bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic impact of karyotypic findings in childhood acute lymphoblastic leukaemia: a Nordic series comparing two treatment periods
Authors: Forestier Erik , Johansson Bertil , Gustafsson Göran , Borgström Georg , Kerndrup Gitte , Johannsson Johann , Heim Sverre ,   for the Nordic Society of Paediatric Haematology and Oncology (NOPHO) Leukaemia Cytogenetic Study Group
Published: March 2003

The prognostic impact of acquired chromosome abnormalities was evaluated in a population‐based consecutive series of 768 children (< 15 years of age) with acute lymphoblastic leukaemia (ALL). The study cohort included all cases of cytogenetically abnormal childhood ALL diagnosed between 1986 and 1997 in the five Nordic countries (Denmark, Finland, Iceland, Norway and Sweden). The probability of event‐free survival (pEFS) for the total cohort was 0·72 ± 0·02. When comparing the two treatment periods of July 1986 to December 1991 and January 1992 to December 1997, a better survival was seen for the latter time period (pEFS of 0·69 ± 0·02 vs. 0·76 ± 0·02, P = 0·05). Hypodiploidy with less than 45 chromosomes, t(9;22)(q34;q11) and 11q23 translocations were associated with a dismal outcome during the whole study period (pEFS of 0·57 ± 0·12, 0·41 ± 0·14 and 0·37 ± 0·10 respectively). The poor prognostic influence of 11q23 rearrangements seemed to be restricted to infants and older children (> 10 years), who differed significantly from children aged 1–10 years in this regard (P < 0·01). Patients with t(9;22)‐positive ALL seemed to benefit from allogeneic bone marrow transplantation in first remission (P = 0·05). The pEFS for children with t(1;19)(q23;p13)‐positive ALL was intermediate (0·63 ± 0·17), with a tendency to a better outcome for patients with the unbalanced variant der(19)t(1;19). Hyperdiploid ALL patients, subdivided into moderate hyperdiploidy (47–51 chromosomes), massive hyperdiploidy (52–60 chromosomes) and cases in the tri‐/tetraploid range (> 60 chromosomes) had the best outcome in the last treatment period (pEFS of 0·81 ± 0·06, 0·80 ± 0·04 and 0·88 ± 0·07 respectively), unless t(1;19), t(8;14), t(9;22) or 11q23 translocations were present. In a multivariate analysis including white blood cell (WBC) count, immunophenotype, age, mediastinal mass, central nervous system involvement and leukaemia karyotype, only WBC and modal chromosome number were shown to be significant independent risk factors (P < 0·01).
Different levels of prion protein (PrPc) expression on hamster, mouse and human blood cells
Authors: Holada Karel , Vostal Jaroslav G.
Published: March 2003

The host prion protein, PrPc, and its conformationally changed isoform, PrPsc, play an essential role in the transmissible spongiform encephalopathy (TSE) infections. The prion hypothesis postulates that PrPsc is the TSE infectious agent and that it serves as a template to convert host PrPc to additional PrPsc. Blood of experimentally TSE‐infected rodents has been shown to contain TSE infectivity. If blood‐borne TSE infectivity requires association with PrPc, differences in the distribution of PrPc in blood could affect the amount and distribution of blood‐borne infectivity in different hosts. We have compared the distribution of PrPc on the peripheral blood cells of humans, hamsters and mice using quantitative flow cytometry. Human lymphocytes, monocytes and platelets displayed much greater quantities of PrPc than corresponding mouse cells. Mouse platelets did not express any detectable PrPc. A similar low level of PrPc was found on both human and mouse red blood cells. None of the hamster peripheral blood cells displayed detectable amounts of PrPc. If PrPc contributes to the propagation or transport of TSE infectivity in blood, the species differences in PrPc distribution reported here need to be considered when extrapolating the results of rodent TSE transmission studies with blood and blood components to humans.
Effective treatment of Jo‐1‐associated polymyositis with T‐cell‐depleted autologous peripheral blood stem cell transplantation
Authors: Baron Frédéric , Ribbens Clio , Kaye Olivier , Fillet Georges , Malaise Michel , Beguin Yves
Published: March 2003

A patient with Jo‐1 antibody‐associated polymyositis (Jo‐1 PM) had a Karnofsky score of 40% and severe muscle, liver and lung damage that was refractory to standard therapy. The female patient received an autologous T‐cell‐depleted haematopoietic stem cell transplant (HSCT) after myeloablative conditioning. The transplant procedure was complicated by severe adult respiratory distress syndrome (ARDS) and adenovirus‐associated haemorrhagic cystitis as well as cytomegalovirus (CMV) reactivation. The patient's creatinine phosphokinase (CPK) and alanine transaminase (ALT) values were normal on day 21. The patient's strength has improved remarkably and her dyspnoea is subjectively improved. At 15 months after the transplant, the patient was well with a Karnofsky score of 80% and had been off any therapy, including steroids, for 14 months.
Kell is not restricted to the erythropoietic lineage but is also expressed on myeloid progenitor cells
Authors: Wagner T. , Berer A. , Lanzer G. , Geissler K.
Published: March 2003

Anti‐Kell antibodies have been shown to suppress fetal erythropoiesis, but little is known about their effect on myelopoiesis. We analysed the effect of Kell‐related antibodies on granulocyte–macrophage colony‐forming units (CFU‐GM) growth in semisolid medium using peripheral blood mononuclear cells (PBMNCs) from haematologically normal individuals. In addition to its inhibitory effect on erythroid burst‐forming units (BFU‐E) growth, anti‐Kell antibodies significantly reduced CFU‐GM colony formation from Kell‐positive individuals but not from Kell‐negative donors. Moreover, anti‐cellano and anti‐Kpb antibodies also inhibited the growth of CFU‐GM from antigen‐positive MNCs. These data indicate that Kell is not restricted to erythroid blood cells, but is also expressed on myeloid progenitor cells.
An unusual form of persistent polyclonal B lymphocytosis in an infant
Authors: Gomez Pedro , Matutes Estella , Sánchez Joaquin , Garcia Jose M. , Román Jose , Gruszka‐Westwood A. , Torres Antonio
Published: March 2003

We report a case of persistent polyclonal lymphocytosis in an infant. The circulating lymphocytes were of a small to medium size and a small proportion were larger and had lymphoplasmacytoid features. The presence of either an infectious or mutagenic agent was excluded. The polyclonal B‐cell nature of the lymphocyte was demonstrated by immunological markers and confirmed by Southern blot analysis and by polymerase chain reaction targeting immunoglobulin genes. In contrast to the common form of polyclonal lymphocytosis, this case was not associated with HLA‐DR7 and/or abnormalities of chromosome 3, p53 or Bcl2/IgH. Whether this lymphocytosis represents a premalignant or a benign condition remains uncertain, although there has been no progression to date.
Factor V Leiden: the venous thrombotic risk in thrombophilic families
Authors: Lensen Ruud , Rosendaal Frits , Vandenbroucke And Jan , Bertina Rogier
Published: March 2003

Factor V Leiden (FVL) leads to a sevenfold increased risk of venous thrombosis and is present in 50% of individuals from families referred because of unexplained familial thrombophilia. We assessed the association of FVL with venous thromboembolism (VTE) in 12 thrombophilic families of symptomatic probands with FVL in a retrospective follow‐up study. We screened 182 first‐ and second‐degree relatives of the 12 unrelated propositi for the FVL mutation and the occurrence of VTE. The incidence rate of VTE in carriers of FVL (0·56%/year) was about six times the incidence for the Dutch population (0·1%/year). The incidence rate in non‐carriers also appeared to be higher (0·15% per year). At the age of 50 years, the probability of not being affected by VTE was reduced to 75% for carriers and to 93% for non‐carriers (P = 0·009). Identification of carriers of FV Leiden may be worthwhile in young symptomatic individuals and their relatives with a strong positive family history of venous thromboembolism or a history of recurrent venous thrombosis who may be at risk (e.g. pregnancy, use of oral contraceptives). After adjustment for prothrombin G20210A (present in two families), even higher thrombotic incidence rates were found in carriers and non‐carriers of FVL. This makes the presence of other unknown prothrombotic risk factors more probable in these families.
Reduced factor XII levels in patients with the antiphospholipid syndrome are associated with antibodies to factor XII
Authors: Jones D. W. , Gallimore M. J. , Mackie I. J. , Harris S. L. , Winter M.
Published: March 2003

Antibodies to factor XII (FXII) have previously been identified in some patients who were lupus anti‐coagulant‐positive. The relationship between these antibodies and FXII levels appeared to be variable. The aim of the present study was to confirm the presence of antibodies to FXII in patients with well characterized antiphospholipid syndrome (APS) and to establish their potential effect on levels of FXII. Forty‐two patients with APS were studied; 21 patients were found to have either immunoglobulin (Ig)G or IgM antibodies to FXII by enzyme‐linked immunosorbent assay (ELISA) using a highly purified preparation of FXII (> 99% pure). Levels of FXII were statistically significantly lower (P = 0·02) in patients with antibodies to FXII when compared with patients without antibodies to FXII (median = 91 μ/dl, s.d. = 39·1, median = 122 μ/dl, s.d. = 41·1 respectively). Four of the 21 patients with antibodies to FXII were found to have FXII levels below the laboratory normal range. Antibodies to FXII are present in significant numbers of patients with APS and may lead to acquired FXII deficiency.
Lineage‐specific regulation of cell cycle control gene expression during haematopoietic cell differentiation
Authors: Furukawa Yusuke , Kikuchi Jiro , Nakamura Mitsuru , Iwase Satsuki , Yamada Hisashi , Matsuda Michio
Published: March 2003

To maintain the fidelity and integrity of blood formation, the cell cycle is under strict regulation during haematopoietic cell differentiation. To elucidate the molecular mechanisms of cell cycle regulation during haematopoiesis, we examined cell cycle control gene expression during lineage‐specific differentiation from CD34+ progenitor cells. Expression of cyclin‐dependent kinases (cdks) and cyclins, except cdk4, was generally suppressed in CD34+ cells freshly isolated from the bone marrow of healthy volunteers. Among four major cdk inhibitors, p16 was expressed more highly in CD34+ cells than in CD34‐negative bone marrow mononuclear cells, whereas the amounts of p21 and p27 transcripts increased in the CD34− population. The behaviour of cell cycle control genes during haematopoietic differentiation was classified into four patterns: (i) universal upregulation (cdc2, cdk2, cyclin A, cyclin B and p21); (ii) upregulation in specific lineages (cyclin D1, cyclin D3 and p15); (iii) no induction or stable expression (cdk4, cyclin D2, cyclin E and p27); and (iv) universal downregulation (p16). Lineage‐specific changes included the sustained elevation of cdc2 and cyclin A during erythroid differentiation, cyclin D1 and p15 induction in myeloid lineage and selective upregulation of cyclin D3 in megakaryocytes. Blocking induction of cyclin D3 resulted in the inhibition of megakaryocytic differentiation. These results suggest that the expression of cell cycle control genes is distinctively regulated in a lineage‐dependent manner, reflecting the cell cycle characteristics of each lineage. Some of these genes play an essential role in the process of differentiation itself.
Retrovirus‐mediated transfer of the herpes simplex virus thymidine kinase and enhanced green fluorescence protein genes in primary T lymphocytes
Authors: Di Florio Sabrina , Sebastiani Carla , Fagioli Marta , Di Ianni Mauro , Alfonsi Diego , Venditti Gigliola , Pelicci Pier Giuseppe , Tabilio Antonio
Published: March 2003

The EGFP‐tk retroviral vector, encoding enhanced green fluorescent protein (EGFP) and the herpes simplex virus thymidine kinase (HSV‐tk) packaged in a Phoenix amphotropic cell line, was used to transduce healthy donor T lymphocytes. Infection yielded a mean of 41·8 ± 9·3% SD (range 31·1–48·4%) EGFP‐positive cells and a mean of 92 ± 2% SD (range 90–94%) after cell sorting. EGFP expression remained stable for 30 d after infection. The entire gene transfer procedure had no significant effect on lymphocyte subsets and slightly reduced clonogenicity. Ganciclovir (gcv) treatment (1 µg/ml × 10 d) killed all EGFP‐positive cells in the transduced and transduced/sorted populations, but had no effect on untransduced controls. Our results show that primary T lymphocytes can be transduced using an EGFP‐tk vector that yields a homogeneous infected population without affecting lymphocyte subsets, function and clonogenicity.
Fibrinogen polymorphisms are not associated with the risk of myocardial infarction
Authors: Doggen C. J. M. , Bertina R. M. , Cats V. Manger , Rosendaal F. R.
Published: March 2003

In the Study of Myocardial Infarctions Leiden, we investigated the prevalence of three polymorphisms in the α‐ and β‐fibrinogen genes among 560 patients with a myocardial infarction and 646 control subjects. Secondly, we studied the relationships between these polymorphisms and fibrinogen activity and antigen levels. The TaqI, HaeIII and BclI polymorphisms in the fibrinogen gene were not associated with myocardial infarction. As we found an association of the rare B2 allele with fibrinogen levels and a similar, but weak, effect for the rare H2 allele, we conclude that a genetic propensity to high fibrinogen levels does not affect the risk of myocardial infarction. This is evidence against a causal role for fibrinogen levels in the aetiology of myocardial infarction.
Cytomegalovirus colitis after autologous transplantation for multiple myeloma
Authors: Ramsey D. J. C. , Schey S. A.
Published: March 2003

Bone marrow or stem cell transplantation is an established therapy for haematological malignancies. We report a cytomegalovirus (CMV) IgG +ve 56‐year‐old patient who underwent autologous rescue with CD34+ selected peripheral blood stem cells as part of consolidation therapy for multiple myeloma and subsequently developed CMV colitis. In contrast to infection secondary to human immunodeficiency virus (HIV), CMV colitis has not previously been described in this context. We discuss this case and issues arising from it related to the use of CD34+ selected stem cells for transplantation.
SIHONSCORE: a scoring system for external quality control of leukaemia/lymphoma immunophenotyping measuring all analytical phases of laboratory performance
Authors: Kluin‐Nelemans J. C. , Van Wering E. R. , Van Der Schoot C. E. , Adriaansen H. J. , Van't Veer M. B. , Van Dongen J. J. M. , Gratama J. W. ,   on behalf of the Dutch Co‐operative Study Group on Immunophenotyping of Haematological Malignancies (SIHON)
Published: March 2003

For the diagnosis of leukaemia and leukaemic lymphoma, clinicians frequently have to rely on the results of immunophenotyping. To improve the quality of these results, the Dutch Foundation for Immunophenotyping of Haematological Malignancies (SIHON) initiated external quality rounds in 1986. Over a period of more than 10 years, this has led to improvements in the interpretation of immunophenotyping results. However, the evaluation of results focused mainly on the correctness of the interpretation of the immunophenotypical data, leaving the preceding analytical phases unevaluated. Therefore, in 1996 SIHON developed a more comprehensive scoring system, called SIHONSCORE, covering all three phases of immunophenotyping, namely the pre‐analytical (i.e. choice of the staining panels), analytical (i.e. the technical part consisting of sample preparation, data acquisition and analysis) and the post‐analytical phase (i.e. the interpretation) of the laboratory process. Here, we report how SIHONSCORE was successfully applied to three consecutive external quality rounds consisting of a total of nine different cases tested. For laboratory certification, participation in external quality control programmes is required. Evidently, criteria are needed to define the minimum acceptable performance of a certified laboratory. With SIHONSCORE, a useful instrument is obtained evaluating all phases of the performance of laboratories in leukaemia and lymphoma immunophenotyping.
Long‐term expansion and maintenance of cord blood haematopoietic stem cells using thrombopoietin, Flt3‐ligand, interleukin (IL)‐6 and IL‐11 in a serum‐free and stroma‐free culture system
Authors: Lazzari Lorenza , Lucchi Simona , Rebulla Paolo , Porretti Laura , Puglisi Giuseppe , Lecchi Lucilla , Sirchia Girolamo
Published: March 2003

Although cord blood (CB) compares favourably with other haematopoietic stem cell (HSCs) sources, its use in large patients is limited by the low number of cells available. Ex vivo expansion of CB HSCs has been used to overcome this limitation. In this study, we investigated the effect of different cytokine cocktails, including interleukin (IL)‐6, IL‐11, Flt3‐ligand (FL) and thrombopoietin (TPO) combined with serum or serum‐free medium on the ex vivo expansion of CD34+ cells from CB. Initial experiments showed that expansion could be slightly improved using serum, but we chose to use serum‐free medium in the subsequent investigations to apply good medical practice (GMP) conditions suitable for clinical use. The highest expansion of CD34+ cells was obtained with a cocktail containing FL + TPO + IL‐6 + IL‐11. The median (range) fold expansions of CD34+ cells at 5 and 10 weeks with serum‐free medium were 235·6 (131·3–340) and 5205·6 (4736·6–5674·7) respectively. The absence of IL‐11 was associated with a similar fold expansion after 5 weeks (median 215·6, range 149·8–281·5), but after 10 weeks expansion was slightly lower (median 1314·7, range 645–1984·4). Our data support the possibility of maintaining long‐term expansion of CB HSCs in a simple stroma‐ and serum‐free system.
The impact of attaining a minimal disease state after high‐dose melphalan and autologous transplantation for multiple myeloma
Authors: Davies Faith E. , Forsyth Peter D. , Rawstron Andrew C. , Owen Roger G. , Pratt Guy , Evans Paul A. S. , Richards Stephen J. , Drayson Mark , Smith Graeme M. , Selby Peter J. , Child J. Anthony , Morgan Gareth J.
Published: March 2003

Initial studies with high‐dose therapy (HDT) in myeloma suggest some beneficial effects of attaining a complete response (CR); however, the effect on survival is difficult to assess owing to inconsistencies in the definition of response between studies. We have analysed 96 newly diagnosed patients aged less than 65 years who received HDT and assessed the effect of response on survival using electrophoresis, immunofixation and fluorescent IgH polymerase chain reaction (PCR) to define CR. Patients received induction chemotherapy with C‐VAMP (adriamycin, vincristine, methylprednisolone, cyclophosphamide) followed by melphalan 200 mg/m2 and reinfusion of peripheral blood stem cells. There was a high response to C‐VAMP [CR = 24%, partial response (PR) = 64%], with all but one patient improving the depth of response after HDT (CR = 69%, PR = 31%). The progression‐free survival (PFS) and overall survival (OS) were excellent at a median of 46·4 months and 72+ months. There was a trend towards an improved PFS in patients with an immunofixation‐negative CR compared with patients with a PR (49·4 months, 41·14 months; P = 0·26). This was not evident when electrophoresis was used to define CR. The method used to define CR did not impact on the overall survival and fluorescent IgH PCR failed to add any additional prognostic information. This study supports the widespread use of the European Bone Marrow Transplantation group (EBMT) response criteria and suggests that immunofixation should be performed on all patients who become electrophoresis negative.
Progressive reduction in treatment‐related deaths in Medical Research Council childhood lymphoblastic leukaemia trials from 1980 to 1997 (UKALL VIII, X and XI)
Authors: Hargrave D. R. , Hann I. M. , Richards S. M. , Hill F. G. , Lilleyman J. S. , Kinsey S. , Bailey C. C. , Chessells J. M. , Mitchell C. , Eden O. B. ,   on behalf of the Medical Research Council Working Party for Childhood Leukaemia
Published: March 2003

In the last 20 years, the survival rate for children with acute lymphoblastic leukaemia (ALL) has markedly improved, largely owing to a decrease in relapses. However, children still die from complications of treatment and these are potentially preventable. We have analysed data from three large consecutive national protocols for ALL from 1980 to 1997 [Medical Research Council United Kingdom ALL (MRC UKALL) trials VIII, X and XI] to compare the incidence and causes of treatment‐related deaths (TRD). The percentage of TRD has fallen from 9% to 2% (UKALL VIII to XI), largely as a result of a decrease in fatal infections. Deaths during induction have fallen from 3% to 1%, the main causes of death being bacterial, followed by fungal infection, while other causes, chiefly haemorrhage, have not declined. Remission deaths also decreased from 6% to 1%, particularly those deaths due to measles and pneumocystis carinii. More guidelines for surveillance and treatment of infections have been included within progressively more intensive protocols. Risk factor analysis showed increased TRD in patients with Down's syndrome, high leucocyte count and older age in UKALL XI. In contrast, the introduction of blocks of intensification was not associated with an increased death rate. While improved supportive care has reduced the incidence of TRD, there is still scope for further reduction by prompt treatment of suspected infection. Maintenance of herd immunity remains of vital importance in avoiding deaths from measles.
Cord blood transplantation from HLA‐mismatched unrelated donors as a treatment for children with haematological malignancies
Authors: Ohnuma Kei , Isoyama Keiichi , Ikuta Koichiro , Toyoda Yasunori , Nakamura Junko , Nakajima Fumiaki , Tsuchida Masahiro , Ohira Mutsuro , Suminoe Aiko , Hara Toshiro , Nishihira Hirokazu
Published: March 2003

Factors influencing the outcome for 39 children with haematological malignancy who were subjected to a cord blood transplantation (CBT) from genotypically HLA‐mismatched unrelated donors were analysed. This retrospective study included 21 children with acute lymphoblastic leukaemia, 15 with acute myelogenous leukaemia and one each with chronic myelogenous leukaemia, refractory anaemia with myelodysplastic syndrome (MDS) and juvenile myelomonocytic leukaemia (JMML). Those subjected to CBT during the first or second complete remission (CR) and MDS without blasts were assigned to the standard‐risk (SR) group (n = 16). Patients in third or subsequent remission, relapse or partial remission with refractory leukaemia at the time of CBT were considered to be in advanced phase, and placed in the high‐risk (HR) group (n = 11). JMML and the second CR after a relapse (n = 8), or bone marrow failure after a rejection (n = 3), following haematopoietic stem cell transplantation (HSCT) in the first CR were included in the high‐risk group. Kaplan–Meier estimates for neutrophil and platelet recovery were 83·7 ± 12·2 at d 60 and 55·4 ± 16·6% at d 100 respectively. The incidence of grades II–VI acute graft‐versus‐host disease was 58·5 ± 16·8%. The Kaplan–Meier estimate for 3‐year event‐free survival (EFS) was 49·2 ± 16·6. From multivariate analysis, the most important factor influencing EFS was disease status at CBT: SR patients had a 3‐year EFS of 75·0 ± 21·6%, compared with 29·6 ± 20·6% for those with HR disease (P = 0·013, RR 4·746, 95% CI 1·382–16·298). These data confirm that HLA‐mismatched, unrelated CBT is a feasible procedure to cure a significant proportion of children with leukaemia, especially if conducted in a favourable phase of the disease.
A dose of 75 μg/kg/d of i.v. anti‐D increases the platelet count more rapidly and for a longer period of time than 50 μg/kg/d in adults with immune thrombocytopenic purpura
Authors: Newman Gregg C. , Novoa Maria V. , Fodero Erin M. , Lesser Martin L. , Woloski B. M. R. , Bussel James B.
Published: March 2003

Treatment with 75 μg/kg/d intravenous (i.v.) anti‐D was compared with 50 μg/kg/d in a prospective randomized study of 27 RhD‐positive, human immunodeficiency virus‐negative, adult, acute, non‐splenectomized patients with immune thrombocytopenic purpura (ITP) and platelet counts ≤ 30 × 109/l. The higher dose resulted in greater median d 1 (43 × 109/l vs. 7·5 × 109/l; P = 0·012) and d 7 (153 × 109/l vs. 64·5 × 109/l; P = 0·001) platelet increases despite no greater haemoglobin decrease. Children with acute ITP receiving 75 μg/kg/d had overnight platelet increases in seven out of nine cases. The duration of effect at the 75 μg/kg/d dose was 46 d vs. 21 d (P = 0·03). Adverse events were mild to moderate and ameliorated with prednisone and acetaminophen premedication.
The Polycomb group protein EZH2 is upregulated in proliferating, cultured human mantle cell lymphoma
Authors: Visser Hein P. J. , Gunster Marco J. , Kluin‐Nelemans Hanneke C. , Manders Erik M. M. , Raaphorst Frank M. , Meijer Chris J. L. M. , Willemze Roel , Otte Arie P.
Published: March 2003

Polycomb group (PcG) proteins are involved in the stable transmittance of the repressive state of their gene targets throughout the cell cycle. Mis‐expression of PcG proteins can lead to proliferative defects and tumorigenesis. There are two separate multimeric PcG protein complexes: an EED–EZH2‐containing complex and a BMI1–RING1‐containing complex. In the normal human follicle mantle, both PcG complexes have mutually exclusive expression patterns. BMI1–RING1 is expressed, but EZH2–EED is not. Here, we studied the expression of both complexes in six cases of mantle cell lymphoma (MCL), which is derived from the follicle mantle. MCL cells can be cultured in vitro and stimulated to proliferation. We found that resting MCL cells expressed BMI1–RING1, but not EZH2–EED, like normal mantle cells. Proliferating MCL cells, however, showed strongly enhanced expression of EZH2. Also, BMI1 and RING1 continued to be expressed in proliferating MCL. This is the first demonstration that EZH2 expression can be upregulated in fresh lymphoma cells. To test whether the enhanced EZH2 expression was causal for the increased proliferation in MCL, we overexpressed EZH2 in two different cell lines. In the B cell‐derived Ramos cell line, EZH2 overexpression caused an increase in the proliferation rate. This suggests a possible causal effect between EZH2 upregulation and increased proliferation in haematopoietic cells.
Health and functional status of adult recipients 1 year after allogeneic haematopoietic stem cell transplantation
Authors: Socié Gérard , Mary Jean‐Yves , Esperou Hélène , Robert Dominique Vexiau , Aractingi Sélim , Ribaud Patricia , Devergie Agnès ,   Marie‐Elisabeth Toubert,, Boudou Patrice , Cathelinau Beryl , Gluckman Eliane , Vexiau Patrick
Published: March 2003

Increasing numbers of patients are surviving after allogeneic haematopoietic stem cell transplantation (SCT). Among these patients, a number of late complications have been described but few data on the risk factors of these long‐term effects of SCT are available. We report the analysis on 105 adult patients, surviving free of haematological disease at a median time of 15 months after SCT. At the time of screening, 52% had returned to work, general health status was normal in 67% and 47% were sexually active. Female patient gender odds ratio (OR) 2·9 (P = 0·01) and age > 25 years (OR = 3·2, P = 0·02) were associated with non‐return to work. Decreased general status was associated with chronic graft‐versus‐host disease (GvHD) (OR = 3·2, P = 0·009) and irradiation (OR = 3·6, P = 0·004). Sexual inactivity was associated with younger age (OR = 7·0, P = 0·0002) and chronic GvHD (OR = 3·3, P = 0·006). Risk factors for altered pulmonary function tests included previous smoking habits, irradiation and chronic GvHD. Obstructive lung disease was associated with a previous history of asthma. Sicca syndrome and conjunctivitis were increased in patients with previous acute GvHD and cataracts were less frequent in patients with aplastic anaemia. Persistent impaired hair re‐growth was less frequent in patients who received irradiation (OR = 0·18, P = 0·002) but increased in patients with previous acute GvHD (OR = 5·3, P = 0·007). Microalbuminuria was more frequent in irradiated patients (OR = 9·4, P = 0·05). Raised cholesterol was associated with age (OR = 20·8, P < 0·001), previous acute GvHD (OR = 4·7, P = 0·03), steroid use (OR = 6·3, P = 0·001) and familial hypercholesterolaemia (OR = 4·4, P = 0·04). Decreased bone density was associated with chronic GvHD (OR = 3·9, P = 0·001). Thus, using routine tests in adult patients we were able to detect significant numbers of‐non‐symptomatic complications enabling early treatment.
Decreased yield, phenotypic expression and function of immature monocyte‐derived dendritic cells in cord blood
Authors: Liu Enmei , Tu Wenwei , Law Helen K. W. , Lau Yu‐Lung
Published: March 2003

Dendritic cells are critical for the induction of both primary immune responses and immunological tolerance, as well as for the regulation of T‐helper 1 (Th1) and 2 (Th2) immune responses. As neonates are notably deficient in Th1 response and cord blood transplantation is noted to result in less graft‐versus‐host disease (GvHD), we compared the phenotypic and functional characteristics of monocyte‐derived dendritic cells (DCs) that favour Th1 development from cord blood and adult peripheral blood to understand the underlying mechanisms of these observations. Our results showed that: (1) after culture for 7 d with interleukin (IL)‐4 and granulocyte–macrophage colony‐stimulating factor (GM‐CSF), cord blood monocytes generated less CD1a+ cells than adult peripheral blood monocytes, and the CD1a+ cell percentage decreased thereafter; (2) compared with adult blood DCs, cord blood DCs had reduced intensity of expression of CD1a and MHC class II molecules, but the expression levels of CD11c and CD86 were similar; (3) the endocytotic ability of cord blood DCs was reduced compared with adult blood DCs, and this function was related to reduced mannose receptor (MR)‐positive cells; (4) furthermore, the ability of cord blood DCs to stimulate CD3+ T cells in an allogeneic mixed lymphocyte reaction was significantly lower than that of adult blood DCs. These results suggested that the dysfunction of cord blood monocytes in differentiating into professional DCs will affect the activation of naive T cells, especially Th1 development, and may be related to the susceptibility to different infections in the neonates, as well as the lower incidence of GvHD in cord blood transplantation.
High‐grade lymphoma mimicking bone crisis in Gaucher's disease
Authors: Manz M. , Riessen R. , Poll L. , Dohmen B. , Kaiserling E. , Krück W. , Laniado M. , Kanz L. , Denzlinger C.
Published: March 2003

A 33‐year‐old woman with type 1 Gaucher's disease developed painful swelling of her right tibia. Initial diagnostics suggested a typical bone crisis. However, clinical course and subsequent imaging pointed to malignant disease, which was specified as high‐grade lymphoma. Chemotherapy was applied together with enzyme replacement and resulted in complete remission of the lymphoma. We conclude that osseous lesions, although suggestive of common manifestations of Gaucher's disease, should be discriminated very carefully from neoplastic infiltration to maintain curative treatment options.
Molecular quantification of viral load in plasma allows for fast and accurate prediction of response to therapy of Epstein–Barr virus‐associated lymphoproliferative disease after allogeneic stem cell transplantation
Authors: Van Esser Joost W. J. , Niesters Hubert G. M. , Thijsen Steven F. T. , Meijer Ellen , Osterhaus Albert D. M. E. , Wolthers Katja C. , Boucher Charles A. B. , Gratama Jan Willem , Budel Leo M. , Van Der Holt Bronno , Van Loon Anton M. , Löwenberg Bob , Verdonck Leo F. , Cornelissen Jan J.
Published: March 2003

Epstein‐Barr virus lymphoproliferative disease (EBV‐LPD) following allogeneic stem cell transplantation (allo‐SCT) has a poor prognosis. We used a sensitive real‐time polymerase chain reaction (PCR) assay for quantitative detection of EBV‐DNA in plasma and serially measured EBV‐DNA levels to assess the response to treatment in allo‐SCT recipients with EBV‐LPD. Fourteen allo‐SCT recipients with EBV‐LPD who received a T cell‐depleted (TCD) sibling (n = 5) or matched unrelated donor (n = 9) graft were monitored from the time of EBV‐LPD diagnosis, during therapy and assessment of clinical response. Seven patients had complete responses of EBV‐LPD to therapy, of whom 21% (3 out of 14) survived beyond 6 months from EBV‐LPD diagnosis. Clinically responding patients showed a rapid decline of EBV‐DNA plasma levels within 72 h from the start of therapy. In contrast, all clinical non‐responders showed an increase of EBV‐DNA levels. Absolute EBV‐DNA levels at the time of EBV‐LPD diagnosis did not predict for response, but the pattern of EBV‐DNA levels within 72 h from the start of therapy (> 50% decrease versus increase) strongly predicted for clinical response (P = 0·001). Quantitative monitoring of EBV‐DNA levels from the start of and during therapy for EBV‐LPD rapidly and accurately predicts for response to therapy as early as within 72 h. It may thus provide a powerful tool to adjust and select treatment in individuals with EBV‐LPD following allo‐SCT.
Cytogenetic findings and their clinical relevance in myelofibrosis with myeloid metaplasia
Authors: Tefferi Ayalew , Mesa Ruben A. , Schroeder Georgene , Hanson Curtis A. , Li Chin‐Yang , Dewald Gordon W.
Published: March 2003

The prognostic significance of bone marrow cytogenetic lesions in myelofibrosis with myeloid metaplasia (MMM) was investigated in a retrospective series of 165 patients. An abnormal karyotype was demonstrated in 57% of patients. At diagnosis (n = 92), 48% of the patients had detectable cytogenetic abnormalities, and clonal evolution was frequently demonstrated in sequential studies. More than 90% of the anomalies were represented by 20q–, 13q–, +8, +9, 12p–, and abnormalities of chromosomes 1 and 7. Of these, 20q–, 13q– and +8 were the most frequent sole abnormalities, each occurring in 15–25% of the abnormal cases. Trisomy 9 and abnormalities of chromosomes 1 and 7 were equally prevalent but were usually associated with additional cytogenetic lesions. Chromosome 5 abnormalities were infrequent but were over‐represented in the group of patients exposed to genotoxic therapy. In a multivariate analysis that incorporated other clinical and laboratory variables, the presence of an abnormal karyotype did not carry an adverse prognosis. Instead, +8, 12p–, advanced age and anaemia were independent prognostic determinants of inferior survival. In particular, survival was not adversely affected by the presence of either 20q– or 13q–.
Acute myeloid leukaemia M0: haematological, immunophenotypic and cytogenetic characteristics and their prognostic significance: an analysis in 241 patients
Authors: Béné Marie‐Christine , Bernier Michel , Casasnovas René Olivier , Castoldi Gianluigi , Doekharan Dew , Van Der Holt Bronno , Knapp Walter , Lemež Petr , Ludwig Wolf‐Dieter , Matutes Estella , Orfao Alberto , Schoch Claudia , Sperling Christian , Van′t Veer Mars B. ,   for the European Group for the Immunological Characterization of Leukaemias (EGIL)
Published: March 2003

Haematological, immunophenotypic and cytogenetic characteristics were analysed in 241 patients with acute myeloid leukaemia (AML) M0, including 58 children. Children < 3 years and adults between 60 and 70 years of age were most frequently affected. Immunophenotyping showed a heterogeneous phenotype. Anti‐myeloperoxidase was positive in about half of the patients. Cytogenetic data were available from 129 (54%) patients. A normal karyotype was found in only 24%. Most of the abnormalities were unbalanced and the chromosomes 5, 7, 8 and 11 were the most frequently affected. Survival data were available from 152 treated patients (63%). The median overall survival for all patients was 10 months, 20 months for children (n = 36), 10 months for the young adult group (n = 50) and 7 months for the elderly patients (n = 66) (P = 0·09). Karyotype was not a prognostic factor influencing survival. AML M0 shows the immunological characteristics of early progenitor cells, but the expression of the different markers and cytogenetic abnormalities is heterogeneous. The prognosis is poor compared with other de novo AML and similar to that of AML with multilineage dysplasia or AML following myelodysplastic syndromes.
Detection of reciprocal fusion 5′‐BCL6/partner‐3′ transcripts in lymphomas exhibiting reciprocal BCL6 translocations
Authors: Kaneita Yoshitaka , Yoshida Shoko , Ishiguro Naoko , Sawada Umihiko , Horie Takashi , Mori Shigeo , Moriyama Masatsugu
Published: March 2003

It has been believed that replacement of the endogenous promoter and the non‐coding first exon of the BCL6 gene by a sequence derived from the translocational partner gene is a main mechanism of the BCL6 dysregulation resulting from translocation. In this study, we found that reciprocal BCL6 translocation led to the expression of not only the 5′‐partner/BCL6–3′ fusion transcripts but also the 5′‐BCL6/partner‐3′ fusion transcripts, suggesting that reciprocal 5′‐BCL6/partner‐3′ fusion genes are transcriptionally active. These findings raise the possibility that reciprocal BCL‐6 translocation may lead to dysregulation of the partner gene as well as the BCL6 gene.
Ten years of prophylactic treatment with fresh‐frozen plasma in a child with chronic relapsing thrombotic thrombocytopenic purpura as a result of a congenital deficiency of von Willebrand factor‐cleaving protease
Authors: Barbot José , Costa Elísio , Guerra Marisol , Barreirinho Maria S. , Isvarlal Pratima , Robles Rodolfo , Gerritsen Helena E. , Lämmle Bernhard , Furlan Miha
Published: March 2003

We report the results of 10 years of prophylactic fresh‐frozen plasma (FFP) infusion therapy in a 14‐year‐old girl with chronic relapsing thrombotic thrombocytopenic purpura (TTP), in whom a severe congenital von Willebrand factor (VWF)‐cleaving protease deficiency has been documented. Severe haemolytic crises triggered by infections were prevented and her present renal and neurological functions have been preserved. Sequential measurements of protease activity and platelet count after FFP infusion led us to conclude tentatively that 5% may be sufficient to degrade very large and adhesive VWF multimers.
Proliferative activity of leukaemic blasts and cytosine arabinoside pharmacodynamics are associated with cytogenetically defined prognostic subgroups in acute myeloid leukaemia
Authors: Braess Jan , Jahns‐Streubel Gerlinde , Schoch Claudia , Haase Detlef , Haferlach Torsten , Fiegl Michael , Voss Stefanie , Kern Wolfgang , Schleyer Eberhardt , Hiddemann Wolfgang
Published: March 2003

The biological mechanisms responsible for the association of specific karyotypes with prognosis in acute myeloid leukaemia (AML) remain largely unclear. A prospective study was performed to evaluate how far cytogenetically defined prognostic subgroups of AML differ in their proliferative activity as a potential mechanism for differential sensitivities to S‐phase‐specific induction chemotherapy comprising cytosine arabinoside (AraC). One hundred and eighty‐seven patients with de novo AML were included in the study; 25 patients with a favourable [inv(16), t(8;21), t(15;17)] karyotype, 99 with a normal karyotype, 29 with an unfavourable karyotype (−5, 5q–, −7, 7q–, complex aberrations) and 34 with cytogenetic aberrations of unknown prognostic significance (all others). The favourable group demonstrated the highest ex vivo proliferative activity (PA) (3·41 pmol/105 cells), significantly (P = 0·02) exceeding the unfavourable group with the lowest PA (0·72) and the group with a normal karyotype (1·06) or with karyotype of unknown significance (1·05) that both demonstrated an intermediate PA. Samples with a high PA (> median of the whole group) were more likely to produce interleukin 3, granulocyte macrophage colony‐stimulating factor (GM‐CSF), granulocyte CSF (G‐CSF) (56%, 43% and 50%) than cells with a low PA (33%, 36% and 36%; n.s.). The effect of priming by exogenous GM‐CSF or G‐CSF was significantly more pronounced in samples with a low PA than in rapidly proliferating samples (P < 0·01). For the whole group, a high PA was closely associated with an increased incorporation of AraC triphosphate (AraCTP) into the DNA (P < 0·0001). Clinically, a high PA was associated with a better complete remission (CR) rate in the normal (95% versus 62%) and the unfavourable group (75% versus 33%). The significant differences in proliferative activity between cytogenetic subgroups of AML are associated with increased cytosine arabinoside pharmacodynamics and constitute one potential mechanism for the different response of cytogenetic subgroups to AraC‐based induction therapy.
Profiling of genes expressed in human monocytes and monocyte‐derived dendritic cells using cDNA expression array
Authors: Lapteva Natalia , Ando Yoshitaka , Nieda Mie , Hohjoh Hirohiko , Okai Miki , Kikuchi Akiko , Dymshits Grigory , Ishikawa Yoshihide , Juji Takeo , Tokunaga Katsushi
Published: March 2003

Using a human cDNA expression array, we obtained expression profiles of 588 genes in CD14+monocytes and monocyte‐derived dendritic cells (DCs). Overall, 22 genes were upregulated, and nine genes were downregulated in DCs of both samples from two different individuals. Many of the genes that were upregulated in DCs encode proteins that are related to differentiation, cell structure, migration, termination of cell cycle as well as proliferation, e.g. tumour necrosis factor‐α (TNF‐α), tumour necrosis factor receptor II (TNFRII), thymosin β‐10, epithelial discoidin domain receptor 1, replication factor C, putative transcription factor DB1, alpha catenin, transforming growth factor‐β1, prohibitin, p53‐regulating protein and neu differentiation factor. Among the downregulated genes in DCs were genes that encode proteins of cell cycle regulation: mitotic growth and transcription activator, platelet‐derived growth factor receptor‐β subunit, interleukin 2 receptor (IL‐2R)‐γ subunit, IL‐7R‐α subunit, leucocyte interferon‐γ (IFN‐γ) and granulocyte–macrophage colony‐stimulating factor receptor (GM‐CSFR). Semi‐quantitative reverse transcription–polymerase chain reaction method confirmed the upregulated expression levels in DCs for TNFRII, TNF‐α, alpha catenin and downregulation of IFN‐γ, GM‐CSFR on four different donor samples of DCs and monocytes. Moreover, our data show the presence of a ‘switch‐on’ step for the TNF‐α and TNFRII gene expression in immature DCs for further differentiation into mature DCs.
Successful treatment with cyclosporin A for myelodysplastic syndrome with erythroid hypoplasia associated with T‐cell receptor gene rearrangements
Authors: Shimamoto Takashi , Iguchi Tomotaka , Ando Keiko , Katagiri Tomoko , Tauchi Tetsuzo , Ito Yoshikazu , Yaguchi Makoto , Miyazawa Keisuke , Kimura Yukihiko , Masuda Michihiko , Mizoguchi Hideaki , Ohyashiki Kazuma
Published: March 2003

Myelodysplastic syndrome (MDS) with erythroid hypoplasia, a rare form of MDS, has not yet been clearly defined. We report four patients with MDS with erythroid hypoplasia who received immunosuppressive therapy. All were elderly, had severe transfusion‐dependent anaemia, morphological evidence of myelodysplasia and a low percentage (3·2–13·6%) of erythroid precursors. Administration of cyclosporin A (CsA) improved their anaemia; all transfusion‐dependent patients achieved transfusion‐independence. An inverted CD4/8 ratio was seen in three patients who also demonstrated T‐cell receptor (TCR)‐β and ‐γ gene rearrangements by Southern blotting and clonality by polymerase chain reaction. Treatment with CsA can be an attractive alternative treatment for patients with MDS with erythroid hypoplasia, which may be associated with a clonal abnormality in T cells.
Early‐acting cytokine‐driven ex vivo expansion of mobilized peripheral blood CD34+ cells generates post‐mitotic offspring with preserved engraftment ability in non‐obese diabetic/severe combined immunodeficient mice
Authors: Herrera C. , Sánchez J. , Torres A. , Bellido C. , Rueda A. , Alvarez M. A.
Published: March 2003

The ability of ex‐vivo expanded peripheral blood stem cells (PBSC) to engraft non‐obese diabetic/severe combined immunodeficient (NOD/SCID) mice has not been evaluated to date. We investigated the maintenance of primitive SCID‐repopulating cells (SRC) and long‐term culture‐initiating cells (LTCIC) in PBSC expanded with early‐acting cytokines, thrombopoietin (TPO), stem cell factor (SCF) and FlT3‐ligand (FL) with or without interleukin 3 (IL‐3) and IL‐6 in short‐term (6 d) stroma‐free serum‐free cultures. TPO + SCF + FL and TPO + SCF + FL + IL‐3 + IL‐6 produced 5·9 ± 1·97 and 18·25 ± 4·49 (mean ± SEM)‐fold increase of CD34+ cells respectively. We tracked cellular division with PKH26 and sorted post‐mitotic CD34+ PKH26low cells to assess their primitive functional properties. After culture with TPO + SCF + FL, LTCICs among post‐mitotic cells increased 12·08 ± 3·4 times, and 4·3 ± 1·6 times when IL‐3 + IL‐6 were added. CD34+ PKH26low cells cultured with TPO + SCF + FL provided human multilineage (CD34, CD33 and CD19) engraftment in NOD/SCID mice, whereas no human cells were detected in mice injected with cells cultured with TPO + SCF + FL + IL‐3 + IL‐6. Percentages of CD34+/CD38, CD34+/CD33, CD34+/DR and cells in G0/G1 phase were similar among cells cultured with both cytokine combinations, indicating that the deleterious impact of IL‐3 + IL‐6 on the ability to engraft is not translated into phenotypic or cycling features. In conclusion, TPO + SCF + FL‐expanded PBSC maintain multilineage engraftment ability in NOD/SCID mice, which is abrogated by the addition of IL‐3 + IL‐6.
Autologous stem cell transplantation in elderly multiple myeloma patients over the age of 70 years
Authors: Badros Ashraf , Barlogie Bart , Siegel Eric , Morris Christopher , Desikan Raman , Zangari Maurizio , Fassas Athanasios , Anaissie Elias , Munshi Nikhil , Tricot Guido
Published: March 2003

The feasibility and efficacy of autologous stem cell transplantation (auto‐SCT) in patients aged ≥ 70 years was analysed. Newly diagnosed (n = 34) and refractory multiple myeloma (n = 36) patients were studied. The median age was 72 years (range: 70–82·6). CD34+ cells were mobilized with chemotherapy and granulocyte colony‐stimulating factor (G‐CSF) (n = 35) or G‐CSF alone (n = 35), yielding medians of 11·8 × 106 versus 8 × 106cells/kg respectively (P = 0·007). Because of excessive mortality (16%) in the first 25 patients who received melphalan 200 mg/m2 (MEL‐200), the dose was subsequently decreased to 140 mg/m2 (MEL‐140). Median times to absolute neutrophil count (ANC) > 0·5 × 109/l and to platelets > 20 × 109/l were 11 and 13 d respectively. Thirty‐one patients (44%) received tandem auto‐SCT. Complete remission (CR) was 20% after the first SCT and 27% after tandem SCT. Median CR duration was 1·5 years and was significantly longer for patients with ≤ 12 months of prior chemotherapy (2·6 versus 1·0 years, P = 0·0008). The 3‐year event‐free survival (EFS) and overall survival (OS) (+ standard error, SE) were projected at 20% + 9% and 31% + 10% respectively. Tandem SCTs positively affected EFS (4·0 versus 0·7 years; P = 0·003) and OS (4·0 versus 1·4 years; P = 0·02) compared with single auto‐SCT. In conclusion, MEL‐140 is less toxic and appears equally as efficacious as MEL‐200 in elderly patients. The benefits of tandem SCT in this patient population need further evaluation in a randomized trial.