment is composed of stromal cells (fibroblasts, macrophages, endothelial cells, adipocytes), accessory cells (T lymphocytes, monocytes), and their products (extracellular matrix and cytokines). Microenvironmental cells can regulate hemopoiesis by interacting directly (cell‐to‐cell contact) with HPC and/or by secreting regulatory molecules that influence, in a positive or negative manner, HPC growth. Recent in vitro studies suggest that functional abnormalities of the hemopoietic microenvironment may be implicated in the manifestation of certain hematological disorders such as aplastic anemia, and acute and chronic myelogenous leukemia. Thus, the characterization of the structure and function of the human hemopoietic microenvironment may have relevance in understanding and treating different hematological disorders.

Abstract: The endothelial contribution to vascular disorders has been widely documented in experimental models. However, its implication in human pathology is difficult to investigate, owing to the paucity of non‐invasive methods and of specific endothelial markers. The enumeration of circulating endothelial cells (CEC) released in peripheral blood after vascular injury represents a direct exploration of the endothelium. For this purpose, we have produced a monoclonal antibody (S‐Endo 1), which recognizes CD 146, a molecule expressed on all types of human endothelial cells but absent from haemopoietic cells. Using this antibody, we have designed a specific and sensitive immunocapture test, which allowed us to detect high numbers of CEC in thrombotic, infectious or immunological disorders, while CEC were found to be very rare (<3/ml) in normal subjects. This quantitative approach using CEC might prove useful as a marker of vascular wall injury. Their enumeration is of interest in the clinical follow‐up of vascular disorders, in the evaluation of therapeutic effectiveness or in the direct diagnosis of infectious diseases involving intra‐endothelial microbial agents. Furthermore, an immunological and/or functional study of CEC could allow one to assess their procoagulant and proadhesive properties, as well as their viability, opening new perspectives for CEC investigation in vascular pathology.

Tóm tắt:  Mục tiêu: Đánh giá tình trạng tim mạch ở một nhóm lớn bệnh nhân thalassemia nặng đã được điều trị theo cách tiêu chuẩn từ khi còn nhỏ với liệu pháp truyền máu tích cực và chelat deferoxamine, những người có tuân thủ tốt phác đồ điều trị này. Phương pháp và Kết quả: Chúng tôi đã tiến hành đánh giá lâm sàng và siêu âm tim cho 202 bệnh nhân thalassemia nặng tuổi trung bình 27,3 ± 6,3 năm và 75 đối chứng khỏe mạnh cùng tuổi và giới tính. Bệnh tim mạch biểu hiện rõ đã được phát hiện ở 14 bệnh nhân (6,9%), trong đó có 5 bệnh nhân (2,5%) bị suy tim sung huyết, tuổi từ 26–37 năm, và 9 bệnh nhân bị rối loạn chức năng tâm thu thất trái (LV), tuổi từ 23–37 năm. Mười bệnh nhân (5,0%) có tiền sử viêm màng ngoài tim. Đường kính tâm nhĩ trái và LV, khối lượng LV và lưu lượng tim đều cao hơn đáng kể ở bệnh nhân so với các đối chứng, trong khi kháng lực ngoại vi và gánh nặng thất trái (afterload) thấp hơn đáng kể. Độ dày thành LV tương đối không khác nhau giữa bệnh nhân và đối chứng, nhưng giảm đáng kể ở các bệnh nhân có bệnh tim mạch biểu hiện rõ so với những người không có (P < 0,05). Sự lấp đầy LV có tính hạn chế được quan sát thấy ở 37,6% bệnh nhân và thường gặp hơn ở những trường hợp có bệnh tim mạch biểu hiện rõ (P < 0,01). Tăng huyết áp phổi hầu như không có. Các thông số huyết học và mức áp lực động mạch phổi không có liên quan độc lập với sự hiện diện của bệnh tim mạch biểu hiện rõ. Kết luận: Tuân thủ nghiêm ngặt phác đồ truyền máu và liệu pháp deferoxamine suốt đời đáng kể giúp giảm nguy cơ suy tim, rối loạn chức năng LV và viêm màng ngoài tim, ngăn ngừa suy tim sớm và tăng huyết áp phổi, nhưng không loại bỏ hoàn toàn bệnh tim mạch ở bệnh nhân thalassemia nặng.

Abstract: The clinical syndrome of veno‐occlusive disease (VOD) after hemopoietic cell transplantation is characterised by jaundice, painful liver enlargement, and fluid retention with weight gain. Cytoreductive therapy is presumably the primary cause of VOD, but several other agents (and a special susceptibility of the liver) can also play a role in its genesis. The risk of VOD can be predicted before BMT by analysing the presence or absence of the main risk factors. For the diagnosis of VOD most teams worldwide apply the clinical criteria developed by both the Seattle and Baltimore teams. Transjugular liver studies and some biological markers can help establish a correct differential diagnosis. In most cases clinical manifestations improve after several days, but 20–25% of patients could die of VOD. Data regarding whether or not pharmacological prophylactic measures are effective are contradictory. There a few therapeutical approaches directed towards the improvement of venular occlusion; recombinant tissue plasminogen activator and defibrotide can solve some cases of severe VOD.

Objectives:  To evaluate the prognostic value of absolute lymphocyte count (ALC) at diagnosis in patients with diffuse large B‐cell lymphoma (DLBCL).

Methods:  In a large cohort of patients with DLBCL treated with CHOP (n = 119) or RCHOP (n = 102) in our institution, we evaluated the prognostic value of ALC at diagnosis with regards to treatment response, overall (OS) and progression‐free survival (PFS). Use of rituximab, all International Prognostic Index (IPI) determinants, β2microglobulin level, presence of B symptoms or bulky disease, and ALC were evaluated.

Results:  Low ALC (<1.0 × 10⁹/L) was associated with advanced stage, performance status ≥2, elevated lactate dehydrogenase, number of extranodal involvement ≥2, B symptoms, elevated β2microglobulin and higher IPI risk group. Low ALC was associated with lower CR rate by univariate analysis (odds ratio = 3.29, P = 0.024) but not by multivariate analysis. By univariate analysis using Cox proportional hazard model, low ALC was associated with shorter OS [hazard ratio (HR) = 2.89, P < 0.001] and PFS (HR = 2.91, P < 0.001). Multivariate analysis revealed that low ALC was associated with shorter OS (HR = 2.51, P = 0.003) and PFS (HR = 2.72, P < 0.001), independent of above‐mentioned parameters. Subclass analyses revealed that the use of rituximab improves OS in patients with low ALC (HR = 0.42, P = 0.05) but not in those with high ALC (HR = 0.83, P = 0.71). This observation was most obvious in patients with higher IPI score.

Conclusion:  Low ALC is a poor prognostic marker in patients with DLBCL and suggests patients’ survival benefit from rituximab.