bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of protein C receptor expression in monocytes
Authors: Galligan Leeona , Livingstone Wendy , Volkov Yuri , Hokamp Karsten , Murphy Ciaran , Lawler Mark , Fukudome Kenji , Smith Owen
Published: March 2003

Many sequelae associated with endotoxaemic‐induced shock result from excessive production of the cytokine mediators, tumour necrosis factor alpha (TNF‐α), interleukin 1 (IL‐1) and IL‐6 from lipopolysaccharide (LPS)‐activated monocytes. Protein C (PC)/activated protein C (APC) has potent cytokine‐modifying properties and is protective in animal models and human clinical trials of sepsis. The precise mechanism by which this anti‐inflammatory response is achieved remains unknown; however, the recently described endothelial protein C receptor (EPCR) appears to be essential for this function. The pivotal role that monocytes play in the pathophysiology of septic shock led us to investigate the possible expression of a protein C receptor on the monocyte membrane. We used similarity algorithms to screen human sequence databases for paralogues of the EPCR but found none. However, using reverse transcription–polymerase chain reaction (RT–PCR), we detected an mRNA transcribed in primary human monocytes and THP1 cells that was identical to human EPCR mRNA. We also used immunocytochemical analysis to demonstrate the expression of a protein C receptor on the surface of monocytes encoded by the same gene as EPCR. These results confirm a new member of the protein C pathway involving primary monocytes. Further characterization will be necessary to compare and contrast its biological properties with those of EPCR.
Cholelithiasis and Gilbert's syndrome in homozygous β‐thalassaemia
Authors: Galanello R. , Piras S. , Barella S. , Leoni G. B. , Cipollina M. D. , Perseu L. , Cao A.
Published: March 2003

Cholelithiasis has been reported with a variable incidence in homozygous β‐thalassaemia, the reasons for which have only partially been defined. Disease‐associated factors or specific modifier genes may be implicated. We assessed the prevalence of cholelithiasis and the effect of co‐inherited Gilbert's syndrome genotype on its development in 261 thalassaemia major (TM) and 35 thalassaemia intermedia (TI) patients. Cholelithiasis was found in 20·3% of TM and in 57·1% of TI patients. Its incidence was higher (P < 0·05) in patients homozygous for the (TA7) motif in the promoter of the UGT1‐A1 gene, the genotype associated with Gilbert's syndrome, which seems to be a risk factor for the development of gallstones in TM and TI patients.
Generation of reactive oxygen species is not involved in idarubicin‐induced apoptosis in human leukaemic cells
Authors: Liu Feng‐Ting , Kelsey Stephen M. , Newland Adrian C. , Jia Li
Published: March 2003

The anthracycline antibiotic idarubicin (IDA) induces double‐stranded DNA breaks, the generation of reactive oxygen species (ROS) and apoptosis in human leukaemic cells. It is unclear whether the generation of ROS is associated with the apoptotic process. Using the T‐lymphoblastic leukaemic CEM cell line, we found that IDA‐induced DNA breaks were correlated with final cell death. The reduction in mitochondrial membrane potential (ΔΨm) and the generation of ROS occurred simultaneously with IDA‐induced activation of caspase‐9 and caspase‐3. Inhibition of caspases by a pan‐caspase inhibitor, benzyloxycarbonyl‐Val‐Ala‐Asp‐fluoromethyl ketone (Z‐VAD‐fmk) completely blocked IDA‐induced reduction of ΔΨm, apoptosis and final cell death. Interestingly, ROS generation was significantly enhanced by Z‐VAD‐fmk. ROS generation was neither caspase dependent nor part of the apoptotic process. IDA‐mediated reduction in ΔΨm is caspase dependent and is not a consequence of the generation of ROS. These results indicate that IDA‐induced generation of ROS and apoptosis are separate events. Inhibition of caspases facilitates IDA‐mediated generation of ROS.
Novel alleles at the JK blood group locus explain the absence of the erythrocyte urea transporter in European families*
Authors: Irshaid Nidal M. , Eicher Nicole I. , Hustinx Hein , Poole Joyce , Olsson Martin L.
Published: March 2003

Summary.  The Kidd (JK) blood group system is of importance in transfusion medicine. The Jknull phenotype is associated with absence of the urea transporter in erythrocytes and moderately reduced ability to concentrate urine. We and others recently reported different molecular alterations in the silenced Jkb‐like alleles of Polynesians and Finns, populations with higher Jknull frequencies. Here we report novel molecular bases of this phenotype in Caucasians. Blood samples from a Swiss and an English family were investigated by serological methods, urea haemolysis test and JK genotyping. Genomic DNA and JK mRNA were sequenced. Genotyping showed homozygosity for Jka‐like alleles. The Swiss Jknull alleles deviated from wild‐type Jka sequence by a nonsense mutation in exon 7 causing an immediate stop codon (Tyr194stop). The English Jknull alleles revealed a genomic 1·6 kilobase pair deletion including exons 4 and 5, the former of which includes the translation start codon. Multiple mRNA splicing variants were detected in reticulocytes but exons 3–5 were absent in all transcripts analysed. Screening for these alleles was negative in random donors. Two novel molecular alterations at the JK locus were defined and a multiplex polymerase chain reaction method for detection of the five known silent Jk alleles was developed to complement JK genotyping in clinical transfusion medicine.
Expression of transcription factors Pu.1, Spi‐B, Blimp‐1, BSAP and oct‐2 in normal human plasma cells and in multiple myeloma cells
Authors: Nagy Monika , Chapuis Bernard , Matthes Thomas
Published: March 2003

Summary. Differentiation of B lymphocytes into plasma cells is regulated by the interaction of distinct transcription factors (TFs) which activate gene expression in a lineage‐ and stage‐specific pattern. Using reverse transcription polymerase chain reaction, we studied the expression of five TFs (octamer binding factor oct‐2, ets family members PU.1 and Spi‐B, pax gene family member BSAP, and Blimp‐1) in (1) human cell lines with a plasma cell phenotype, (2) primary malignant plasma cells [obtained from patients with plasma cell leukaemia (PCL) and multiple myeloma], and (3) normal human plasma cells generated in vitro or isolated from normal bone marrows. The expression pattern was compared with TFs expressed by normal CD19+ B lymphocytes and by B cells from chronic lymphocytic leukaemia patients. Our results showed that plasma cells expressed a restricted set of TFs compared with CD19+ B lymphocytes, with continued expression of Spi‐B and oct‐2, increased Blimp‐1 expression, and downregulation of BSAP and PU.1. Cells from PCL lost Spi‐B and PU.1 expression completely and expressed only oct‐2 and Blimp‐1, and thus resembled plasma cell lines. Human plasma cell differentiation therefore seems to be positively regulated by Blimp‐1; whether this TF has any oncogenic potential will have to be analysed in future studies.
Axillary vein thrombosis in a healthy donor following platelet apheresis
Authors: Jones D. A. , Williams E. , Riley S. A. , Makris M.
Published: March 2003

Summary. Platelet donation is a common and usually safe procedure accounting for almost half of the platelets donated in the UK. We describe a previously healthy female donor who developed an axillary vein thrombosis following an apheresis platelet donation.
A novel rearrangement of the human fetal globin genes leading to a six γ‐globin gene haplotype
Authors: Gonçalves Isabel , Lavinha João , Ducrocq Rolande , Osório‐Almeida Leonor
Published: March 2003

Summary. Haematological as well as gene mapping data are reported for three members of a Portuguese Caucasian family with high Gγ‐globin levels. A gamma‐globin gene sextuplication of the GγAGγAGγAGγAGγAγ type was present in the proband and her father. Comparison of gene mapping data with quantitative results of fetal haemoglobin (HbF) analysis provided an explanation for the extremely high Gγ‐globin levels (> 90%) in the HbF from the two mentioned individuals. This rearrangement, for which a generation mechanism is proposed, is the first gamma‐globin gene sextuplication described in the literature.
An estimate of the current risk of transmitting blood‐borne infections through blood transfusion in Italy
Authors: Tosti M. E. , Solinas S. , Prati D. , Salvaneschi L. , Manca M. , Francesconi M. , Ciuffreda M. , Girelli G. , Mele A.
Published: March 2003

Summary.We conducted a retrospective cohort study to estimate the incidence of major blood‐borne agents among Italian blood donors and calculated the risk of infection among blood recipients using the ‘incidence/window period model’. The study was conducted among 46 180 blood donors enrolled in six blood centres between 1994 and 1999. During follow‐up, seven new infections were confirmed: three donors seroconverted for anti‐human immunodeficiency virus (HIV); two for anti‐hepatitis C virus (HCV); and two showed hepatitis B surface antigen (HBsAg) reactivity; no cases of syphilis were observed. The incidence rates per 100 000 person/years were: 4·06 (95% CI: 0·82–11·85) for HIV; 2·41 (95% CI: 0·29–8·70) for HCV; and 2·70 (95% CI: 0·32–9·77) for HBsAg; the incidence for total hepatitis B virus (HBV) infection was 9·77 per 100 000 person/years (95% CI: 1·16–35·36). The estimated risk of an infectious blood unit not being detected was: 2·45 (95% CI: 0·13–12·33) per 1 million units for HIV; 4·35 (95% CI: 0·30–22·39) for HCV; and 15·78 (95% CI: 1·16–84·23) for HBV. Overall, an estimated 22·58 per 1 million units are infected. In Italy, the risk of transfusion‐transmitted infections is low and is similar to that in other western countries. The introduction of new more sensitive screening tests could reduce the residual risk of transfusion‐transmitted infection by 40–80%.
Enhanced antitumoral effectiveness of idiotype vaccination induced by the administration of Flt3 ligand combined with interleukin 2 against a murine myeloma
Authors: Zeis Matthias , Zunkel Tim , Steinmann Jörg , Schmitz Norbert , Uharek Lutz
Published: March 2003

Summary. Idiotype (Id) vaccination provides an innovative treatment modality against B‐cell malignancies. In multiple myeloma patients, however, the antitumoral potential of this immunotherapeutic concept is limited. In an attempt to improve the therapeutic effectiveness of Id vaccination, we added Flt3 ligand (Flt3‐L) and interleukin 2 (IL‐2) to the protocol. Balb/c mice were inoculated i.p. (d −2) with different doses (1–5 × 105) of HOPC myeloma cells, secreting the IgHOPC Id‐protein. Two days later, animals were treated with Flt3‐L (10 µg per mouse/d, given i.p) for 10 consecutive days (d 0–9). On d 5 and d 11, myeloma‐specific immunoglobulin (IgHOPC) was administered s.c., together with incomplete Freund adjuvans (IFA) followed by the administration of IL‐2 (2 × 10·000/d given i.p) for 10 d (d 5–14). Whereas IgHOPC, Flt3‐L or IL‐2, given alone, did not elicit long‐term survival, the combination of IL‐2 or Flt3‐L with Id vaccination achieved a complete tumour rejection in 27% and 41% of mice respectively. However, the most powerful antimyeloma effects were induced by Flt3‐L + Id vaccination + IL‐2: 81% of the treated animals experienced long‐term survival (> 180 d). Both natural killer (NK) cells and CD8+ T cells may be involved in the antitumoral immune response. These data suggest that the combination of Flt3‐L and IL‐2 can be used to enhance the therapeutic effectiveness of clinical cancer vaccination protocols.
Belated diagnosis in three patients with rifampicin‐induced immune haemolytic anaemia
Authors: Ahrens Norbert , Genth Ramona , Salama Abdulgabar
Published: March 2003

Summary.  We report three patients who developed haemolysis following rifampicin treatment. Initially, autoimmune haemolytic anaemia (AIHA) of the warm type and/or an acute haemolytic transfusion reaction (AHTR) was suggested. The direct antiglobulin tests (DAT) were strongly positive for IgG and C3d, and tests for rifampicin‐dependent antibodies were positive in all three cases, featuring C‐specificity in one case. The outcome was fatal in two out of the three cases, presumably due to belated diagnosis. This shows that rifampicin may stimulate the production of autoantibodies (aab) and/or drug‐dependent antibodies (ddab), and that the resulting haemolytic syndrome bears similarities with AIHA and AHTR.
Translocations of 14q32 and deletions of 13q14 are common chromosomal abnormalities in systemic amyloidosis
Authors: Harrison Christine J. , Mazzullo Helen , Ross Fiona M. , Cheung Kan L. , Gerrard Gareth , Harewood Louise , Mehta Atul , Lachmann Helen J. , Hawkins Philip N. , Orchard Kim H.
Published: March 2003

Summary.  Systemic monoclonal immunoglobulin light chain amyloidosis (AL) is associated with clonal plasma cell dyscrasias that are often subtle and non‐proliferating. AL shares numerical chromosomal changes with multiple myeloma (MM) and monoclonal gammopathy of undetermined significance (MGUS). Illegitimate translocations involving the immunoglobulin heavy chain gene (IGH) at 14q32 and deletions of the long arm of chromosome 13, [del(13q)], commonly occur in MM, MGUS and plasma cell leukaemia. In AL IGH rearrangements have been identified but, to date, there are no reports of del(13q). In this study of 32 patients with AL, 24 with systemic and eight with localized disease, translocations involving IGH and del(13q) were found using dual‐colour interphase fluorescence in situ hybridization (FISH). IGH translocations were observed in 11 patients (37% overall and in 46% with systemic disease), of which nine had the IGH/CCND1 fusion from t(11;14)(q13;q32). Two showed IGH translocations other than the t(11;14) or t(4;14)(p16;q32). In one of these patients a breakpoint within the constant region of IGH between Cα1 and Cα2 was indicated. In the second a deletion covering Cα1 and Cα2 accompanied the translocation. Ten patients (27% overall and 33% of those with systemic disease) showed del(13q). The gain or loss of IGH and CCND1 signals provided evidence of numerical chromosomal changes in three patients.
Economics, health care systems and utilization of haematopoietic stem cell transplants in Europe
Authors: Gratwohl Alois , Passweg Jakob , Baldomero Helen , Horisberger Bruno , Urbano‐Ispizua Alvaro ,   for the Accreditation Committee of the European Group for Blood and Marrow Transplantation (EBMT)
Published: March 2003

Summary. Transplantation of haematopoietic stem cells from blood or bone marrow (HSCT) has seen rapid expansion. Increased costs and quality concerns present a challenge for health care providers. Information on factors influencing HSCT utilization is necessary. Data on 113 827 patients (37 761 allogeneic HSCT and 76 066 autologous HSCT), collected from 580 centres in 35 European countries between 1990 and 1999, were used. Economic factors, e.g. gross national product per capita, health care expenditure per capita and type of health care system were correlated with transplant rates (numbers of HSCT per 10 million inhabitants), team density (numbers of transplant teams per 10 million inhabitants) and increase in transplant numbers for each country. Annual numbers of HSCT increased in all European countries from 4234 in 1990 to 18 720 in 1999 irrespective of health care system. Economic strength and team density were the main determinants for transplant rate. This report reflects changes over the last decade and current status of HSCT in Europe. Economic strength, team density and hitherto unknown factors influence dissemination of the technology within Europe. These data provide a basis for health care planning, regulatory aspects and future research.
Thalidomide treatment in myelofibrosis with myeloid metaplasia
Authors: Elliott Michelle A. , Mesa Ruben A. , Li Chin‐Yang , Hook C. Christopher , Ansell Stephen M. , Levitt Ralph M. , Geyer Susan M. , Tefferi Ayalew
Published: March 2003

Summary.  Myelofibrosis with myeloid metaplasia (MMM) is uniquely characterized by macroscopic bone marrow stromal changes that are believed to be both reactive and cytokine mediated. Furthermore, a prognostically detrimental increase in bone marrow angiogenesis has recently been demonstrated. These observations suggest a potential therapeutic role for agents that are inhibitory to angiogenesis as well as cytokines that are pathogenetically implicated in MMM. In a prospective study of 15 patients with MMM, thalidomide treatment, starting at a dose of 200 mg/d, resulted in increased platelet counts (12 of 15 patients), increased haemoglobin level (3 of 15), a modest decrease in spleen size (3 of 12), increased bone marrow megakaryopoiesis (5 of 9) and decreased bone marrow angiogenesis (2 of 9). Undesirable haematological effects included pericardial extramedullary haematopoiesis in one patient, marked leucocytosis in two patients and extreme thrombocytosis in three patients. The thrombocytosis occurred in both patients with post‐thrombocythaemic myeloid metaplasia (PTMM) and was also associated with higher baseline levels of circulating CD34+ cells. Previously described toxicities of thalidomide were seen in the majority of patients and dose escalation to 400 mg/d was permitted in only two patients. In contrast, toxicity‐related dose reductions to 50 mg/d did not appear to lessen drug efficacy. We conclude that thalidomide has both beneficial and potentially adverse biological activity in MMM. A lower dose of the drug might be more tolerable without compromising therapeutic value. Patients with PTMM and/or markedly increased circulating CD34+ cell counts might be susceptible to thalidomide‐induced thrombocytosis.
Cell‐specific abnormal prenylation of Rab proteins in platelets and melanocytes of the gunmetal mouse
Authors: Zhang Qing , Zhen Lijie , Li Wei , Novak Edward K. , Collinson Lucy M. , Jang Elliott K. , J. Haslam Richard , Elliott Rosemary W. , Swank Richard T.
Published: March 2003

Summary. The mutant gunmetal mouse exhibits reduced rates of platelet synthesis, abnormalities of platelet alpha and dense granules and hypopigmentation. Several of these features resemble those of human alpha/delta platelet storage pool disease, grey platelet syndrome and Hermansky–Pudlak syndrome. Gunmetal mice have reduced levels of Rab geranylgeranyltransferase (RabGGTase), which adds lipophilic prenyl groups to the carboxyl terminus of Rab proteins. The degree of prenylation and the subcellular distribution of several Rab proteins were evaluated in mutant platelets, melanocytes and other tissues. Significant deficits in prenylation and membrane binding of most Rabs were observed in platelets and melanocytes. In contrast, minimal alterations in Rab prenylation were apparent in several other gunmetal tissues despite the fact that RabGGTase activity was equally diminished in these tissues. The mutant tissue‐specific effects are probably due to increased concentrations of Rab proteins in platelets and melanocytes. These experiments show that Rab proteins are differentially sensitive to levels of RabGGTase activity and that normal platelet synthesis, platelet organelle function and normal pigmentation are highly sensitive to the degree of prenylation and membrane association of Rab proteins. Further, the tissue‐specific effects of the gunmetal mutation suggest that RabGGTase is a potential target for therapy of thrombocytosis.
A novel cell line derived from de novo acute myeloblastic leukaemia with trilineage myelodysplasia which proliferates in response to a Notch ligand, Delta‐1 protein
Authors: Tohda Shuji , Sakano Seiji , Ohsawa Mai , Murakami Naomi , Nara Nobuo
Published: March 2003

Summary. A novel human leukaemia cell line, designated TMD7, was established from blast cells of a patient with de novo acute myeloblastic leukaemia with trilineage myelodysplasia (AML/TLD). As seen in the original blast cells, TMD7 cells expressed CD7, CD13, CD33 and CD34 and showed an abnormal karyotype containing −5, −7, −8, der(16)t(10;16)(q22;q13). The cells proliferated without added growth factors. Growth was stimulated with the addition of granulocyte colony‐stimulating factor (G‐CSF), granulocyte–macrophage CSF (GM‐CSF) and interleukin 3. Differentiation was not observed with the addition of various cytokines. As a cell line derived from AML/TLD has not been reported, TMD7 will be a useful tool as a model of AML/TLD cells. Recently, it was reported that the Notch system has crucial roles to regulate the self‐renewal and differentiation of haematopoietic stem cells. We found that TMD7 cells expressed Notch‐1 and Notch‐2 mRNA. The exposure to recombinant Delta‐1 protein, which was one of the Notch ligands, significantly stimulated the growth of TMD7 cells. This is the first human cell line which was shown to proliferate in response to Delta‐1, without artificially expressed Notch protein. Therefore, TMD7 will also be a useful tool to study the mechanism of the Notch–Notch ligand interaction.
The predictive value of clonogenic stem cell assays for the diagnosis of polycythaemia vera
Authors: Zwicky Caroline , Theiler Lorenz , Zbären Karin , Ischi Elisabeth , Tobler Anoreas
Published: March 2003

Summary.  Spontaneous growth of erythroid progenitor cells is one of the hallmarks of polycythaemia vera (PV). In vitro clonogenic assays can be used to support the diagnosis of PV. However, clonogenic assays are difficult to standardize, and their diagnostic value is accepted as a minor criteria for PV only in the case of positive results. Clonogenic assays need to be validated and normal ranges established carefully before they can be used for diagnostic purposes. In a retrospective study, we analysed the results of 146 consecutive clonogenic assays performed in patients with erythrocytosis between January 1992 and December 2000. Normal ranges were established on the basis of the results from 81 controls. The charts of the 146 patients were reviewed and the patients were allocated to the following diagnostic groups: PV (n = 38) and non‐primary erythrocytosis (NPE, n = 65) according to the criteria of Pearson & Messinezy. In total, 43 patients were excluded from the study because of missing data. The clonogenic assay results were correlated with the final clinical diagnosis of the patients. At a cut‐off value of 5 erythroid colony‐forming unit colonies per well, 29 out of 38 patients with PV showed positive results, and 9 out of 38 patients showed false‐negative results. In the NPE group, there were 64 out of 65 real‐negative results and 1 out of 65 false‐positive result. Thus the positive predictive value of the clonogenic assay for the diagnosis of PV was 97%. We conclude that clonogenic assays, in the case of positive results, and if carefully validated, are very useful and reliable tools for the diagnosis of PV.
T‐cell immune constitution after peripheral blood mononuclear cell transplantation in complete DiGeorge syndrome
Authors: Bensoussan Danièle , Le Deist Françoise , Latger‐Cannard Véronique , Grégoire Marie José , Avinens Odile , Feugier Pierre , Bourdon Violaine , André‐Botté Christine , Schmitt Claudine , Jonveaux Philippe , Eliaou Jean François , Stoltz Jean François , Bordigoni Pierre
Published: March 2003

Summary.  Complete DiGeorge syndrome (cDGS) is a congenital disorder characterized by typical facies, thymic aplasia, susceptibility to infections, hypoparathyroidism and conotruncal cardiac defect. Fetal thymus or post‐natal thymus tissue transplantations and human leucocyte antigen (HLA)‐genoidentical bone marrow transplantations were followed in a few cases by immune reconstitution. More recently, a peripheral blood mononuclear cell transplantation (PBMCT) was performed with an HLA‐genoidentical donor and followed by a partial T‐cell engraftment and immune reconstitution. We report a boy with cDGS, without cardiac defect, who suffered recurrent severe infections. At the age of 4 years, he underwent PBMCT from his HLA‐genoidentical sister. He received no conditioning regimen, but graft‐versus‐host disease (GVHD) prophylaxis was with oral cyclosporin A and mycophenolate mofetil. Toxicity was mild, with grade I acute GVHD. The patient is currently 2·5 years post‐PBMCT with excellent clinical performances. Mixed chimaerism can only be observed on the T‐cell population (50% donor T cells). T‐lymphocyte count fluctuated (CD3 more than 400 × 106/l at d 84 and CD4 more than 200 × 106/l at d 46). Exclusive memory phenotype T cells and absence of new thymic emigrants suggest expansion of infused T cells. T‐cell mitogen and tetanus antigen responses normalized a few months after transplantation. After immunizations, specific antibodies were produced. PBMCT from an HLA identical sibling could be an efficient treatment of immune deficiency in cDGS.
Marrow matrix metalloproteinases (MMPs) and tissue inhibitors of MMP in acute leukaemia: potential role of MMP‐9 as a surrogate marker to monitor leukaemic status in patients with acute myelogenous leukaemia
Authors: Lin Liang‐In , Lin Dong‐Tsamn , Chang Chi‐Jen , Lee Cheng‐Yeh , Tang Jih‐Luh , Tien Hwei‐Fang
Published: March 2003

Summary. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) were demonstrated to have important implications in the progression and invasiveness of many malignant disorders. In contrast, the biological significance of these molecules in human leukaemias is not clear. We determined the levels of MMP‐2, MMP‐9, TIMP‐1 and TIMP‐2 in the bone marrow of 37 patients with acute myelogenous leukaemia (AML) and 18 patients with acute lymphoblastic leukaemia (ALL) before chemotherapy. Nineteen bone marrow donors served as normal controls. After chemotherapy, sequential measurements were done during the course in 19 AML patients. The levels of TIMP‐1 and TIMP‐2 were significantly higher and MMP‐9 levels were significantly lower in the AML and ALL patients than in the normal controls. MMP‐2 levels were higher in ALL, but not AML patients, compared with controls. Moreover, the levels of marrow MMP‐2 and MMP‐9 did not parallel the numbers of leukaemic blasts in the peripheral blood. MMP‐9 levels were significantly lower in the AML patients who achieved a complete remission (CR) than in those who did not (8·71 ± 8·15 ng/ml vs 26·13 ± 27·75 ng/ml, P < 0·05). The AML patients with lower MMP‐9 levels (≤ 4·4 ng/ml) tended to have longer survival time than those with higher levels (> 12 months vs 4 months, P = 0·12). In addition, MMP‐9 levels in the AML patients at CR rose to the same range as the controls, but dropped again at relapse, demonstrating a close relationship of marrow MMP‐9 with disease status of AML. Therefore, we conclude that the level of marrow MMP‐9 may be a useful surrogate marker for monitoring disease status in AML and propose it as a potential prognostic factor.
Factors affecting long‐term outcome after allogeneic haematopoietic stem cell transplantation for acute myelogenous leukaemia: a retrospective study of 172 adult patients reported to the Austrian Stem Cell Transplantation Registry
Authors: Greinix Hildegard T. , Nachbaur David , Krieger Otto , Eibl Margit , Knöbl Paul , Kalhs Peter , Lutz Dieter , Linkesch Werner , Niederwieser Dietger , Hinterberger Wolfgang , Lechner Klaus , Rosenmayr Agathe , Gritsch Beate
Published: March 2003

Summary.Between 1982 and 2000, 172 patients with acute myelogenous leukaemia (AML) received haematopoietic stem cell transplants (SCT) from related (n= 132) or unrelated (n = 40) donors at four Austrian transplant centres and their results were reported to the Austrian Stem Cell Transplantation Registry. Conditioning for SCT consisted of cyclophosphamide and total body irradiation in 156 (91%) patients. Graft‐versus‐host disease (GVHD) prophylaxis was with standard cyclosporine and methotrexate in 95 (55%) patients. Median post‐transplant follow‐up was 5·6 years (range, 0·2‐‐16·7). Multivariate analysis of transplant‐related mortality (TRM) identified four variables associated with a lower risk: disease status of first complete remission (CR) at SCT, patient age of 45 years and younger, transplant performed during or after 1995, and lack of acute GVHD. Variables associated with significantly improved leukaemia‐free survival were: bone marrow as the stem cell source, disease status of first CR at SCT, and occurrence of chronic GVHD. In multivariate analysis, transplantation performed during or after 1995, first CR at SCT, occurrence of limited chronic GVHD and lack of acute GVHD grades III to IV were associated with increased overall survival. Based on these analyses, options for the improvement of results obtained with allogeneic SCT in patients with AML could be defined.
Phosphatidylinositol 3‐kinases are involved in the all‐trans retinoic acid‐induced upregulation of CD38 antigen on human haematopoietic cells
Authors: Lewandowski Daniel , Linassier Claude , Iochmann Sophie , Degenne Michel , Domenech Jorge , Colombat Philippe , Binet Christian , Hérault Olivier
Published: March 2003

Summary. All‐trans retinoic acid (ATRA) is a specific inducer of CD38 antigen on marrow CD34+ cells as well as on blast cells in acute promyelocytic and myeloblastic leukaemia. The CD38 antigen contributes to the control of blast cell proliferation, and the upregulation of CD38 might constitute an element in the pathogenesis of retinoic acid syndrome. The aim of this study was to determine whether phosphoinositide 3‐kinase (PI3‐K) is involved in the modification of CD38 antigen expression on myeloid cells, as PI3‐K plays a major role in the ATRA‐induced granulocytic differentiation of HL‐60 cells. We evaluated the effects of PI3‐K inhibitors (wortmannin and LY294002) on the levels of CD38 antigen and mRNA in HL‐60 and normal marrow CD34+ cells exposed to ATRA (1 µmol/l). The inhibitors prevented increase in CD38 mRNA expression and the overexpression of membrane CD38 antigen, without modification of the cytoplasmic level of this antigen. Interestingly, PI3‐K activity was also necessary for CD38 expression on normal marrow CD34+ cells and for the ATRA‐induced upregulation of CD157, a CD38‐related antigen. In conclusion, PI3‐K activity plays an essential role in the regulation of CD38 expression on human haematopoietic cells, and might constitute an interesting therapeutic target in haematological disorders involving CD38 overexpression.
Adenovirus transduction to effect CD40 signalling improves the immune stimulatory activity of myeloma cells
Authors: Bashey Asad , Cantwell Mark J. , Kipps Thomas J.
Published: March 2003

Summary. Neoplastic plasma cells from patients with myeloma fail to stimulate an effective anti‐myeloma immune response, which may be in part due to their deficient expression of immune accessory molecules. Attempting to alter this, we infected myeloma cell lines and patient‐derived primary myeloma cells with an adenovirus encoding CD154 (Ad‐CD154). Myeloma cells were made to express the CD154 transgene at multiplicity of infection (MOI) between 10 and 1000. Furthermore, infection of CD40positive myeloma cells with Ad‐CD154, but not an adenovirus encoding an irrelevant transgene, β‐galactosidase (Ad‐LacZ), induced enhanced expression of immune accessory molecules, such as CD54, HLA‐DR and CD70. In addition, Ad‐CD154‐infected myeloma cells could activate bystander CD40positive antigen‐presentingcells to express immune accessory molecules. Consequently, Ad‐CD154 infected myeloma cells stimulated proliferation in allogeneic mixed lymphocyte reactions (MLR). Finally, co‐infection of CD40negative myeloma cells with Ad‐CD154 and an adenovirus encoding CD40 (Ad‐CD40) induced expression of immune accessory molecules and enhanced the MLR stimulatory capacity of transduced myeloma cells. Collectively, these results indicate that infection of myeloma cells with Ad‐CD154 or Ad‐CD154/Ad‐CD40 can induce changes in myeloma cells that enhance their ability to induce cellular immune activation. As such, this approach may have potential application for immune therapy of patients with this disease.
DNA microarray analysis of T cell‐type lymphoproliferative disease of granular lymphocytes
Authors: Makishima Hideki , Ishida Fumihiro , Ito Toshiro , Kitano Kiyoshi , Ueno Shuichi , Ohmine Ken , Yamashita Yoshihiro , Ota Jun , Ota Masao , Yamauchi Kazuyoshi , Mano Hiroyuki
Published: March 2003

Summary. Lymphoproliferative disease of granular lympho‐ cytes (LDGL) is characterized by the clonal proliferationoflarge granular lymphocytes of either T‐ or natural killer cell origin. To better understand the nature of T cell‐type LDGL, we purified the CD4–CD8+ proliferative fractions from LDGL patients (n=4) and the surface marker‐matched T cells isolated from healthy volunteers (n=4), and compared the expression profiles of 3456 genes using DNA microarray. Through this analysis, we identified a total of six genes whose expression was active in the LDGL T cells, but silent in the normal ones. Interestingly, expression of the gene for interleukin (IL) 1β was specific to LDGL T cells, which was further confirmed by the examination of the serum level of IL‐1β protein. Given its important role in inflammatory reactions, the disease‐specific expression of IL‐1β may have a causative relationship with the LDGL‐ associated rheumatoidarthritis. Spectratyping analysis of the T‐cell receptor repertoire also proved the monoclonal or oligoclonal natureof LDGL cells. These data have shown that microarray analysis with a purified T‐cell subset is an efficient approach to investigate the pathological condition of Tcell‐type LDGL.
The role of common gamma chain in human monocytes in vivo; evaluation from the studies of X‐linked severe combined immunodeficiency (X‐SCID) carriers and X‐SCID patients who underwent cord blood stem cell transplantation
Authors: Ariga Tadashi , Yamaguchi Koji , Yoshida Jukei , Miyanoshita Akihiko , Watanabe Toshihide , Date Tadashi , Miura June–ichi , Kumaki Satoru , Ishii Naoto , Sakiyama Yukio
Published: March 2003

Summary. Expression of common gamma chain (γc) on monocytes was studied in five carriers of X‐linked severe combined immunodeficiency (X‐SCID) and two X‐SCID patients who underwent cord blood stem cell transplantation (CBSCT). Flow cytometric analysis revealed that both γc‐negative and positive monocytes co‐existed in X‐SCID carriers, whereas no γc‐negative T, B or NK cells were observed in them. Clonal analysis and reverse transcription polymerase chain reaction studies revealed that 13·2–45·0% of monocytes from these carriers expressed the mutant γc message. X‐SCID patients who received CBSCT persistently possessed the majority of γc‐negative monocytes with a good clinical course. These results, together, may indicate that γc is not essential for monocyte development/function in vivo.
High‐dose cytarabine in upfront therapy for adult patients with acute lymphoblastic leukaemia
Authors: Hallböök Helene , Simonsson Bengt , Ahlgren Thomas , Björkholm Magnus , Carneskog Jan , Grimfors Gunnar , Hast Robert , Karlsson Karin , Kimby Eva , Lerner Richard , Linder Olle , Linderholm Mats , Löfvenberg Eva , Malm Claes , Nilsson Per‐Gunnar , Paul Christer , Stenke Leif , Stockelberg Dick , Tidefelt Ulf , Turesson Ingemar , Uden‐Blome Ann‐Marie , Vilen Lars , Wahlin Anders , Winquist Ingemar , Smedmyr Bengt
Published: March 2003

Summary. In this national study, we have evaluated a new intensive chemotherapy protocol for adult patients with untreated acute lymphoblastic leukaemia (ALL). One hundred and fifty‐three patients with median age 42 years received induction therapy with high‐dose cytarabine (Ara‐C), cyclophosphamide, daunorubicin, vincristine and betamethasone. A high complete remission (CR) rate (90%) was achieved in patients < 60 years compared with 70% in patients > 60 years (P = 0·004). The estimated 3 year overall survival for all patients was 29% (CI 21–36%) and the estimated continuous complete remission (CCR) at 3 years for the patients achieving CR according to the protocol was 36% (CI 27–45%). A favourable pretreatment characteristic was pre‐B phenotype, especially for patients < 40 years without any high‐risk factor, with an estimated CCR at 3 years of 62% (CI 41–82%). Stem cell transplantation (SCT) as post‐remission therapy, mainly for high‐risk patients, gave an estimated 3 year disease free survival (DFS) after SCT of 39% (CI 24–54%). No significant differences in DFS could be found between autologous, related or unrelated donor transplantation. We conclude that this intensive protocol resulted in a high CR rate combined with acceptable side‐effects and a favourable CCR for patients with pre‐B ALL.
Sex ratios and the risks of haematological malignancies
Authors: Cartwright Ray A. , Gurney Karen A. , Moorman Anthony V.
Published: March 2003

Summary. Although the sex of an individual confers one of the greatest of the known risks for contracting leukaemia and lymphomas, very little attention is paid to these risks. It is the purpose of this paper to stimulate further research in this area. The sex rate ratios are presented for the commoner haematological malignancies. The male excess in the lymphoid cancers is most marked in the youngest age group in non‐Hodgkin's lymphoma and Hodgkin's disease, while acute lymphoblastic leukaemia shows equal sex ratios in the childhood peak. Both chronic lymphocytic leukaemia and lymphocytic lymphoma display an unusual pattern, hitherto undescribed, with a large male excess specific to the 40s and 60s age groups. The myeloid sex ratios are all characterized by slight female excess in early adulthood followed by marked male excess. The reasons for these patterns are discussed.
Effect of anti‐CD20 (rituximab) on resistant thrombocytopenia in autoimmune lymphoproliferative syndrome
Authors: Heelan Bridget T. , Tormey Vincent , Amlot Peter , Payne Elspeth , Mehta Atul , Webster A. David B.
Published: March 2003

Summary.  Fas (CD95) plays an important role in apoptosis. Patients with defects in Fas have an autoimmune lymphoproliferative syndrome (ALPS) characterized by lymphadenopathy, autoimmune cytopenias and an increased incidence of lymphomas. There are approximately 70 known cases described worldwide. The autoimmune cytopenias are difficult to treat in this group. We describe a patient with a defect in the death domain of the FAS molecule who had autoimmune thrombocytopenia resistant to conventional therapy but which responded to a combination of rituximab and vincristine.
A Ser752→Pro substitution in the cytoplasmic domain of β3 in a Glanzmann thrombasthenia variant fails to prevent interactions between the αIIbβ3 integrin and the platelet granule pool of fibrinogen
Authors: Nurden Paquita , Poujol Christel , Winckler Joelle , Combrié Robert , Caen Jacques P. , Nurden Alan T.
Published: March 2003

Summary. A Glanzmann thrombasthenia variant with a β3 Ser752→Pro cytoplasmic domain substitution has platelets that fail to aggregate or bind soluble fibrinogen (Fg) after activation. Despite this, Fg is normally present in the α‐granules. We have used immunoelectron microscopy to examine the reactivity of Fg with the different pools of αIIbβ3 in the patient's platelets. Immunogold labelling was performed on cryosections using an anti‐ligand‐induced binding site (LIBS) monoclonal antibody (mAb), which binds to αIIbβ3 only when Fg is bound, or a mixture of two anti‐receptor‐induced binding site (RIBS) mAbs that specifically recognize receptor‐bound Fg. Labelling of the α‐granule membrane and channels of the surface‐connected canalicular system in unstimulated platelets confirmed that the mutated αIIbβ3 retains the capacity to transport Fg. When the patient's platelets were stimulated with ADP in the presence of Fg, as expected there was a much‐decreased activation of surface‐exposed αIIbβ3. However, thrombin‐induced activation was associated with both secretion and a rapid increase in the labelling of internal membrane systems by anti‐RIBS and anti‐LIBS mAbs, with mobilization of theinternal Fg pool. Yet labelling on the surface of the patient's platelets was transient. Our studies implied that αIIbβ3 in platelets may bind fibrinogen in different activation states and that this patient specifically lacked high‐affinity binding.
Increased angiogenesis in bone marrow of children with acute lymphoblastic leukaemia has no prognostic significance
Authors: Pulè Martin A. , Gullmann Christian , Dennis Deirdre , McMahon Corrina , Jeffers Michael , Smith Owen P.
Published: March 2003

Summary. The importance of angiogenesis for the growth and viability of solid tumours has been established. Similarly, prognostic information may be gained from the extent of angiogenesis in these tumours. Haematopoietic malignancies should have equal requirements for angiogenesis and important prognostic information may be derived from quantification of bone marrow angiogenic activity. We retrospectively investigated 82 bone marrow trephine biopsies from 41 children with acute lymphoblastic leukaemia (ALL) at diagnosis and following treatment. Nine normal bone marrow trephines from age‐matched children were also analysed as controls. The microvessels were stained immunohistochemically with anti‐Factor VIII‐related antigen (antivWF) and antithrombomodulin (anti‐THR). Angiogenesis was quantified manually by two independent observers and was highly reproducible (Pearson's r = 0·91). Staining with anti‐vWF and anti‐THR was highly specific for microvessels and thetwo stains closely correlated (r = 0·68). Microvessel densities (MVD) at presentation were significantly increased in the majority of patients in comparison with controls (P < 0·0001) and MVD dropped towards normal in remission (P < 0·0001). Of interest, the difference in total vessel counts between leukaemic and normal/remission marrows was contributed solely by small microvessels. There was no significant difference in MVD at presentation or remission from children in poor prognostic groups or those who subsequently relapsed. Similarly, we could not find an association with age, sex, cytogenetic abnormality or disease phenotype.
Intensive chemotherapy in patients with chronic myelogenous leukaemia (CML) in accelerated or blastic phase – a report from the Swedish CML Group
Authors: Axdorph Ulla , Stenke Leif , Grimfors Gunnar , Carneskog Jan , Hansen Jan , Linder Olle , Ljungman Per , Löfvenberg Eva , Malm Claes , Simonsson Bengt , Turesson Ingemar , Vilén Lars , Udén Anne‐Marie , Björkholm Magnus ,   for the Swedish CML Group
Published: March 2003

Summary. In attempting to restore the chronic phase (CP) of chronic myelogenous leukaemia (CML), the Swedish CML group utilized an intensive chemotherapy protocol for 83 patients (aged 16–79 years) in accelerated (AP, n = 22) or blastic phase (BC, n = 61). Most patients received a combination of mitoxantrone (12 mg/m2/d) and etoposide (100 mg/m2/d) together with cytosine arabinoside (1 g/m2 b.i.d) for 4 d. Overall, 39 patients (47%) achieved a second CP (CP2)/partial remission (PR). Responding patients < 65 years were eligible for ablative chemotherapy followed by an allogeneic (SCT) or a double autologous stem cell transplant (ASCT). Seventeen of 34 responders < 65 years failed to proceed to transplantation as a result of early disease progression (n = 15) or disease‐related complications (n = 2). The remaining 17 patients underwent SCT (n = 9; including four unrelated donor SCT) or ASCT (n = 8). Only one of the eight ASCT patients had a second ASCT; the remaining seven failed because of progression (n = 5) or hypoplasia (n = 2). The median duration of CP2/PR was 6 months (range 1–72 months). Five patients achieved a longer CP2/PR than CP1. The 1 year survival was 70% for SCT/ASCT patients (median survival 21 months), 50% for responding patients overall, but only 7% for non‐responders (P < 0·001). Three SCT/ASCT patients are long‐term survivors (65+, 66+ and 73+ months). In conclusion, approximately half of the patients achieved a CP2/PR after intensive chemotherapy, with a clear survival advantage for responders vs non‐responders. Subsequent SCT/ASCT was feasible for half of the responders (< 65 years), and one individual underwent double ASCT. Novel therapeutic options for CML patients in AP/BP are needed.
Treatment of high‐risk acute myelogenous leukaemia by myeloablative chemoradiotherapy followed by co‐infusion of T cell‐depleted haematopoietic stem cells and culture‐expanded marrow mesenchymal stem cells from a related donor with one fully mismatched human leucocyte antigen haplotype
Authors: Lee Seung Tae , Jang Joon Ho , Cheong June‐Won , Kim Jin Seok , Maemg Ho‐Young , Hahn Jee Sook , Ko Yun Woong , Min Yoo Hong
Published: March 2003

Summary.  A 20‐year‐old woman with high‐risk acute myelogenous leukaemia was transplanted with granulocyte colony stimulating factor (G‐CSF)‐mobilized peripheral blood CD34+ haematopoietic stem cells and bone‐marrow‐derived mesenchymal stem cells (MSC) from her human leucocyte antigen haplotype‐mismatched father after myeloablative conditioning therapy. The patient engrafted rapidly and had no acute or chronic graft‐versus‐host disease. Since transplantation, the patient has shown an enduring trilineage haematological complete response without any evidence of leukaemia relapse at 31 months. We suggest that MSC can be used effectively for genetically haploidentical haematopoietic stem cell transplantation for acute leukaemia.
Tumoral non‐amyloidotic monoclonal immunoglobulin light chain deposits (‘aggregoma’): presenting feature of B‐cell dyscrasia in three cases with immunohistochemical and biochemical analyses
Authors: Rostagno Agueda , Frizzera Glauco , Ylagan Lourdes , Kumar Asok , Ghiso Jorge , Gallo Gloria
Published: March 2003

Summary. Tumoral monoclonal immunoglobulin (Ig) light chain non‐fibrillar deposits (‘aggregomas’), which can be considered analogous to solitary light chain amyloidomas, are a rare presenting feature of B‐cell dyscrasias. It is not certain if they are truly localized or if in reality they represent an initial expression of a silent systemic non‐amyloid light chain deposition disease (LCDD). This report describes three patients, two of whom presented with cervical masses and the third with a solitary lung nodule, each comprising granular aggregates of monoclonal kappa light chain. Extracted deposits from the lymph node of one patient were shown by N‐terminal amino acid sequence analysis to belong to the variable‐region kappa I (Vκ I) light chain subgroup, the first reported κ‐LCDD protein encoded by the L9 gene and the first report of an expressed protein related to this gene. Extracted deposits from the lung nodule of the second patient belonged to the Vκ IV light chain subgroup encoded by the B3 germ line gene. The N‐terminal amino acid sequences of the light chains from theaggregomaswere compared with the related germ line sequences and to the N‐terminal amino acid sequences of the nine other known κ‐LCDD light chains reported thus far from patients with systemic LCDD.
Prognosis of myelodysplasic patients: non‐parametric multiple regression analysis of populations stratified by mean corpuscular volume and marrow myeloblast number
Authors: Tennant Graham B. , Al‐Sabah Ala I , Burnett Alan K.
Published: March 2003

Summary. Myelodysplastic (MDS) patients at diagnosis (n = 162) were analysed by the International Prognostic Scoring System (IPSS). The two intermediate groups were not significantly different. The IPSS was of limited value in predicting survival of MDS patients after preliminary separation into subgroups with < 5%, or ≥ 5%, myeloblasts. Cox's proportional hazards analysis of these subgroups enabled discrimination of highly significant prognostic groups. In both subgroups, longer survival was associated with macrocytosis. Mean corpuscular volume (MCV) and marrow myeloblast number were used to define four groups with prognostic significance similar to the IPSS. A low‐risk group was described by macrocytosis associated with < 5% myeloblasts and high risk was related to blast counts ≥ 5% and MCV < 100 fl. Further analysis defined factors identifying very high‐risk patients and those with benign disease, together with many intermediate survival patterns. Results were consistent in two time‐separated patient groups.