bluuebunny/crossref_metadata_2025 huggingface dataset:
Allele and haplotype frequency at human leucocyte antigen class I/II and immunomodulatory cytokine loci in patients with myelodysplasia and acute myeloid leukaemia: in search of an autoimmune aetiology
Authors: Gowans Duncan , O'Sullivan Alan , Rollinson Sara , Roddam Phillipa , Groves Mike , Fegan Chris , Morgan Gareth , Bowen David
Published: March 2003

Summary.  An autoimmune mechanism in the␣pathogenesis of myelodysplastic syndrome (MDS) is suggested by response to immunosuppression, with CD8+ T‐lymphocytes implicated in the haematopoietic suppression. We therefore sought evidence for human leucocyte antigen (HLA) restriction and variant frequency differences in selected polymorphisms at␣the loci for the immunomodulatory cytokines, tumour necrosis factor α (TNF‐α), lymphotoxin‐α (LT‐α) and interleukin 10 (IL‐10) in patients with MDS and acute myeloid leukaemia (AML) compared with normal controls. DNA from 150 MDS/AML patients [24 AML, 53 refractory anaemia (RA), 25 RA with excess blasts (RAEB), four RAEB in transformation (RAEBt), 21 sideroblastic leukaemia, 22 chronic myelomonocytic leukaemia] was screened. Control data was from Scottish blood donors (HLA class I/II), healthy General Practitioner‐based subjects (TNF‐α/LT‐α) and published values (IL‐10). HLA class I/II haplotypes were determined using sequence‐specific primers. Polymorphisms were assayed at TNF‐α−308, LT‐α +252 and IL10 −824, −597 and −1082 loci. Variant frequencies of common haplotypes at HLA class I and II, high‐/low‐producer TNF‐α/LT‐α and IL‐10 loci were not different between patients and controls or within the French–American–British, International Prognostic Scoring System or cytogenetic subgroups and were not associated with altered survival for MDS/AML patients. TNF2 allele frequency was greater in the MDS/AML cohort (χ2 = 6·593, P < 0·05) but the biological significance was uncertain in the absence of an increased high‐producer TNF‐α/LT‐α haplotype frequency. We can find no genetic influence for these polymorphisms in HLA class I/II, TNF‐α/LT‐α and IL‐10 loci on either predisposition or disease progression in MDS/AML.
Early lymphocyte recovery post‐autologous haematopoietic stem cell transplantation is associated with better survival in Hodgkin's disease
Authors: Porrata Luis F. , Inwards David J. , Micallef Ivana N. , Ansell Stephen M. , Geyer Susan M. , Markovic Svetomir N.
Published: March 2003

Summary.A retrospective study of 82 patients was conducted to determine the relationship of absolute lymphocyte count (ALC) recovery with clinical outcome after autologous stem cell transplantation (ASCT) in Hodgkin's disease (HD). The median overall (OS) and progression‐free survival (PFS) times from the day of transplantation were significantly better for the 41 patients with ALC ≥ 0·5 × 109cells/l compared with the 41 patients with ALC < 0·5 × 109cells/l (‘not yet reached’ versus 42 months,P < 0·0001; 57 versus 15 months,P < 0·002 respectively). Thus, ALC recovery on day 15 post ASCT in HD is associated with better survival and requires further study.
Trisomy 3 in two paediatric post‐transplant lymphomas
Authors: Gallego Marta S. , Bernasconi Andrea , Dávila María T. , Rampazzi Alejandra , Chantada Guillermo , Gallo Guillermo , Rossi Jorge , Barreiro Cristina Z.
Published: March 2003

Summary. Few cytogenetic data are available concerning the chromosomal constitution of post‐transplant lymphomas. We report two paediatric cases of trisomy 3, as a primary anomaly, in post‐transplant lymphoproliferative disease (PTLD) associated with B immunophenotype. Using cytogenetic analysis and fluorescence in situ hybridization on chromosome preparations, we found trisomy 3 in both patients and an extra X chromosome in one. Clinical, histological and immunophenotypical data are presented. Trisomy 3 has been observed in different types of non‐Hodgkin's lymphomas but it is relatively rare in B‐cell lymphomas, with the exception of marginal zone lymphoma and mantle cell lymphoma. To our knowledge, trisomy 3 is an uncommon cytogenetic finding in PTLD. Further cytogenetic studies of these lymphoproliferative disorders might contribute to evaluate the role of these chromosomal anomalies in the pathogenesis of this disease.
Stevens–Johnson syndrome after treatment with STI571: a case report
Authors: Hsiao Liang‐Tsai , Chung Hung‐Ming , Lin Jen‐Tsun , Chiou Tzeon‐Jye , Liu Jin‐Hwang , Fan Frank S. , Wang Wei‐Shu , Yen Chueh‐Chuan , Chen Po‐Min
Published: March 2003

Summary.  Between seven and 21% of patients treated with the specific tyrosine kinase inhibitor STI571 have been reported to develop mild‐to‐moderate severity of adverse cutaneous reactions. We report a patient in the blast crisis phase of chronic myeloid leukaemia who developed a life‐threatening cutaneous reaction, Stevens–Johnson syndrome, following 1 week of STI571 therapy. This report may serve to remind the clinician about the possible severe cutaneous side‐effects of STI571 before instituting more extensive clinical application of this agent in the future.
Platelet‐derived growth factor promotes ex vivo expansion of CD34+ cells from human cord blood and enhances long‐term culture‐initiating cells, non‐obese diabetic/severe combined immunodeficient repopulating cells and formation of adherent cells
Authors: Su Rui Jun , Zhang Xiao Bing , Li Karen , Yang Mo , Li Chi Kong , Fok Tai Fai , James Anthony Edward , Pong Henry , Yuen Patrick Man Pan
Published: March 2003

Summary. Platelet‐derived growth factor (PDGF) is a major mitogen for connective tissue cells. In this study, we investigated the effects and mechanism of PDGF on the ex vivo expansion of cord blood CD34+ cells. Our data demonstrated that among various cytokine combinations of thrombopoietin (TPO), interleukin 1 beta (IL‐1β), IL‐3, IL‐6 and Flt‐3 ligand (Flt‐3L), TPO + IL‐6 + Flt‐3L was most efficient in promoting the expansion of CD34+ cells, CD34+CD38– cells, mixed‐lineage colony‐forming units (CFU‐GEMM) and long‐term culture‐initiating cells (LTC‐IC) by 21·7 ± 5·00‐, 103 ± 27·9‐, 10·7 ± 7·94‐ and 6·52 ± 1·51‐fold, respectively, after 12–14 d of culture. The addition of PDGF increased the yield of these early progenitors by 45·0%, 66·5%, 45·1% and 79·8% respectively. More significantly, PDGF enhanced the engraftment of human CD45+ cells and their myeloid subsets (CD33+, CD14+ cells) in non‐obese diabetic (NOD)/severe‐combined immunodeficient (SCID) mice. The expression of PDGF receptor (PDGFR)‐β was not detectable in fresh CD34+ cells but was upregulated after culture for 3 d. PDGF also enhanced the development of adherent cells/clusters that expressed the endothelial markers VE‐cadherin and CD31. These findings suggest that PDGF is an effective cytokine for the ex vivo expansion of early stem and progenitor cells. The mechanism could be mediated by PDGFR‐β on committed CD34+ progenitor cells and/or secondary to the stimulation of autologous, stromal feeder cells.
Time to platelet recovery predicts outcome of patients withde novoacute lymphoblastic leukaemia who have achieved a complete remission
Authors: Faderl Stefan , Thall Peter F. , Kantarjian Hagop M. , Estrov Zeev
Published: March 2003

Summary.Survival in acute leukaemia depends on the achievement of complete remission (CR). However, CR is not a clear‐cut phenomenon and certain variables of its definition could more accurately characterize the quality of the remission. Because platelet recovery > 100 × 109/l is an essential component of CR in acute leukaemia, we hypothesized that time to platelet recovery (TPR) might be predictive of overall survival (OS) or disease‐free survival (DFS) in acute lymphoblastic leukaemia (ALL). We analysed TPR in 249 patients with ALL who entered CR after one course of induction chemotherapy and correlated TPR with DFS and OS. TPR was significantly associated with both DFS and OS if it occurred within a maximum of about 60 d from start of therapy. Furthermore, during that time period, the relative risk of death increased with increasing TPR. Although presence of the Philadelphia chromosome was the single most important adverse feature at diagnosis, the effect of TPR on survival continued to be significant within this patient subgroup. This effect was so pronounced that Philadelphia chromosome‐positive patients with a TPR of 12 d had a better outcome than Philadelphia chromosome‐negative patients with a TPR of 48 d. Thus, a short TPR seems to be able to override adverse characteristics in the outcome of ALL patients treated with chemotherapy. We conclude that a quicker TPR predicts longer DFS and OS in patients with ALL. As platelet counts are obtained almost daily in patients undergoing chemotherapy, TPR can readily be utilized to assess the prognosis of these patients.
Therapy‐related chronic myelogenous leukaemia following autologous stem cell transplantation for Ewing's sarcoma
Authors: Numata Akihiko , Shimoda Kazuya , Gondo Hisashi , Kato Kouji , Aoki Kenichi , Ito Yoshikiyo , Takase Ken , Asano Yoshinobu , Okamura Takashi , Niho Yoshiyuki , Harada Mine
Published: March 2003

Summary. A 17‐year‐old Japanese woman with Ewing's sarcoma was initially treated with conventional chemotherapy and local irradiation, and then with high‐dose chemotherapy supported by autologous peripheral blood stem cell transplantation. Four years later she was diagnosed with chronic myelogenous leukaemia (CML). The BCR/ABL fusion gene was detected in both peripheral blood and bone marrow cells by reverse transcription–polymerase chain reaction, but not in the harvest product of peripheral blood stem cells which were infused at the time of transplantation. This case adds to the accumulating evidence of therapy‐related CML developing after high‐dose chemotherapy and autologous stem cell transplantation.
Dose escalation therapy in previously untreated patients with multiple myeloma following Z‐Dex induction treatment
Authors: Clark Andrew D. , Douglas Kenneth W. , Mitchell Lyndsay D. , McQuaker I. Grant , Parker Anne N. , Tansey Patrick J. , Franklin Ian M. , Cook Gordon
Published: March 2003

Summary. A phase I–II study of high‐dose (HD) alkylating agents in newly diagnosed patients with multiple myeloma after maximum response to Z‐Dex (idarubicin, dexamethasone) therapy and DHAP (cisplatin, HD cytosine arabinoside, dexamethasone), stem cell mobilization is reported. Twenty‐six patients, median age 56 years (range 42–66), completed Z‐Dex chemotherapy and peripheral blood stem cells (PBSC) were mobilized with DHAP. Patients then preceded to cyclophosphamide (HD Cy: 6 g/m2) with granulocyte colony‐stimulating factor followed by busulphan–melphalan‐conditioned PBSC autograft. Interferon α was introduced at 3 months post transplant as maintenance therapy. Six patients failed to complete the full protocol. Median time from diagnosis to transplantation was 8 months (range 6–12). Mean CD34+ cell dose collected was 15·8 × 106/kg (CI 11·8, 19·8). Median time from DHAP to HD‐Cy was 6 weeks (range 4–12) and from HD‐Cy to transplant was 8 weeks (range 6–12). The median follow‐up was 36 months (range 6–63). On an intent‐ to‐treat basis, the response rates were three complete response (CR, 12%), 21 partial response (PR, 80%) and two stable disease (SD, 8%) post Z‐Dex, five CR (19%) and 21 PR (81%) post HD‐Cy, and 14 CR (54%) and 12 PR (46%) post transplant. The treatment‐related mortality (TRM) was 4% (1 patient). Median overall survival (OS) and progression‐free survival (PFS) have not been reached; estimated values were 60 and 48 months respectively. The 3‐year OS and PFS were 72% and 62%. Actuarial 5‐year OS and event‐free survival were 49% and 32%. DHAP produces effective PBSC mobilization and sequential HD therapy, including autologous PBSCT, in patients who received Z‐Dex; this offers significant durable disease response rates with acceptable TRM.
Analysis of resveratrol‐induced apoptosis in human B‐cell chronic leukaemia
Authors: Roman Viviana , Billard Christian , Kern Catherine , Ferry‐Dumazet Hélène , Izard Jean‐Claude , Mohammad Ramzi , Mossalayi Djavad M. , Kolb Jean‐Pierre
Published: March 2003

Summary.  Trans‐resveratrol was analysed for its apoptotic and growth inhibitory activity in human B‐cell lines derived from chronic B‐cell malignancies (WSU‐CLL and ESKOL), and in leukaemic lymphocytes from patients with B‐cell chronic lymphocytic leukaemia (B‐CLL). Resveratrol displayed antiproliferative activity on both B‐cell lines, as estimated by the decrease in cell recovery and inhibition of thymidine uptake. Furthermore, resveratrol induced apoptosis in the two cell lines as well as in B‐CLL patients' cells, as evidenced by the increase in annexin V binding, caspase activation, DNA fragmentation and decrease of the mitochondrial transmembrane potential ΔΨm. We previously reported that nitric oxide (NO), endogenously released by an iNO synthase (iNOS) spontaneously expressed in these leukaemic cells, contributed to their resistance towards apoptosis. We show here that resveratrol inhibited both iNOS protein expression and in situ NO release in WSU‐CLL, ESKOL and B‐CLL patients'cells. In addition, Bcl‐2 expression was also inhibited by resveratrol. Thus, downregulation of the two anti‐apoptotic proteins iNOS and Bcl‐2 can contribute to the apoptotic effects of resveratrol in leukaemic B cells from chronic leukaemia. Our data suggest that this drug is of potential interest for the therapy of B‐CLL.
In vivo mechanisms of resistance to cytarabine in acute myeloid leukaemia
Authors: Galmarini Carlos M. , Thomas Xavier , Calvo Fabien , Rousselot Philippe , Rabilloud Muriel , El Jaffari Assia , Cros Emeline , Dumontet Charles
Published: March 2003

Summary.  Factors that reduce the intracellular concentration of triphosphorylated cytarabine (ara‐CTP), the active form of cytarabine (ara‐C), may induce chemoresistance in acute myeloid leukaemia (AML) patients. These factors include reduced influx of ara‐C by the hENT1 transporter, reduced phosphorylation by deoxycytidine kinase (dCK), and increased degradation by high Km cytoplasmic 5′‐nucleotidase (5NT) and/or cytidine deaminase (CDD). Increased levels of DNA polymerase α (DNA POL) and reduced levels of topoisomerase I (TOPO I) and topoisomerase II (TOPO II) have also been detected in ara‐C‐resistant cell lines. To determine whether these factors are implicated in clinical ara‐C resistance, we analysed the expression of these parameters at diagnosis, using reverse transcription polymerase chain reaction, in the blast cells of 123 AML patients treated with ara‐C. At diagnosis, hENT1, dCK, CDD, 5NT, TOPO I, TOPO II, DNA POL and MDR1 were expressed in 83%, 22%, 7%, 37%, 59%, 37%, 39% and 16% of patients respectively. In univariate analysis, patients with expression of 5NT or DNA POL at diagnosis had significantly shorter disease‐free survival (DFS). In multivariate analysis, DNA POL positivity and hENT1 deficiency were related to a shorter DFS. In univariate analysis, patients with 5NT‐positive blasts had significantly shorter overall survival (OS). In multivariate analysis, shorter OS was related to DNA POL positivity. These results suggest that expression of DNA POL, 5NT and hENT1 at diagnosis may be resistance mechanisms to ara‐C in AML patients.
Distinct genetic patterns can be identified in acute monoblastic and acute monocytic leukaemia (FAB AML M5a and M5b): a study of 124 patients
Authors: Haferlach Torsten , Schoch Claudia , Schnittger Susanne , Kern Wolfgang , Löffler Helmut , Hiddemann Wolfgang
Published: March 2003

Summary. The French–American–British (FAB) classification and the new World Health Organization (WHO) classification distinguish acute monoblastic leukaemia (AML M5a) from acute monocytic leukaemia (AML M5b). Not much is known about the underlying genetic differences leading to these clearly different phenotypes. We analysed 58 patients with de novo AML M5a and 66 patients with denovo AML M5b in comparison with a whole group of 1603 de novo AML. An aberrant karyotype was found in 75·9% of AML M5a but in only 28·8% of M5b (P < 0·0001) and in 54·7% of all other AML subtypes (P = 0·0015). 11q23/MLL aberrations were detected in 31% of M5a, 12·1% of M5b (P = 0·01) but only 1·3% of all other AML subtypes (P < 0·0001). Trisomy 8 as the sole cytogenetic aberration was found in 22·4% of M5a, but in only 3% of M5b and in 2·5% of all other AML subcategories (P < 0·0001). Although the frequency of the MLL‐partial tandem duplication (MLL‐PTD) did not differ between the three cohorts (1·7%, 4·5% and 6·1% respectively, NS), thedetection of FLT3 length mutations (FLT3‐LM) differed significantly. AML M5a showed a low frequency of only 6·9%, but 28·8% of M5b (P = 0·0014) and 23·5% of all other AML revealed a FLT3‐LM. In conclusion, we demonstrated genetic, i.e. biological, differences between AML M5a and AML M5b and all other AML. Therefore, AML M5 should further be categorized as two different groups, as proposed by the WHO classification.
The role of strain gauge plethysmography in the assessment of patients with suspected deep vein thrombosis
Authors: Robinson Brian J. , Kesteven Patrick J. L. , Elliott Simon T.
Published: March 2003

Summary. Suspicion of deep vein thrombosis (DVT) is a common reason for medical referral to hospital. Clinical signs and symptoms are notoriously unreliable, hence there is the need for objective testing. Strain gauge plethysmography (SGP) has been marketed as a technique for excluding lower limb DVT. We therefore set out to evaluate this screening tool. Over a 2 year period, 437 consecutive patients referred with suspected DVT were assessed using both plethysmography and Doppler ultrasound. When the two techniques were compared, plethysmography was found to have a negative predictive value of 90%. We conclude that strain gauge plethysmography has a role in the screening of patients with suspected DVT but should not be used as the sole method in patient assessment.
β2‐microglobulin as a negative growth regulator of myeloma cells
Authors: Min Rui , Li Zhongkui , Epstein Joshua , Barlogie Bart , Yi Qing
Published: March 2003

Summary. High β2‐microglobulin (β2m) levels in myeloma correlate with poor prognosis. We hypothesized that β2m may affect myeloma cell growth and survival. In this study, we examined the in vitro effects of β2m on myeloma cells. Primary myeloma cells freshly isolated from patients and myeloma cell lines were used, cultured in the presence of β2m, and monitored for growth and survival. β2m suppressed the growth of primary tumour cells and myeloma cell lines (ARK‐RS, ARP‐1, RPMI‐8226, U266, ARH‐77 and IM‐9). High concentrations of β2m induced apoptosis and cell cycle arrest. β2m‐induced apoptosis was dependent on activation of a caspase cascade, inhibited by interleukin 6, and did not involve the surface death receptors, as receptor‐neutralizing antibodies had no inhibitory effect. β2m‐induced growth arrest was associated with downregulation of cyclins A and D2. Surprisingly, anti‐β2m antibodies did not block the effect of β2m but were synergistic with β2m, resulting in 90% growth inhibition and 70% apoptosis of myeloma cells. Whereas β2m treatment resulted in slight upregulation of surface β2m and major histocompatibility complex class I α‐chain expression, treatment of myeloma cells with anti‐β2m antibodies alone or with β2m resulted in significant downregulation of surface β2m and class I molecules, suggesting that class I molecules may be involved in signal transduction. Our data demonstrate that β2m plays an important role in regulating the growth and survival of myeloma cells in vitro and warrants further investigation to delineate the mechanisms of β2m and anti‐β2m antibody‐induced growth regulation of myeloma cells.
Long‐term follow up with conventional cytogenetics and band 13q14 interphase/metaphase in situ hybridization monitoring in monoclonal gammopathies of undetermined significance
Authors: Bernasconi Paolo , Cavigliano Paola Maria , Boni Marina , Astori Cesare , Calatroni Silvia , Giardini Ilaria , Rocca Barbara , Caresana Marilena , Crosetto Nicola , Lazzarino Mario , Bernasconi Carlo
Published: March 2003

Summary. One‐third of patients with monoclonal gammopathy of undetermined significance (MGUS) may progress to multiple myeloma (MM) and may develop a long arm deletion of chromosome 13 (13q–). As the incidence of 13q–, time of development and prognostic impact in MGUS patients is still under debate, we decided to perform serial sequential conventional cytogenetics (CC) and metaphase/interphase fluorescence in situ hybridization (FISH) analyses on bone marrow mononuclear cells obtained from 18 asymptomatic, untreated MGUS patients. Median follow up was 30 months (range 6–72). Interphase FISH identified a 13q14 deletion in five out of 18 patients (on clinical diagnosis in one patient and during the follow up in the remaining four patients). Subsequently, metaphase FISH and CC also identified the deletion in four out of five patients. All five of the patients progressed to MM 6–12 months after 13q– identification, without developing any FISH determined JH rearrangements. MM progression also occurred in two other karyotypically normal patients. We conclude that: (i) the extent of the 13q deletion does not vary during the clinical outcome; (ii)13q– plays a crucial role in MGUS/MM pathogenesis and confers a proliferative advantage to clonal plasma cells being initially demonstrated by interphase FISH and only afterwards by metaphase FISH and CC; and (iii) association of 13q– with t(4;14)(p16.3;q32) remains to be demonstrated. However, a transition from MGUS to MM may also occur in patients with normal karyotypes or other abnormalities, suggesting the possibility of distinct pathogenetic pathways.
Enforced expression of HOX11 is associated with an immature phenotype in J2E erythroid cells
Authors: Greene Wayne K. , Ford Jette , Dixon Darcelle , Tilbrook Peta A. , Watt Paul M. , Klinken S. Peter , Kees Ursula R.
Published: March 2003

Summary. The HOX11 gene encodes a homeodomain transcription factor that is essential for spleen development during embryogenesis. HOX11 is also leukaemogenic, both through its clinical association with childhood T‐cell acute lymphoblastic leukaemia, and its ability to immortalize other haematopoietic cell lineages experimentally. To examine the pathological role of HOX11 in tumorigenesis, we constitutively expressed HOX11 cDNA in J2E murine erythroleukaemic cells, which are capable of terminal differentiation. Enforced HOX11 expression was found to induce a profound alteration in J2E cellular morphology and differentiation status. Our analyses revealed that HOX11 produced clones with a preponderance of less differentiated cells that were highly adherent to plastic. Morphologically, the cells overexpressing HOX11 were larger and had decreased globin levels, as well as a reduction in haemoglobin synthesis in response to erythropoietin (EPO). Immunocytochemical analysis confirmed the immature erythroid phenotype imposed by HOX11, with clones transfected with HOX11 demonstrating expression of the c‐Kit stem cell marker, while retaining EPO receptor expression. Taken together, these results show that HOX11 alters erythroid differentiation, favouring a less mature progenitor‐like stage. This supports the notion that disrupted haematopoietic cell differentiation is responsible for pre‐leukaemic immortalization by the HOX11 oncoprotein.
Natural cytotoxicity to autologous antigen‐pulsed dendritic cells in multiple myeloma
Authors: Zheng Chengyun , Ostad Masih , Andersson Margareta , Celsing Fredrik , Holm Göran , Sundblad Anne
Published: March 2003

Summary. To analyse autologous lymphocyte cytolytic activities of potential importance for cell‐based immunotherapy in multiple myeloma (MM), in vitro differentiated dendritic cells (DCs) loaded with patient‐specific monoclonal immunoglobulin (mIg) were used as autologous target cellsin cytotoxicity assays. Effector populations consisted of purified natural killer (NK) cells (CD56+, CD3–) and T cells (CD3+). The MM patients' NK cells cultured in the presence of interleukin 2 (IL‐2) showed pronounced cytotoxic activity towards autologous mature DCs. Autologous MM DC targets displayed similar susceptibility to NK cell lysis, compared with allogeneic control DC targets, despite high surface expression of self major histocompatibility complex (MHC) antigens. However, some degree of classic MHC class I‐mediated negative regulation was implicated in the NK–DC interactions, as indicated by class I blocking experiments. NK‐mediated lysis was also discerned towards primary autologous MM cells. The results indicated that the major effector mechanism was mediated through the perforin–granzyme exocytosis pathway. In conclusion, NK cells from MM patients displayed significant and consistent cytotoxicity towards autologous mature DCs, suggesting that innate immunity could be implicated in MM and may influence the outcome of the administration of tumour antigen‐pulsed DCs in treatment trials.
Risk‐directed treatment of infant acute lymphoblastic leukaemia based on early assessment ofMLLgene status: results of the Japan Infant Leukaemia Study (MLL96)
Authors: Isoyama Keiichi , Eguchi Mariko , Hibi Shigeyoshi , Kinukawa Naoko , Ohkawa Hiroji , Kawasaki Hajime , Kosaka Yoshiyuki , Oda Takanori , Oda Megumi , Okamura Takayuki , Nishimura Shin‐ichiro , Hayashi Yasuhide , Mori Taijiro , Imaizumi Masue , Mizutani Shuki , Tsukimoto Ichiro , Kamada Nanao , Ishii Eiichi
Published: March 2003

Summary. We studied the effectiveness of risk‐directed therapy for infants younger than 13 months of age with acute lymphoblastic leukaemia (ALL). Fifty‐five infants were assigned to different treatment programs (from December 1995 to December 1998) on the basis of theirMLLgene status at diagnosis. Forty‐two cases (76·3%) had a rearrangedMLLgene (MLL+) and were treated with remission induction therapy followed by sequential intensive chemotherapy, including multiple genotoxic agents (MLL9601 protocol). Haematopoietic stem cell transplantation (HSCT) was attempted if suitable donors were available. Thirteen infants (23·7%) were classified asMLL–and treated for 2·5 years with intensive chemotherapy for high‐risk B‐ALL (MLL9602 protocol). Complete remission was induced in 38 of the 42 infants (90·5%) withMLL+ALL and in all 13 patients (100%) withMLL–disease. In theMLL+subgroup, the estimated event‐free survival (EFS) rate at 3 years post diagnosis was 34·0% ± 7·5%, compared with 92·3% ± 7·4% in theMLL–subgroup (overall comparison,P = 0·001 by log‐rank analysis). Both age less than 6 months (hazard ratio = 6·87, 95% CI = 0·91–52·3;P = 0·013) and central nervous system (CNS) involvement at diagnosis (hazard ratio = 2·92 95% CI = 1·29–6·63;P = 0·015) were significant independent predictors of an inferior outcome. These findings indicate a strategic advantage in classifying infant ALL as eitherMLL+orMLL–early in the clinical course and selecting therapy accordingly. Standard chemotherapy for high‐risk B‐lineage ALL appeared adequate forMLL–cases. Novel therapeutic initiatives are warranted for infants withMLL+disease, particularly those with initial CNS leukaemic involvement or age less than 6 months, or both.
Mast cell infiltration correlates with poor prognosis in Hodgkin's lymphoma
Authors: Molin Daniel , Edström Annika , Glimelius Ingrid , Glimelius Bengt , Nilsson Gunnar , Sundström Christer , Enblad Gunilla
Published: March 2003

Summary. Hodgkin's lymphoma (HL) is characterized by a few Hodgkin, Reed–Sternberg cells (HRS) surrounded by benign cells. We recently reported that mast cells were the predominant CD30L‐positive cells in HL tumours, and that they activate HRS in vitro through CD30L–CD30 interaction. Here, we investigated the clinical importance of mast cell infiltration in the tumours of 123 patients. Tumour specimens were stained with a mast‐cell‐specific antibody that detects tryptase. Mast cells were detected in virtually every case and increasing numbers of mast cells correlated to nodular sclerosis histology (P = 0·008). Patients with higher mast cell infiltration had a worse relapse‐free survival (P = 0·01).
Severe methylenetetrahydrofolate reductase deficiency revealed by a pulmonary embolism in a young adult
Authors: Tonetti Carole , Ruivard Marc , Rieu Virginie , Zittoun Jacqueline , Giraudier Stephane
Published: March 2003

Summary. Deficiency in methylenetetrahydrofolate reductase (MTHFR), the enzyme involved in the remethylation of homocysteine to methionine using methyltetrahydrofolate as cofactor, induces hyperhomocysteinaemia, homocysteinuria, hypomethioninaemia and low methylfolate levels. Diagnosis usually occurs during infancy because of various neurological abnormalities. We report MTHFR deficiency diagnosed in an adult woman after a pulmonary embolism. Her adult sister, intellectually retarded, suffered from the same disease. Molecular analysis of the MTHFR gene exhibited four different mutations (two missense mutations, one exon skipping and C677T). The impact of these mutations was analysed through the biological abnormalities in the parents and children.
Double‐chimaerism after transplantation of two human leucocyte antigen mismatched, unrelated cord blood units
Authors: De Lima Marcos , St. John Lisa S. , Wieder Eric D. , Lee Ming S. , McMannis John , Karandish Safa , Giralt Sergio , Beran Miroslav , Couriel Daniel , Korbling Martin , Bibawi Samer , Champlin Richard , Komanduri Krishna V.
Published: March 2003

Summary. The small number of progenitor cells is the major limitation to the use of umbilical cord blood (UCB) for the transplantation of adults. We tested the hypothesis that two units transplanted simultaneously could each contribute to haematopoietic reconstitution. A patient with advanced acute lymphocytic leukaemia received a mismatched, unrelated UCB transplant using units from two donors after conditioning. The recipient achieved a complete remission without graft‐versus‐host disease. Double chimaerism was documented in several leucocyte subpopulations; both units contributed to haematopoiesis until relapse. Triple chimaerism was present from relapse until death due to leukaemia. This approach may potentially improve UCB transplantation outcome for adults lacking a histocompatible donor.
An association between human leucocyte antigen alleles and acute and chronic graft‐versus‐host disease after allogeneic haematopoietic stem cell transplantation
Authors: Remberger Mats , Persson Ulla , Hauzenberger Dan , Ringdén Olle
Published: March 2003

Summary. The association between various human leucocyte antigen (HLA) alleles and the occurrence of acute and chronic graft‐versus‐host disease (GVHD) was evaluated in 493 haematopoietic stem‐cell transplant (HSCT) patients with HLA identical sibling donors. There were 307 men and 186 women with a median age of 30 years (0·2–77). Most of the patients had a haematological malignancy and received total body irradiation or busulphan combined with cyclophosphamide as conditioning before transplantation. GVHD prophylaxis consisted of monotherapy with methotrexate (MTX) or cyclosporin (CsA) in 118 patients, MTX + CsA in 323, T‐cell depletion in 28 and other combinations in 24. In total, 84 patients (17%) received a peripheral blood stem‐cell graft, whereas the rest received bone marrow. The cumulative incidence of acute GVHD grades II–IV was 20%, and chronic GVHD 46%. In the multivariate analysis, HLA‐A10 (OR 2·14, CI 1·04–4·41, P = 0·03) and HLA‐B7 (OR 1·80, CI 1·04–3·12, P = 0·03) correlated with an increased risk of acute GVHD grades II‐IV. We also found an association between HLA‐B27 (RR 0·60, CI 0·37–0·95, P = 0·04) and a lower incidence of chronic GVHD. These HLA alleles were independent of other known risk factors for acute or chronic GVHD, as shown by multivariate analysis. These results show that major histocompatibility comlex (MHC) alleles may influence the incidence of GVHD in HSCT with HLA identical sibling donors.
The TNFd4 allele is correlated to moderate‐to‐severe acute graft‐versus‐host disease after allogeneic stem cell transplantation
Authors: Nordlander Anna , Uzunel Mehmet , Mattsson Jonas , Remberger Mats
Published: March 2003

Summary. Certain cytokine gene polymorphisms may be associated with severe acute graft‐versus‐host disease (GVHD) after allogeneic stem cell transplantation. The present study analysed 196 patients and their donors for TNF−308, TNFd, IL‐10−1064 and IL‐10−1082 gene polymorphisms. Serum analysis of tumour necrosis factor‐α (TNF‐α) and interleukin 10 (IL‐10) levels during conditioning therapy was also performed. Among patients with sibling donors, the TNFd allele 4 was significantly correlated with acute GVHD grades II–IV (P < 0·01). Acute GVHD grades II–IV were more common among patients homozygous for the IL‐10−1064 allele 13 (P = 0·02). Patients homozygous for the TNF−308 allele (AA) correlated with higher TNF‐α serum levels during conditioning (P = 0·02).
FLT3 internal tandem duplication in childhood acute myeloid leukaemia: association with hyperleucocytosis in acute promyelocytic leukaemia
Authors: Arrigoni Paolo , Beretta Chiara , Silvestri Daniela , Rossi Vincenzo , Rizzari Carmelo , Valsecchi Maria Grazia , Cazzaniga Giovanni , Biondi Andrea
Published: March 2003

Summary. We evaluated the incidence of FLT3/internal tandem duplication (ITD) mutation in childhood acute myeloid leukaemia (AML) diagnosed over 15 years. FLT3/ITD was found in 10 of 45 (22·2%) non‐acute promyelocytic leukaemia (non‐APL) patients. The 5‐year event‐free survival of non‐APL patients was higher in FLT3/ITD‐negative versus ‐positive patients (48·9%, SE 8·9, vs 20·0%, SE 16·1, P = 0·03). In childhood APL, FLT3/ITD incidence was higher than in non‐APL, although not statistically significant (10 out of 29 patients, 34·5%, P = 0·29). In APL patients, FLT3/ITD was strongly correlated to a higher white blood cell count at diagnosis and the M3 French–American–British subtype.
A novel fusion variant of the MORF and CBP genes detected in therapy‐related myelodysplastic syndrome with t(10;16)(q22;p13)
Authors: Kojima Kensuke , Kaneda Kinuyo , Yoshida Chikamasa , Dansako Hirokata , Fujii Nobuharu , Yano Tomofumi , Shinagawa Katsuji , Yasukawa Masaki , Fujita Shigeru , Tanimoto Mitsune
Published: March 2003

Summary. We report a case of therapy‐related myelodysplastic syndrome (t‐MDS) with t(10;16)(q22;p13), in which novel fusion transcripts of the MORF and CBP genes were detected. In one MORF–CBP fusion transcript, exon 15 of the MORF gene was fused in frame with exon 5 of the CBP gene. In a reciprocal CBP–MORF transcript, exon 4 of the CBP gene was fused in frame with exon 16 of the MORF gene. This is the first reported case of t‐MDS associated with t(10;16), and provides molecular evidence that the novel MORF–CBP and/or CBP–MORF fusion protein(s) might play an important role in the development of t‐MDS.
A possible role for the loss of CD27–CD70 interaction in myelomagenesis
Authors: Katayama Yuta , Sakai Akira , Oue Naohide , Asaoku Hideki , Otsuki Takemi , Shiomomura Takeshi , Masuda Rie , Hino Norihiko , Takimoto Yasuo , Imanaka Fumio , Yasui Wataru , Kimura Akiro
Published: March 2003

Summary. CD27 is a marker of memory B cells and its interaction with its ligand, CD70, is very important for differentiation into plasma cells. Although CD27 is detected on normal plasma cells, its expression is significantly reduced with the progression of multiple myeloma (MM), including monoclonal gammopathy of undetermined significance (MGUS). CD27+ myeloma cells are thought to represent an early phase of myeloma, as CD27+ plasma cells from MM patients were found to be composed of normal plasma cells (CD19+/CD38++) and myeloma cells (CD19–/CD38++), and monoclonality was detected in the CD27+/CD38++ fraction. Given that the lack of CD27 on plasma cells is related to myelomagenesis and that the pro‐apoptotic protein Siva is thought to bind to the cytoplasmic tail of CD27, we analysed alterations of cell growth and genes caused by co‐culturing CD27‐transfected myeloma cell lines (U266, KMS‐5) with CD70‐transfected NIH3T3 cells. CD27–CD70 interaction could not induce apoptosis in either type of myeloma transfectant, and binding between Siva and CD27 was not detected. cDNA microarray (human apoptosis CHIP) analysis showed a significant upregulation of expression of the ectodermal neural cortex 1 (ENC1) gene by CD27–CD70 interaction compared with CD27 transfection alone. These findings show that the relationship between the loss of CD27 and oncogenesis of plasma cells is not simple. It remains unclear whether the lack of CD27 leads to evasion of apoptosis.
Circadian variations of transferrin saturation levels in iron‐overloaded patients: implications for the screening of C282Y‐linked haemochromatosis
Authors: Guillygomarc'h Anne , Christian Jacquelinet , Romain Moirand , Vincent Quentin , Véronique David , Deugnier Yves
Published: March 2003

Summary.The phenotypic screening for genetic haemochromatosis (GH) relies upon the determination of transferrin saturation (TS). In large‐scale screening programs, the time of blood sampling can be uneasy to control. We studied the circadian variations of TS at 08·00 hours, 12·00 hours, 18·00 hours and 00·00 hours in 46 C282Y homozygous patients (GH) and 47 non‐GH patients (NH), to determine whether the time of blood sampling influenced the results of screening. In both groups, there were significant circadian variations in TS, with the highest values at 08·00 hours and the lowest at 00·00 hours. For any given time‐point, TS was significantly higher in the GH group when compared with the NH group (P < 0·0001). For both groups, there was a significant decrease in TS between 08·00 hours and 00·00 hours (P < 0·0001) but this decrease was not as significant in GH when compared with NH patients (interactionP < 0·0073). Receiver operating characteristics (ROC) curves generated for TS at 08·00 hours, 12·00 hours, 18·00 hours and 00·00 hours, presented the same efficiency of diagnosis of GH, with TS threshold varying between 64% at 08·00 hours and 36% at 00·00 hours. In conclusion, for screening studies of C282Y homozygosity, determination of transferrin saturation may be performed at any time during the day.
Common clonal T‐cell origin in a patient with T‐prolymphocytic leukaemia and associated cutaneous T‐cell lymphomas
Authors: Assaf Chalid , Hummel Michael , Dippel Edgar , Schwartz Stefan , Geilen Christoph C. , Harder Lana , Siebert Reiner , Steinhoff Matthias , Klemke Claus‐Detlev , Thiel Eckhard , Goerdt Sergij , Stein Harald , Orfanos Constantin E.
Published: March 2003

Summary. An unusual course was observed in a patient with indolent T‐prolymphocytic leukaemia (T‐PLL) who subsequently developed mycosis fungoides (Mf), lymphomatoid papulosis (LyP) and cutaneous CD30+ anaplastic large cell lymphoma (ALCL). Polymerase chain reaction analysis demonstrated identical monoclonal T‐cell receptor‐β and ‐γ gene rearrangements in all the different clinical entities. Furthermore, cytogenetic studies revealed the same aberrant clone with trisomy of chromosome 8 in T‐PLL and ALCL cells. This unique observation suggests that in T‐PLL, the leukaemic cells might undergo secondary transformation, subsequently resulting in different phenotypes of cutaneous T‐cell lymphoma.
Overexpression of manganese superoxide dismutase promotes survival in cell lines after doxorubicin treatment
Authors: Suresh Alaparthy , Guedez Liliana , Moreb Jan , Zucali James
Published: March 2003

Summary. Overexpression of manganese superoxide dismutase (MnSOD) has been postulated as one possible mechanism of protection from oxidative damage and free radicals. Doxorubicin treatment induces oxygen free radicals, leading to cytotoxicity and myelosuppression. The present study was performed to determine whether over‐expression of MnSOD may play a role in resistance to doxorubicin. Retroviral constructs having the human MNSOD gene in the sense orientation and the neomycin phosphotransferase gene (NEOR) as a selectable marker were transduced into the human melanoma cell line A375 and the human histiocytic lymphoma cell line U937. Stably transduced A375 and U937 cells were subjected to 10–100 ng/ml doxorubicin for 24 h and compared with doxorubicin‐treated A375 and U937 cells transduced with vector only. A colony forming assay was used to determine cell viability in semi‐solid medium. Results demonstrated that wild‐type A375 and U937 cells display low levels of endogenous MnSOD mRNA and protein, and are sensitive to doxorubicin treatment. In contrast, A375 and U937 cells transduced with the MNSOD gene consistently demonstrate increased colony formation in the presence of increasing concentrations of doxorubicin. MnSOD‐transduced A375 and U937 cells also demonstrate increased MnSOD mRNA and protein levels when compared with wild type or those cells transduced with vector only. These results indicate that overexpression of MnSOD can enhance resistance to doxorubicin treatment.
Abnormal expression of apoptosis‐related genes in haematological malignancies: overexpression of MYC is poor prognostic sign in mantle cell lymphoma
Authors: Nagy Bálint , Lundán Tuija , Larramendy Marcelo L. , Aalto Yan , Zhu Ying , Niini Tarja , Edgren Henrik , Ferrer Anna , Vilpo Juhani , Elonen Erkki , Vettenranta Kim , Franssila Kaarle , Knuutila Sakari
Published: March 2003

Summary. The expression of apoptosis‐related genes BCL2, BAX, BCL2L1, BCL2A1, MCL1, DAPK1 and MYC was studied by quantitative real‐time polymerase chain reaction on total RNA samples from patients with acute lymphoblastic leukaemia (ALL, n = 16), acute myeloid leukaemia (AML, n = 27), chronic myeloid leukaemia (CML, n = 12), mantle cell lymphoma (MCL, n = 19) and chronic lymphoid leukaemia (CLL, n = 32). BCL2, BAX, BCL2A1, MCL1, DAPK1 and MYC were overexpressed in all patient groups. BCL2L1 was underexpressed in CLL and CML, but not in AML, ALL and MCL. MCL1 levels were significantly higher in CD13 and CD33‐positive ALL, and in CD56‐positive AML samples. BCL2, BCL2L1, BCL2A1 and MCL1 were overexpressed and DAPK1 was underexpressed in CLL samples with a 11q23 deletion. MYC overexpression was significantly associated with shorter overall survival in MCL (P < 0·01). AML patients with a normal karyotype showed a higher frequency of BCL2A1 overexpression (P < 0·001) than those with an abnormal karyotype.
Changes of isoagglutinin titres after ABO‐incompatible allogeneic stem cell transplantation
Authors: Lee Je‐Hwan , Lee Jung‐Hee , Choi Seong‐Jun , Kim Shin , Seol Miee , Kwon Seog‐Woon , Park Chan‐Jeoung , Chi Hyun‐Sook , Lee Jung‐Shin , Kim Woo‐Kun , Lee Kyoo‐Hyung
Published: March 2003

Summary. We investigated the changes in isoagglutinin titres in 62 patients who underwent ABO‐incompatible allogeneic stem cell transplantation. After major [and/or (±) minor] ABO‐incompatible transplantation, recipient‐derived isoagglutinins against donor‐type red blood cells (RBCs) disappeared more rapidly in unrelated recipients (P = 0·006) and in patients with acute graft‐versus‐host disease (GVHD, P = 0·025) than in sibling recipients and in patients without acute GVHD respectively. Pure red cell aplasia (PRCA) developed in 10 out of 35 evaluable patients who underwent major (± minor) ABO‐incompatible transplantation, and the post‐transplant increase of isoagglutinin titres was a significant predictor for the occurrence of PRCA. In five out of 36 patients who underwent minor (and/or (±) major) ABO‐incompatible transplantation, donor‐derived isoagglutinins against recipient RBCs were detectable without clinically overt haemolysis. Isoagglutinin titres against ABO antigens absent both on recipient and donor RBCs decreased during the early post‐transplant period then rose subsequently in 24 out of 29 patients at (median) d 59 post transplant. Our study showed that changes in isoagglutinin titres might have clinical implications in the occurrence of immunohaematological complications such as PRCA or immune‐mediated haemolysis, and might reflect immunohaematological reconstitution after transplantation. Furthermore, our data regarding time to disappearance of recipient‐derived isoagglutinins against donor‐type RBCs after major ABO‐incompatible transplantation suggest the presence of a graft‐versus‐plasma cell effect.
ADAMTS13 gene defects in two brothers with constitutional thrombotic thrombocytopenic purpura and normalization of von Willebrand factor‐cleaving protease activity by recombinant human ADAMTS13
Authors: Antoine Gerhard , Zimmermann Klaus , Plaimauer Barbara , Grillowitzer Monika , Studt Jan‐Dirk , Lämmle Bernhard , Scheiflinger Friedrich
Published: March 2003

Summary. Genetic analysis of the ADAMTS13 locus identified six mutations in the ADAMTS13 genes of two brothers suffering from constitutional thrombotic thrombocytopenic purpura (TTP): a stop codon leading to a truncated protein on the paternal ADAMTS13 allele and five amino acid exchanges on the maternal allele, three of which were single nucleotide polymorphisms. The other two mutations, not detected in 230 sequenced alleles of healthy control subjects, are, therefore, probably responsible, alone or as part of a combination, for the severe ADAMTS13 deficiency. We also investigated the feasibility of using recombinant ADAMTS13 (rADAMTS13) for normalization of von Willebrand factor‐cleaving protease (VWF‐cp) activity in plasma of the two congenitally deficient patients. Addition of rADAMTS13 to their plasma restored the VWF‐processing pattern to normal, suggesting the potential usefulness of rADAMTS13 for therapy and prophylaxis of familial TTP.
Haematological effects of the C282Y HFE mutation in homozygous and heterozygous states among subjects of northern and southern European ancestry
Authors: Beutler Ernest , Felitti Vincent , Gelbart Terri , Waalen Jill
Published: March 2003

Summary. High frequencies of the C282Y and H63D mutations of the HFE gene occur in European populations, even though homozygous and compound heterozygous states are associated with hereditary haemochromatosis, which is a disease that decreases fitness. This suggests that heterozygotes may possess a selective advantage. HFE mutations increase iron absorption in patients with haemochromatosis, and the mean transferrin saturations and ferritin levels are mildly increased in heterozygotes, suggesting that HFE mutations may protect against iron depletion and iron deficiency anaemia. In this study of 23 681 Caucasian adults, mean transferrin saturation, serum ferritin and haemoglobin levels were significantly higher in subjects carrying HFE mutations compared with wild types. Analysed by ethnicity, mean haemoglobin and mean erythrocyte volume (MCV) were significantly lower in those with a southern versus northern European ancestry. C282Y mutation carriers had an increased mean haemoglobin level in both ethnic groups. Prevalence of non‐anaemic iron deficiency was significantly lower among female carriers of the C282Y mutation compared with HFE wild types. However, prevalence of frank iron deficiency anaemia did not differ significantly among genotypes. Quantile:quantile plots showed a small but significant upward shift in the mid‐range of the haemoglobin distribution among C282Y mutation carriers that was consistent with an increased mean haemoglobin level without significant changes in the anaemic range.