bluuebunny/crossref_metadata_2025 huggingface dataset:
p53 allele deletion and protein accumulation occurs in the absence of p53 gene mutation in T‐prolymphocytic leukaemia and Sezary syndrome
Authors: Brito‐Babapulle V. , Hamoudi R. , Matutes E. , Watson S. , Kaczmarek P. , Maljaie H. , Catovsky D.
Published: March 2003

In a series of 24 patients with chronic T‐lymphoid disorders [13 T‐prolymphocytic leukaemia (T‐PLL) and 11 Sezary syndrome] we have studied (i) chromosome 17p abnormalities and p53 allele deletion by fluorescence in situ hybridization; (ii) mutation in the exons of the p53 gene by direct DNA sequencing; and (iii) p53 protein expression by immunocytochemistry and, in some cases, also by flow cytometry with DO‐1, a monoclonal antibody to the p53 protein. The study revealed p53 deletion and accumulation of p53 protein in the absence of mutation in the exons that included the hot‐spots and differs from that described in B‐prolymphocytic leukaemia. Seven T‐PLL and five Sezary syndrome patients had p53 overexpression, and five T‐PLL and nine Sezary syndrome patients showed p53 deletion. Although the majority of cases with p53 accumulation had p53 deletion, the proportion of cells with the deletion did not correlate with the proportion of cells positive for p53 expression. Two cases of T‐PLL showed strong p53 expression in the absence of p53 deletion, and one case of Sezary syndrome with p53 deletion in 97% of cells did not express p53. These findings suggest that a non‐mutational mechanism exists for the accumulation of p53 protein in these T‐cell disorders. The oncogenic effect of the accumulating wild‐type protein has been reported in other malignancies. Whether haploidy resulting from p53 deletion contributes to this mechanism has yet to be determined. Alternatively, the frequent loss of the p53 gene could be associated with the deletion of an adjacent gene, which could be involved in the pathogenesis of these diseases.
Withdrawal of enzyme replacement therapy in Gaucher's disease
Authors: Elstein Deborah , Abrahamov Ayala , Hadas‐Halpern Irith , Zimran Ari
Published: March 2003

Although enzyme replacement therapy is safe and effective in ameliorating the signs and symptoms of Gaucher's disease, some patients have withdrawn from treatment. The purpose of this study was to evaluate the response to withdrawal and to discuss the implications for patients currently on unaltered therapy regimens since the advent of treatment. Fifteen patients, who had been treated with enzyme replacement for 5–56 months and then withdrew for 8–47 months, were assessed for changes in haematological parameters and in liver and spleen index volume. Despite non‐uniformity of duration of on and off periods, degree of organomegaly, anaemia and thrombocytopenia, most patients did not revert to respective baseline values in most parameters after withdrawal. None of the patients suffered exacerbation of bone involvement or had new or aggravated pulmonary hypertension. Adult patients with stable Gaucher's disease may be withdrawn from therapy for circumscribed periods without forfeiting most gains accrued during enzyme therapy. Therefore, stopping and restarting may be considered in some patients. Alternatively, maintenance at reduced dosage and/or frequency may be appropriate in some adult patients who are stable or non‐responsive after the first years of enzyme therapy. This caveat does not apply to children.
In vitro anti‐tumour activity of anti‐CD80 and anti‐CD86 immunotoxins containing type 1 ribosome‐inactivating proteins
Authors: Bolognesi Andrea , Polito Letizia , Tazzari Pier Luigi , Lemoli Roberto M. , Lubelli Chiara , Fogli Miriam , Boon Louis , De Boer Mark , Stirpe Fiorenzo
Published: March 2003

Immunotoxins specific for the CD80 and CD86 antigens were prepared by linking three type 1 ribosome‐inactivating proteins (RIPs), namely bouganin, gelonin and saporin‐S6, to the monoclonal antibodies M24 (anti‐CD80) and 1G10 (anti‐CD86). These immunotoxins showed a specific cytotoxicity for the CD80/CD86‐expressing cell lines Raji and L428. The immunotoxins inhibited protein synthesis by target cells with IC50s (concentration causing 50% inhibition) ranging from 0·25 to 192 pmol/l as RIPs. The anti‐CD80 immunotoxins appeared 1–2 log more toxic for target cells than the anti‐CD86 ones. Immunotoxins containing saporin and bouganin induced apoptosis of target cells. The toxicity for bone marrow haemopoietic progenitors of these conjugates was also evaluated. Bouganin and related immunotoxins at concentrations up to 100 nmol/l did not significantly affect the recovery of committed progenitors or of more primitive cells. The saporin‐containing immunotoxins at concentrations ≥ 1 nmol/l showed some toxicity on colony‐forming unit cells (CFU‐C). The expression of the CD80 and CD86 molecules is prevalently restricted to antigen‐presenting cells and is also strong on Hodgkin and Reed–Sternberg cells in Hodgkin's disease. Present results suggest that immunotoxins targeting type 1 ribosome‐inactivating proteins to these antigens could be considered and further studied for the therapy of Hodgkin's disease or other CD80/CD86‐expressing tumours.
Platelet ultrastructural abnormalities in three patients with type 2B von Willebrand disease
Authors: Nurden Paquita , Chretien Fabrice , Poujol Christel , Winckler Joelle , Borel‐Derlon Annie , Nurden Alan
Published: March 2003

Several reports have described the presence of giant platelets in patients with type 2B von Willebrand disease (VWD). We have now characterized the ultrastructural changes in platelets from three unrelated patients with type 2B VWD and different mutations within exon 28 of the von Willebrand factor (VWF) gene. Electron microscopy showed that each of these subjects had an increased proportion of large platelets when compared with those of a patient with type 2A VWD or control subjects. Immunogold labelling for VWF was performed. Large masses detected by anti‐VWF antibody were seen not only on the platelet surface, but also inside the platelet surface‐connected canalicular system (SCCS) when ultrathin sections were labelled. This suggested translocation of the abnormally bound VWF from the platelet surface. Labelling of the α‐granules was eccentric as for normal platelets. Labelling for glycoprotein (GP) Ib was seen on the surface and within the SCCS, suggesting co‐localization with the bound VWF. However, there was no evidence for VWF in endosomes or other endocytic vesicles. The presence of platelet‐bound VWF was not accompanied by high levels of platelet activation, as detected by electron microscopy, or by using monoclonal antibodies against P‐selectin or activation‐dependent determinants on GP IIb‐IIIa in flow cytometry. Intriguingly, platelet ultrastructure often resembled that seen in patients with congenital thrombocytopathies characteristic of giant platelet syndromes.
Contrasting effects of HSP72 expression on apoptosis in human umbilical vein endothelial cells and an angiogenic cell line, ECV304
Authors: Lucas M. , Rose P. E. , Morris A. G.
Published: March 2003

The effect of overexpression of heat shock protein (HSP)72 on apoptosis induced by different stimuli in human umbilical vein endothelial cells (HUVECs) and the angiogenic cell line, ECV304, was studied. Transient overexpression of HSP72 was achieved using an adenoviral vector (Advhsp72) and apoptosis was induced by heat shock, tumour necrosis factor (TNF)‐α with cycloheximide (CHX), lipopolysaccharide (LPS) with TNF‐α and verocytotoxin (VT). Apoptosis induced by heat shock was reduced by HSP72 expression. However, HSP72 expression in HUVECs increased apoptosis induced by TNF‐α/CHX, LPS and VT measured by flow cytometric analysis of propidium iodide (PI)‐stained permeabilized cells. In contrast, apoptosis in ECV304 induced by the same stimuli was reduced by HSP72 expression. No difference was seen in cells transduced with a control adenoviral vector expressing β‐galactosidase. These data imply that induction of HSP72 in cells modulates responses to apoptotic stimuli, but that the nature of the response varies with the cell type. However, it is clear that in situations where apoptosis may be part of a pathological process, HSP72 induction, for example by reperfusion injury, may exacerbate the process.
Cytogenetic and molecular diagnosis of chromosome 5 deletions in myelodysplasia
Authors: Westbrook C. A. , Hsu W.‐T. , Chyna B. , Litvak D. , Raza A. , Horrigan S. K.
Published: March 2003

Deletions of chromosome 5, del(5q), are frequently observed in myelodysplasia (MDS). We evaluated molecular detection of loss of heterozygosity (LOH) as a diagnostic method to detect del(5q) in a series of 60 MDS cases at a single institution. LOH was compared to cytogenetics on the same clinical specimen, resolving ambiguous cases by fluorescent in situ hybridization (FISH) and additional LOH. There was poor concordance between molecular and cytogenetic results, but most discrepancies could be resolved by FISH and additional LOH. Molecular analysis was of low sensitivity because most cases contained a relatively high proportion of cells without del(5q), but it was accurate, while cytogenetics overestimated the proportion of cells with del(5q) and failed to detect some cases with complex rearrangements. Minor clones were detected both by FISH and LOH. Overall, we found an incidence of 23% (14 of 60 cases) for del(5q) in MDS. The results also suggest that there is a high degree of genetic heterogeneity in the cellular population of MDS. Although del(5q) is common in MDS, it may not be present in all cells, leading to diagnostic challenges.
Hepatic iron concentration combined with long‐term monitoring of serum ferritin to predict complications of iron overload in thalassaemia major
Authors: Telfer P. T. , Prestcott E. , Holden S. , Walker M. , Hoffbrand A. V. , Wonke B.
Published: March 2003

Clinical complications of transfusional iron overload are still common in patients with thalassaemia major (TM) and it is not clear how best to monitor body iron stores during long‐term follow‐up to anticipate tissue damage. In this study, we have reviewed a group of 32 patients who underwent liver biopsy between 1984 and 1986. We developed a method of assessing the trend in serum ferritin (TSF) during long‐term monitoring and compared this with mean serum ferritin (MSF) and initial liver iron (LI) concentration to determine whether, individually or in combination, they were accurate in predicting clinical outcome. LI levels were low (< 7 mg/g), medium (7–15 mg/g) and high (> 15 mg/g dry weight) in 15, 7 and 10 patients respectively. MSF was low (< 1500 μg/l), medium (1500–2500 μg/l) and high (> 2500 μg/l) in 10, 14 and 8 patients. TSF was low, medium and high risk in 9, 9 and 11 out of 29 evaluable patients. During a median follow‐up of 13·6 years (range 2·3–14·8 years) after biopsy, nine patients died and an additional three patients developed heart failure. Hypothyroidism developed in five, hypoparathyroidism in four, and diabetes mellitus in seven patients. Cirrhosis developed in four of 10 evaluable patients. The clinical end‐point of death or cardiac failure was significantly associated with increasing iron load using all three means of assessment. Although numbers were insufficient for statistical analysis, MSF or TSF were more closely associated with complications of iron overload than LI. There was no clear additional value in combining LI with MSF or TSF. The data show that quantitation of liver iron from a single liver biopsy has little value in long‐term monitoring of iron stores. Most complications can be avoided if ferritin levels can be brought down to <1500 μg/l.
Hepatocyte growth factor is secreted by osteoblasts and cooperatively permits the survival of haematopoietic progenitors
Authors: Taichman Russell S. , Reilly Marcelle J. , Verma Rama S. , Ehrenman Karen , Emerson Stephen G.
Published: March 2003

Human osteoblasts (HOBs) support the growth of human haematopoietic progenitor cells, and support the survival and limited expansion of long‐term culture‐initiating cells. Using human CD34+ cells and the murine myelomonocytic cell line NFS‐60 as targets, we previously found that one component of HOB‐derived haematopoietic activity is cell‐associated granulocyte colony‐stimulating factor (G‐CSF). However, antibody failed to neutralize all the activity, suggesting that more than one factor supports haematopoietic cells. In the present investigations, we asked whether the HOB‐derived, non‐G‐CSF secreted activity was as a result of other known growth factors. We found that, among the cytokines expressed by HOBs, only hepatocyte growth factor (HGF) and G‐CSF stimulated NFS‐60 cell proliferation. HOB cells and osteosarcoma cells secreted biologically active HGF, although the levels varied considerably. Moreover, addition of neutralizing HGF antibody to CD34+ cell/HOB co‐cultures resulted in a significant reduction (≈50%) in the ability of the HOBs to support haematopoietic progenitor cells. These results suggest that a major component of osteoblast‐derived haematopoietic activity is HGF. Secretion of HGF, in concert with cell‐associated cytokines such as G‐CSF, may account for the stem cell‐stimulating activities of osteogenic cells and, thereby, the unique stem cell‐supporting role of the osteoblasts within the bone marrow microenvironment.
Fludarabine‐based stem cell transplantation protocol for Fanconi's anaemia in myelodysplastic transformation
Authors: McCloy M. , Almeida A. , Daly P. , Vulliamy T. , Roberts I. A. G. , Dokal I.
Published: March 2003

Allogeneic stem cell transplantation (SCT) represents the treatment of choice for severe bone marrow (BM) failure in patients with Fanconi's anaemia (FA). However, for FA patients developing leukaemic or myelodysplastic transformation, the results of SCT are much less encouraging. We present a 17‐year‐old girl with myelodysplastic transformation of FA (refractory anaemia with excess blasts) and oculocutaneous albinism, who was treated by sibling SCT using conditioning with fludarabine, cyclophosphamide (CY) and anti‐lymphocyte globulin (ALG). She had rapid engraftment with no toxicity and no graft‐versus‐host disease (GVHD). Twenty‐two months after SCT, she had 100% donor chimaerism on Southern blot analysis.
The granulocyte colony‐stimulating factor receptor supports erythroid differentiation in the absence of the erythropoietin receptor or Stat5
Authors: Millot Gaël A. , Svinarchuk Fédor , Lacout Catherine , Vainchenker William , Dumenil Dominique
Published: March 2003

To evaluate the functional conservation of signal transduction mechanisms between haematopoietic receptors and to characterize the molecules activated in this phenomenon, we introduced granulocyte colony‐stimulating factor receptor (G‐CSFR) cDNA into mouse fetal liver cells using a retroviral vector. In semi‐solid medium assays, G‐CSFR‐infected cells gave rise to all types of colonies [granulocyte‐macrophage (GM), megakaryocyte (MK) and mixed lineage (GEMM) colony‐forming units (CFU) and erythroid burst‐forming units (BFU‐E)] in the presence of G‐CSF alone. The direct effect of G‐CSF on erythroid differentiation of G‐CSFR‐transduced erythroid progenitors was demonstrated by the development of erythroid colonies using G‐CSFR‐expressing Lin− cells cloned at one cell per well in liquid culture in the presence of G‐CSF. Interestingly, while Stat5, but not Stat3, was activated in erythroid cells in response to erythropoietin (EPO), both were activated in erythroid and granulocytic cells stimulated by G‐CSF. Furthermore, G‐CSF induced the growth of erythroid colonies from G‐CSFR‐expressing fetal liver cells from EPO receptor−/− (EPO‐R−/−) or Stat5a−/− Stat5b−/− mice, demonstrating that erythroid differentiation can occur in the absence of EPO‐R or Stat5. These data show that forced expression of G‐CSFR allows G‐CSF‐dependent multilineage proliferation and differentiation of haematopoietic progenitors and rescues EPO‐R−/− erythroid cells. While G‐CSF induces Stat5 activation in G‐CSFR‐expressing erythroid cells, this activation is not necessary for the terminal erythroid differentiation induced by G‐CSF.
Changes in the pattern of TCR Vβ repertoire expression after bone marrow transplant is linked to the HLA haplotype in humans
Authors: Lonial Sagar , Bomberger Catherine , Waller Edmund K.
Published: March 2003

Neutrophil, monocyte, natural killer‐ and B‐cell number and function are rapidly restored after bone marrow transplant (BMT), whereas T‐cell reconstitution is often quite delayed. Our hypothesis was that Vβ T‐cell receptor (TCR) repertoire diversity among recipients of allogeneic BMT is influenced by the expression of major and minor HLA antigens in the host. The study population comprised unmanipulated and CD34+‐selected allogeneic bone marrow grafts, autologous peripheral blood stem cell transplants and recipients of volunteer unrelated donor (VUD) bone marrow transplants. Using flow cytometry, the relative frequencies of 18 Vβ TCR families were determined and ranked for each time point studied. Comparisons and correlations were made between paired blood samples obtained within a single patient over time, and between donors and their recipients. The pattern of the Vβ TCR repertoire from allogeneic recipients and their HLA‐matched donors was very similar, with a correlation coefficient (CC) of 0·59. This similarity was not as marked in VUD pairs (CC = 0·32). By 3 months after transplant, the pattern of the Vβ TCR repertoire in recipients of HLA‐matched sibling transplants was more similar to the pattern seen in pretransplant recipients than to the donor pattern (CC = 0·40 vs. 0·31). Our data suggest that both major and minor HLA antigens influence Vβ TCR repertoire diversity and reconstitution after BMT.
Human herpesvirus 8 DNA sequences are present in bone marrow from HIV‐negative patients with lymphoproliferative disorders and from healthy donors
Authors: Azzi A. , Fanci R. , De Santis R. , Ciappi S. , Paci C.
Published: March 2003

Bone marrow (BM) from patients affected by multiple myeloma (MM), exhibiting monoclonal gammopathy of undetermined significance (MGUS) or with non‐Hodgkin lymphoma (NHL) as well as from healthy donors were investigated for the presence of human herpesvirus‐8 (HHV‐8) DNA sequences. ORF 26 sequences were detected in 36–56% of the patients and in 29% of the controls. In a few cases, two other HHV‐8 DNA sequences were also detected. These observations indicate that the presence of the HHV‐8 genome in BM is relatively common in different groups of patients as well as in healthy individuals and do not support an alleged role for HHV‐8 in MM.
Provision of platelet support for fetuses and neonates affected by severe fetomaternal alloimmune thrombocytopenia
Authors: Ranasinghe E. , Walton J. D. , Hurd C. M. , Saul L. , Smith G. , Campbell K. , Ouwehand W. H.
Published: March 2003

Severe fetomaternal alloimmune thrombocytopenia requires urgent treatment with compatible platelet concentrates. As prompt treatment is sometimes delayed owing to the unavailability of compatible platelets, we established an accredited platelet donor panel to provide effective and timely transfusion support for fetal and neonatal therapy. After a mass screening programme of over 60 000 blood donations, 45 HPA‐1a‐negative donors with no antibodies to HPA, HLA, red cell antigens and granulocytes/lymphocytes, and with low titre anti‐A and/or ‐B were accredited. All accredited donors were fully genotyped for HPA‐1, ‐2, ‐3 and ‐5 by PCR‐SSP. Ninety‐one per cent of the accredited donors were also negative for HPA‐5b.
Killer T‐cell induction in patients with blastic natural killer cell lymphoma/leukaemia: implications for successful treatment and possible therapeutic strategies
Authors: Yamada Osamu , Ichikawa Masako , Okamoto Takamitsu , Park Chisen , Motoji Toshiko , Mizoguchi Hideaki , Shibuya Akira
Published: March 2003

A rare form of putative natural killer (NK) cell lymphoma called blastic NK cell lymphoma appears to be clinicopathologically distinctive in showing a homogenous lymphoblast, variable expression of CD2, CD4, CD56 and TdT, negative for surface CD3, T‐cell receptor antigen, CD16, CD34 and lack of association with Epstein–Barr virus (EBV). We report two patients with blastic NK cell lymphoma and describe the interesting clinical studies. The patients presented with cutaneous plaques. Both patients had adenopathy, and one had marrow involvement at presentation. Unlike in many NK and NK‐like T‐cell disorders, azurophilic cytoplasmic granules were absent. They expressed intermediate density CD45. In addition, the cells were positive for HLA‐DR, CD2, CD4, CD56 and TdT, and negative for EBV transcripts. In spite of the advanced clinical stage, complete remission was achieved by conventional chemotherapy. After interleukin 2 expansion of tumour‐infiltrating bone marrow and lymph node cells from the patients, cytotoxic T‐cell lines with rearranged T‐cell receptor genes were established. They showed specific killing activity against autologous tumour cells in an MHC‐restricted fashion, with possible implications for treatment. In addition, upon cessation of maintenance chemotherapy, one patient developed overt leukaemia with blasts expressing CD33 antigens, suggesting a continuous spectrum of blastic NK cell lymphoma to myeloid/NK cell precursor acute leukaemia.
Interferon α and zidovudine therapy in adult T‐cell leukaemia lymphoma: response and outcome in 15 patients
Authors: Matutes E. , Taylor G. P. , Cavenagh J. , Pagliuca A. , Bareford D. , Domingo A. , Hamblin M. , Kelsey S. , Mir N. , Reilly J. T.
Published: March 2003

Adult T‐cell leukaemia lymphoma (ATLL) is an aggressive disease caused by the human T‐lymphotropic virus 1 (HTLV‐I) with a short survival. Responses to interferon alpha (IFN‐α) and zidovudine (AZT) have been documented but not with long‐term follow‐up. We treated 15 ATLL patients with IFN and AZT. Eleven patients had acute ATLL, two had lymphoma and two smouldering ATLL, with progression. The main features were: organomegaly (14), skin lesions (10), high white blood cell (WBC) count (11) and hypercalcaemia (9). Eleven patients had previously received chemotherapy and one had received an autograft. At the time of the study, seven patients had progressive disease and eight were in partial or complete clinical remission. Responses (PR) lasting 2+ to 44+ months were seen in 67%; 26% did not respond (NR) and one patient was not evaluable. Hypercalcaemia predicted a poor outcome but differences were not significant. Eight of the 15 patients have died 3–41 months from diagnosis. Median survival for the 15 patients was 18 months. Survival of the NR ranged from 4 to 20 months; six PR patients are alive 8–82 months from diagnosis. The differences in survival between NR (median: 6 months) and PR (55% of patients alive at 4 years) were statistically significant (P = 0·002). In conclusion, IFN and AZT improves the outcome of ATLL patients and helps maintain responses.
Protein Z in ischaemic stroke
Authors: Kobelt Katrin , Biasiutti Franziska Demarmels , Mattle Heinrich P. , Lämmle Bernhard , Wuillemin Walter A.
Published: March 2003

Many risk factors associated with ischaemic stroke are known, including high levels of fibrinogen or factor VII. Protein Z is a vitamin K‐dependent coagulation factor, which was found to promote the assembly of thrombin with phospholipid vesicles that might promote coagulation. Indeed, a low protein Z level may be associated with a varying bleeding tendency. Therefore, we hypothesized that high protein Z levels could induce a hypercoagulable state and performed a case–control study to investigate a potential association between high protein Z plasma levels and ischaemic stroke. We measured protein Z in plasma samples from 157 patients with stroke of unknown aetiology and 192 control subjects. All patients had survived an ischaemic stroke or transient ischaemic attack (TIA) for at least 2 months. We found an increased relative risk of ischaemic stroke with increasing protein Z levels, with an odds ratio of 4·3 [95% confidence interval (CI): 1·7–11] for protein Z plasma levels ≥ 160%. Excluding patients with a history of venous thromboembolism from the analysis, the same result was obtained (odds ratio 4·2; 95% CI: 1·6–11·2). Using a logistic regression model, this association also remained significant (P = 0·04) after adjustment for established risk factors. Our data indicated that a high plasma level of protein Z is an independent risk factor for ischaemic stroke.
T‐cell depletion and autologous stem cell transplantation in the management of tumour stage mycosis fungoides with peripheral blood involvement
Authors: Olavarria Eduardo , Child Fiona , Woolford Alison , Whittaker Sean J. , Davis John G. , McDonald Christine , Chilcott Sarah , Spittle Margaret , J Robert , Grieve  , Stewart Simon , Apperley Jane F. , Russell‐Jones Robin
Published: March 2003

Nine patients with tumour stage mycosis fungoides (MF) have been entered into a pilot study of T‐cell depletion and autologous stem cell transplantation (SCT). Eight patients had detectable rearrangements of the T‐cell receptor (TCR) γ‐gene demonstrated by polymerase chain reaction (PCR)/single‐stranded conformation polymorphism (SSCP) in the peripheral blood. The median age was 47 years and the median duration of disease before SCT was 61 months; Peripheral blood progenitor cells were mobilized using high‐dose etoposide (1·6 g/m2) and granulocyte colony‐stimulating factor (G‐CSF). The apheresis products underwent rigorous T‐cell depletion with immunomagnetic methods. Double CD34‐positive and CD4/CD8‐negative selection achieved a median reduction of 3·89 log of T cells. All nine patients have been transplanted. Conditioning included carmustine (BCNU), etoposide and melphalan (BEM) in seven patients and total body irradiation plus etoposide or melphalan in two. Eight patients engrafted promptly and one patient died of septicaemia. All survivors entered complete remission. Seven patients have relapsed at a median of 7 months (2–14) post SCT. However, most patients have relapsed into a less aggressive stage, which has responded to conventional therapy. Four out of seven evaluable patients had detectable TCR rearrangements in the T‐cell depleted graft. A T‐cell clone was also detected in the peripheral blood before relapse in four cases. Autologous SCT is feasible, safe and can result in complete remission in a significant proportion of patients with tumour stage mycosis fungoides. Despite a short relapse‐free survival, most patients achieved good disease control at the time of relapse.
Improved outcome of alternative donor bone marrow transplantation in children with severe aplastic anaemia using a conditioning regimen containing low‐dose total body irradiation, cyclophosphamide and Campath
Authors: Vassiliou G. S. , Webb D. K. H. , Pamphilon D. , Knapper S. , Veys P. A.
Published: March 2003

The increasing success of human leucocyte antigen (HLA)‐matched sibling donor (MSD) transplants and combination immunosuppressive treatments have dramatically improved the prognosis of severe aplastic anaemia (SAA) in children and young adults. For patients who lack a MSD there is a significant minority who fail immunosuppressive therapy or suffer from a severe constitutional aplastic anaemia in which immunosuppression would be ineffective. Alternative donor bone marrow transplantation (AD‐BMT) has only had limited success in this context. We report the successful outcome of AD‐BMT in eight consecutive patients aged 7 months to 15 years, six of whom had acquired aplastic anaemia who had previously failed to respond to immunosuppression, and two of whom had a severe (non‐Fanconi) constitutional aplastic anaemia. All eight patients had received multiple red cell and platelet transfusions. We used a new combination of agents for pretransplant conditioning aiming to maximize immunosuppression and minimize toxicity, consisting of Campath‐1G or ‐1H, cyclophosphamide and low‐dose total body irradiation (LD TBI) or fludarabine. Toxicity was minimal and all eight children are alive, well and free of disease at a median follow‐up of 32 months. We suggest that this approach could facilitate the successful treatment of children with SAA in whom immunosuppressive therapy has failed or is not appropriate.
Immune function in patients with Shwachman–Diamond syndrome
Authors: Dror Yigal , Ginzberg Hedy , Dalal Ilan , Cherepanov Vera , Downey Gregory , Durie Peter , Roifman Chaim M. , Freedman Melvin H.
Published: March 2003

Shwachman–Diamond syndrome (SDS) is an inherited multisystem disorder characterized by exocrine pancreatic dysfunction and varying degrees of cytopenia. In addition, various immunological abnormalities have been noted. To clarify the issue of immunological competence or incompetence in SDS, we prospectively studied immune function in 11 patients with SDS. Seven suffered from recurrent bacterial infections and six from recurrent viral infections. Varying degrees of impairment were readily identified. All patients had neutropenia; total lymphocyte counts, however, were normal in all except one patient. Nine patients had B‐cell defects comprising one or more of the following abnormalities: low IgG or IgG subclasses, low percentage of circulating B lymphocytes, decreased in vitro B‐lymphocyte proliferation and a lack of specific antibody production. Seven out of nine patients studied had at least one T‐cell abnormality comprising a low percentage of total circulating T lymphocytes or CD3+/CD4+ cell subpopulations or decreased in vitro T‐lymphocyte proliferation. Five out of six patients studied had decreased percentages of circulating natural killer cells. Moreover, neutrophil chemotaxis was significantly low in all the patients studied. These data point to a major immunodeficiency component in SDS that places patients at heightened risk of infections, even if neutrophil numbers are protective. This finding broadens the definition of the syndrome substantially: it suggests that the SDS marrow defect occurs at the level of an early haematological–lymphocytic stem cell or that a combined marrow and thymic stromal defect accounts for the aberrant function of haematopoietic and lymphopoietic lineages.
Increased thromboxane production in women with a history of venous thromboembolic event: effect of heparins
Authors: Kaaja Risto , Pettilä Ville , Leinonen Pekka , Ylikorkala Olavi
Published: March 2003

We investigated the production of prostacyclin and thromboxane in pregnant women with a previous venous thromboembolic event before, during and after the use of unfractionated heparin and low molecular weight heparin (dalteparin). Twenty women were studied before starting heparin prophylaxis (before 20 weeks of gestation), during heparin prophylaxis (at 30 weeks of gestation) and after heparin prophylaxis (16 weeks after delivery). Ten pregnant women with no history of thromboembolism were studied as the control group. Urinary output of the stable metabolite of prostacyclin (2,3‐dinor‐6‐keto‐PGF1α) and that of thromboxane A2 (2,3‐dinor‐TxB2), as well as a number of markers of thrombophilia were measured and expressed as mean (±SEM). Women with a history of thromboembolism were characterized by normal prostacyclin production but elevated thromboxane production (44·0 ± 4·1 versus 19·0 ± 3·6 ng/mmol creatinine, P < 0·001) at 12 weeks of pregnancy. Heparin prophylaxis (regardless of the type) had abolished elevated thromboxane concentrations at 30 weeks of gestation. Four months after delivery, thromboxane dominance had returned (25·2 ± 3·5 versus 13·6 ± 2·1 ng/mmol creatinine, P < 0·01). The presence of hereditary thrombophilia (9/20) was not associated with any changes in prostanoid concentrations. Thus, women with a history of venous thromboembolic events have thromboxane dominance during and after pregnancy, but this dominance can be eliminated through the use of heparins.
Inferior vena cava malformation as a risk factor for deep venous thrombosis in the young
Authors: Chee Yen‐Lin , Culligan Dominic J. , Watson Henry G.
Published: March 2003

Conditions which result in hypercoagulable blood or venous stasis may predispose to the development of deep vein thrombosis (DVT). Most of the recently described risk factors for DVT induce a hypercoagulable state. Over a 3‐year period we have observed anomaly of the inferior vena cava (IVC) in four young patients presenting with spontaneous unprovoked DVT. This is a greater than expected rate (5% observed versus 0·5% expected). Further, bilateral DVT, which constitutes less than 10% of cases in most series, was present in three of the four cases. Anomaly of the IVC is a rare example of a prevalent congenital condition that predisposes to DVT, presumably by favouring venous stasis. This diagnosis should be considered in young patients with spontaneous and bilateral DVT.
Blastic mantle cell lymphoma associated with Burkitt‐type translocation and hypodiploidy
Authors: Vaishampayan Ulka N. , Mohamed Anwar N. , Dugan Michael C. , Bloom Robert E. , Palutke Margarita
Published: March 2003

Two elderly men with stage IV mantle cell lymphoma (MCL) rapidly developed a leukaemic phase with unusually high blast counts that proved fatal. One lymph node biopsy showed diffuse MCL, the other blastic morphology. In addition to t(11;14), there were t(8;14) and t(1;19) in case 1 and dup(8)(q24q13) in case 2. Fluorescence in situ hybridization revealed genomic fusion of IgH/MYC genes in case 1 and an extra copy of C‐MYC gene in case 2. The genomic alteration of C‐MYC oncogene is probably implicated in the blastic transformation and aggressive behaviour of the disease.
Human transferrin G277S mutation: a risk factor for iron deficiency anaemia
Authors: Lee Pauline L. , Halloran Carol , Trevino Renee , Felitti Vincent , Beutler Ernest
Published: March 2003

Numerous polymorphisms of the transferrin gene result in a range of electrophoretic variants. We show that one of these mutations has a functional consequence. A G→A mutation at cDNA nucleotide 829 (G277S) was associated with a reduction in total iron binding capacity (TIBC). In menstruating white women, the G277S genotype was a risk factor for iron deficiency anaemia: iron deficiency anaemia was present in 27% of homozygous G277S/G277S women, 10% of G277G/G277S heterozygous women and 5% of homozygous wild‐type G277G/G277G women.
Dendritic cells in patients with non‐progressive B‐chronic lymphocytic leukaemia have a normal functional capability but abnormal cytokine pattern
Authors: Rezvany Mohammad Reza , Jeddi‐Tehrani Mahmood , Biberfeld Peter , Söderlund Johan , Mellstedt Håkan , Österborg Anders , Rabbani Hodjattallah
Published: March 2003

Dendritic cells (DC) are attractive candidates for use in vaccine‐based immunotherapy. We have analysed the functional capability of DC generated in vitro from blood CD14+ cells of chronic lymphocytic leukaemia (CLL) patients and healthy donors by culturing for 10 d with granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), interleukin 4 (IL‐4) and tumour necrosis factor‐α (TNF‐α). Two distinct DC populations were identified in patients as well as in controls. The majority of DC expressed CD11c and a minority also CD123. Most of the DC generated from both patients and controls exhibited a mature phenotype indicated by CD83 and major histocompatibility complex (MHC) class II expression, as well as by a characteristic morphology. Less than 1% of DC exhibited CD14. CLL DC had a similar expression of accessory molecules (CD54, CD80 and CD86) as control DC. The mean fluorescence intensity of CD80 and MHC class I molecules was significantly higher on CLL DC than on control DC (P < 0·05). At the gene level (real‐time polymerase chain reaction) the expression of IL‐10 was higher in CLL (P = 0·028) than in control DC. IL‐1β and IL‐12p35 transcripts were also more abundant in CLL than in control DC but did not reach statistical significance. The expression of IL‐4 and TNF‐α was similar to that of control DC. The interferon γ (IFN‐γ) gene expression level in CLL DC was decreased compared with control DC. DC of CLL patients had a similar capacity to stimulate in mixed leucocyte reaction as well as to present a recall antigen (PPD) as control DC. Thus, DC of CLL patients seem to have a normal function and may serve as antigen preserving cells for presentation of tumour antigens in a therapeutic vaccination approach. The mechanisms behind the observed increase in some surface molecules and the abnormal cytokine profile of CLL DC is not clear but might indicate pre‐activation of DC in vivo, which may have a regulatory role in the pathobiology of CLL.
Absence of platelet response after eradication of Helicobacter pylori infection in patients with chronic idiopathic thrombocytopenic purpura
Authors: Jarque I. , Andreu R. , Llopis I. , De la Rubia J. , Gomis F. , Senent L. , Jiménez C. , Martín G. , Martínez J. A. , Sanz G. F. , Ponce J. , Sanz M. A.
Published: March 2003

Eradication of Helicobacter pylori infection has been associated with the correction of thrombocytopenia in patients with idiopathic thrombocytopenic purpura (ITP). We have analysed the response to eradication of H. pylori in a series of 56 adult patients with chronic ITP. Forty patients had H. pylori infection (71%) that was eradicated in 23 of 32 evaluable patients (72%). Platelet counts did not significantly vary according to H. pylori treatment outcome. Three of 56 patients (5%) achieved a partial response attributable to H. pylori eradication. Therefore, detection of H. pylori infection should not be routinely included in the initial work‐up of ITP.
Effect of cytokines on growth and differentiation of leukaemic cells with translocation t(6;9)(p23;q34)
Authors: Matsushima Takafumi , Saitoh Takayuki , Karasawa Masamitsu , Takizawa Makiko , Miyawaki Shuichi , Nojima Yoshihisa , Murakami Hirokazu
Published: March 2003

The translocation t(6;9)(p23;q34) is detected infrequently in subtypes of haematological malignancies including acute myelogenous leukaemia (AML) and myelodysplastic syndrome (MDS). Although the t(6;9) leukaemia is commonly associated with bone marrow basophilia, the cytological characteristics of leukaemic cells are unclear. In the current study, we examined the in vitro effects of several cytokines on growth and differentiation of t(6;9) leukaemic cells. Isolated bone marrow mononuclear cells from four patients with t(6;9) (two MDS and two AML) were cultured for 14 d in the presence or absence of each cytokine. At the end of culture, viable cells were counted, and their histology was examined. Bone marrow cells obtained from 22 patients (10 AML, six AML from MDS, six MDS) lacking t(6;9) were used as controls. Compared with control cultures, significantly higher numbers of blasts appeared in the culture of bone marrow cells from t(6;9)‐positive patients in response to stimulation with granulocyte colony‐stimulating factor (G‐CSF), granulocyte–macrophage CSF (GM‐CSF) or interleukin 3 (IL‐3). Stem cell factor (SCF) had little effect. Neutrophil counts were also significantly increased in the presence of G‐CSF or IL‐3. SCF and IL‐3 were potent in increasing basophil counts from t(6;9)‐positive cultures. These findings suggest that bone marrow cells obtained from t(6;9) patients are highly sensitive to growth‐ and/or differentiation‐promoting cytokines. Special attention should be paid to the use of ‘therapeutic’ cytokines in these patients.
Non‐myeloablative bone marrow transplantation in an adult with Wiskott–Aldrich syndrome
Authors: Longhurst Hilary J. , Taussig David , Haque Tanzina , Court Denise Syndercombe , Cavenagh Jamie , Edgar J. David , Helbert Matthew R.
Published: March 2003

Summary. Early bone marrow transplant is now standard treatment for infants with severe immunodeficiencies such as Wiskott–Aldrich Syndrome (WAS), but results in older children and adults are poor. Non‐myeloablative transplant has shown promise in the treatment of older children, who are likely to have active infections and organ damage. We describe a non‐myeloablative transplant of a 26‐year‐old man with WAS, undertaken because of severe infections and vasculitis. Partial engraftment and immunorestoration were achieved. The patient is well 1 year post transplantation.
Results of treatment of children with refractory anaemia with excess blasts (RAEB) and RAEB in transformation (RAEBt) in Great Britain 1990–99
Authors: Webb David K. H. , Passmore Sarah J. , Hann Ian M. , Harrison Georgina , Wheatley Keith , Chessells Judith M.
Published: March 2003

Summary. Between 1990 and 1999, 36 children with refractory anaemia with excess blasts (RAEB) and RAEB in transformation (RAEBt), not associated with Down's syndrome, were diagnosed in Britain. A total of 31 children received intensive chemotherapy, six of whom proceeded to a bone marrow allograft in first remission, whereas two received an autograft. Of the 23 given chemotherapy only, four died of toxicity, 10 relapsed and nine are alive in first remission. Out of the 10 who relapsed, four are alive and disease‐free following an allograft. Out of the 6 children given an allograft in first remission, two died of disease and four are alive in first remission. Both children given an autograft died of disease. Two children received an allograft without prior chemotherapy but died of toxicity. Three children received supportive care only, and one child survived. The overall survival was 51% at 5 years, and was superior in children with RAEBt (63%) compared with RAEB (28%, P = 0·03). Cytogenetics were available in 35 cases. Monosomy 7 was the most common abnormality (33% of cases). Survival in children with monosomy 7 was 22% at 5 years compared with 66% for the other patients (P = 0·05). Allowing for cytogenetics, outcomes of therapy appear similar to those for de novo acute myeloid leukaemia (AML), and it is appropriate for children with RAEB/RAEBt to be registered in AML trials.
Abnormal telomere shortening in leucocytes of children with Shwachman–Diamond syndrome
Authors: Thornley Ian , Dror Yigal , Sung Lilian , Wynn Rob F. , Freedman Melvin H.
Published: March 2003

Summary.  Haemopoietic dysfunction, ranging from single‐lineage cytopenia to severe aplasia and/or myelodysplasia (MDS), is prominent in Shwachman–Diamond syndrome (SDS). To assess haemopoietic stem cell proliferation in SDS, we compared leucocyte telomere length in 12 patients with SDS to that of 41 controls, using an in‐gel hybridization technique. SDS patients had an age‐adjusted mean telomere length 1·4 kilobase pairs (kbp) shorter than controls (P < 0·0001). Patients with'non‐severe′ SDS (one‐ or two‐lineage cytopenias; no MDS) had shortened telomeres (−1·4 kbp; P = 0·0004), as did those with ‘severe’ SDS. We conclude that stem cell hyperproliferation is a feature of SDS from its outset.
Inherited factor VII deficiency and surgery: clinical data are the best criteria to predict the risk of bleeding
Authors: Blaizot Muriel Giansily , Biron‐Andreani Christine , Aguilar‐Martinez Patricia , De Moeloose Philippe , Briquel Marie‐Elisabeth , Goudemand Jenny , Stieltjes Nathalie , Barrot Claire , Chambost Herve , Durin Anne , Gay Valerie , Peynet Jocelyne , Pouymayou Katia , Schved Jean‐François
Published: March 2003

Summary.Inherited factor VII (FVII) deficiency is a rare autosomal disorder characterized by a weak relationship between FVII activity (FVII:C) and operative bleeding risk. We report a retrospective study of 17 patients with a FVII:C below 0·1 IU/ml, in whom surgery was performed without any replacement therapy. Clinical and biological data were analysed to establish predictive criteria for bleeding tendency. We found that systematic preoperative replacement therapy may not be necessary for ‘minor’ surgical procedures, for patients suffering from inherited FVII deficiency, unless the clinical history includes severe haemorrhagic symptoms such as haemarthrosis, severe haematomas (even of soft tissue) or abundant epistaxis.
Effect of VDR polymorphisms on growth and bone mineral density in homozygous beta thalassaemia
Authors: Ferrara Mara , Matarese Sofia M. R. , Francese Matteo , Borrelli Barbara , Coppola Antonietta , Coppola Lina , Esposito Luigi
Published: March 2003

Summary. We examined the effect of vitamin D receptor (VDR) polymorphisms at exon 2 (FokI) and intron 8 (BsmI) on the stature and bone mineral density at femoral neck (FBMD) and lumbar spine (LBMD) in 108 prepubertal and pubertal homozygous β thalassaemic patients, regularly treated. We found significantly shorter stature and lower LBMD and FBMD in all patients with CC VDR genotype, and significant shorter height and lower LBMD in prepubertal and pubertal female patients with BB VDR genotype. Because homozygous CC and BB VDR genotypes influence Vitamin D activity, they can be considered additional risk factors for bone disease in β thalassaemia.
Reduced platelet adhesion in flowing blood to fibrinogen by alterations in segment γ316–322, part of the fibrin‐specific region*
Authors: Remijn Jasper A. , IJsseldijk Martin J. W. , Van Hemel Bettien M. , Galanakis Dennis K. , Hogan Kelly A. , Lounes Karim C. , Lord Susan T. , Sixma Jan J. , De Groot Philip G.
Published: March 2003

Summary. The interaction of platelets with fibrinogen is a key event in the maintenance of a haemostatic response. It has been shown that the 12‐carboxy‐terminal residues of the γ‐chain of fibrinogen mediate platelet adhesion to immobilized fibrinogen. These studies, however, did not exclude the possibility that other domains of fibrinogen are involved in interactions with platelets. To obtain more insight into the involvement of other domains of fibrinogen in platelet adhesion, we studied platelet adhesion in flowing blood to patient dysfibrinogen Vlissingen/Frankfurt IV (V/FIV), to several variant recombinant fibrinogens with abnormalities in the γ‐chain segments γ318–320 and γ408–411. Perfusion studies at physiological shear rates showed that platelet adhesion was absent to γΔ408‐411, slightly reduced to the heterozygous patient dysfibrinogen V/FIV and strongly reduced to the homozygous recombinant fibrinogens: γΔ319‐320, γ318Asp→Ala and γ320Asp→Ala. Furthermore, antibodies raised against the sequences γ308–322 and γ316–333 inhibited platelet adhesion under shear conditions. These experiments indicated that the overlapping segment γ316–322 contains amino acids that could be involved in platelet adhesion to immobilized fibrinogen under flow conditions. In soluble fibrinogen, this sequence is buried inside the fibrinogen molecule and becomes exposed after polymerization. In addition, we have shown that this fibrin‐specific sequence also becomes exposed when fibrinogen is immobilized on a surface.
DDAVP challenge tests in boys with mild/moderate haemophilia A*
Authors: Revel‐Vilk Shoshana , Blanchette Victor S. , Sparling Cathy , Stain Ann Marie , Carcao Manuel D.
Published: March 2003

Summary. Desmopressin (DDAVP) increases plasma factor VIII coagulant activity (FVIII:C) levels in patients with mild/moderate haemophilia A. In some subjects, FVIII can be increased to haemostatic levels, thereby avoiding use of factor VIII concentrates. We reviewed our hospital's experience with 62 boys with FVIII:C levels 0·01–0·3 IU/ml who had a DDAVP challenge test (i.v. 0·3 µg/kg) following diagnosis. A therapeutic response was defined as a 1 h post‐FVIII:C increase at least twofold over baseline and > 0·3 IU/ml. Of the total group, 29 (47%) boys responded to DDAVP, all of them with mild haemophilia (baseline FVIII:C ≥ 0·05 IU/ml), yielding a response rate of 57% in this subgroup. Boys who responded to DDAVP had higher baseline FVIII:C levels (mean ± SEM, 0·17 ± 0·01 vs 0·10 ± 0·01 IU/ml, P < 0·01) and were older (5·2 ± 0·8 vs 3 ± 0·4 years, P = 0·02) than those who failed to do so. The association between DDAVP response and age, however, remains unclear: seven boys who failed the initial challenge test responded to re‐challenge after a mean of 6·3 years (median 4·9, range 0·5–12·5), increasing the response rate in boys with mild haemophilia to 71%. Age and FVIII:C association with DDAVP response are both important in boys with mild/moderate haemophilia A. Absence of response to DDAVP should therefore be confirmed by later re‐challenge.