bluuebunny/crossref_metadata_2025 huggingface dataset:
Complement activation plays a key role in the side‐effects of rituximab treatment
Authors: Van Der Kolk L. E. , Grillo‐López A. J. , Baars J. W. , Hack C. E. , Van Oers M. H. J.
Published: March 2003

Treatment with rituximab, a chimaeric anti‐CD20 monoclonal antibody, can be associated with moderate to severe first‐dose side‐effects, notably in patients with high numbers of circulating tumour cells. The aim of this study was to elucidate the mechanism of these side‐effects. At multiple early time points during the first infusion of rituximab, complement activation products (C3b/c and C4b/c) and cytokines [tumour necrosis factor alpha (TNF‐α), interleukin 6 (IL‐6) and IL‐8] were measured in five relapsed low‐grade non‐Hodgkin's lymphoma (NHL) patients. Infusion of rituximab induced rapid complement activation, preceding the release of TNF‐α, IL‐6 and IL‐8. Although the study group was small, the level of complement activation appeared to be correlated both with the number of circulating B cells prior to the infusion (r = 0·85; P = 0·07) and with the severity of the side‐effects. We conclude that complement plays a pivotal role in the pathogenesis of side‐effects of rituximab treatment. As complement activation can not be prevented by corticosteroids, it might be relevant to study the possible role of complement inhibitors during the first administration of rituximab.
Matrix metalloproteinase‐9 and vascular endothelial growth factor: a possible link in adult T‐cell leukaemia cell invasion
Authors: Hayashibara Toshihisa , Yamada Yasuaki , Onimaru Yasuyuki , Tsutsumi Chizuko , Nakayama Susumu , Mori Naoki , Miyanishi Takayuki , Kamihira Shimeru , Tomonaga Masao , Maita Tetsuo
Published: March 2003

Summary. Plasma from a total of 57 patients with adult T‐cell leukaemia (ATL) (acute ATL, 39 patients; lymphoma ATL, one patient; chronic ATL, 15 patients; smouldering ATL, two patients) and 20 healthy controls was analysed for the presence of type IV gelatinase activity with clinical features. A significant elevation of plasma matrix metalloproteinase‐9 (MMP‐9) was observed in some ATL patients, particularly in the patients with malignant cell infiltration. MMP‐9 was found to be secreted into the conditioned medium from all ATL cell lines examined. Moreover, the corresponding mRNA was detectable both in all ATL cell lines examined and in the majority of primary acute ATL cells, indicating that ATL cells are capable of synthesizing and secreting MMP‐9. We previously demonstrated that a high incidence of ATL cell infiltration was closely related to a high plasma level of vascular endothelial growth factor (VEGF) produced by ATL cells themselves. This present study showed that the presence of increased plasma MMP‐9 was closely associated with elevated plasma VEGF in ATL patients. Furthermore, we showed that both increased plasma MMP‐9 and VEGF were significantly related to high ATL cell infiltration. All these findings strongly suggest that MMP‐9 and VEGF act co‐operatively in the process of ATL cell invasion.
Perforin defects of primary haemophagocytic lymphohistiocytosis in Japan
Authors: Suga Naohiro , Takada Hidetoshi , Nomura Akihiko , Ohga Shouichi , Ishii Eiichi , Ihara Kenji , Ohshima Koichi , Hara Toshiro
Published: March 2003

Summary. The perforin gene was analysed in 15 Japanese patients with primary haemophagocytic lymphohistiocytosis (HLH). Perforin gene defects were found in two out of eight patients with familial HLH (FHL), and one out of seven without affected siblings. Four novel mutations were identified. Compound heterozygous mutations (one FHL and one sporadic HLH) and only one allele mutation (one FHL) were defined. Flow cytometry revealed no perforin expression in CD8+ or CD56+ cells from a surviving patient with a mutation. The frequency of mutation was at least 20% of FHL in Japan. Flow cytometry for intracellular perforin may be useful for the screening of FHL2.
Age, sex and vitamin status affect plasma level of homocysteine, but hyperhomocysteinaemia is possibly not an important risk factor for venous thrombophilia in Taiwanese Chinese
Authors: Lin Jen‐Shiou , Shen Ming‐Ching , Cheng Wern‐Cherng , Tsay Woei , Wang Ying‐Chi , Lin Bin‐Bin , Hung Mei‐Hua
Published: March 2003

Summary. The biological effects of age, sex and vitamin status on plasma total homocysteine (tHcy), and association of hyperhomocysteinaemia with venous thromboembolism in Taiwanese Chinese individuals, were investigated. Eighty patients (16–85 years) with venous thrombophilia and 123 healthy subjects (15–85 years) without history of vascular thrombosis were studied for plasma levels of tHcy, folate and vitamin B12. A multivariate analysis in healthy subjects revealed that plasma tHcy levels tended to increase with age (P < 0·001) and with decreasing plasma levels of folate (P=0·001) or vitamin B12 (P < 0·029); men tended to have higher plasma tHcy levels than women (P=0·006). Thrombotic risk assessment in a case–control study demonstrated that neither plasma level of tHcy [odds ratio (OR), 1·07; 95% confidence interval (CI), 0·96–1·18; P=0·210] nor hyperhomocysteinaemia (OR, 1·65; 95% CI, 0·50–5·49; P=0·415) was significantly associated with venous thrombophilia. The relationship between hyperhomocysteinaemia and recurrence of episode remained insignificant (P=0·560). We conclude that age, sex and vitamin status affect plasma tHcy but hyperhomocysteinaemia is possibly not an important risk factor for venous thrombophilia in Taiwanese Chinese.
Abdominal infections in patients with acute leukaemia: a prospective study applying ultrasonography and microbiology
Authors: Gorschlüter Marcus , Marklein Günter , Höfling Katja , Clarenbach Ricarda , Baumgartner Stefanie , Hahn Corinna , Ziske Carsten , Mey Ulrich , Heller Ricarda , Eis‐Hübinger Anna Maria , Sauerbruch Tilman , Schmidt‐Wolf Ingo G. H. , Glasmacher Axel
Published: March 2003

Summary.  A prospective study of 62 chemotherapy‐ induced neutropenic episodes in patients with acute leukaemia was conducted to determine the incidence and causes of abdominal infections, and to assess the diagnostic value of the combined use of ultrasonography (US) and microbiology. Each patient underwent US of liver, gallbladder and complete bowel before chemotherapy, on days 2–4 after the end of chemotherapy and in cases of fever, diarrhoea or abdominal pain. US was combined with a standardized clinical examination and a broad spectrum of microbiological investigations. From January to August 2001, 243 US examinations were performed. The overall incidence of abdominal infectious diseases was 17·7% (11 out of 62, 95% confidence interval (CI): 9–29%). Four patients (6·5%) developed neutropenic enterocolitis; two of them died, two survived. Bowel wall thickening (BWT) > 4 mm in these four patients ranged from 5·8 to 23·6 mm and was detected only in one patient with mucositis. In three other patients (4·8%) Clostridium difficile, and in one patient (1·6%) Campylobacter jejuni, caused enterocolitis without BWT. Cholecystitis was diagnosed in three patients (4·8%) and hepatic candidiasis was strongly suspected in one patient. Abdominal infections caused by gastroenteritis viruses, cytomegalovirus (CMV) or Cryptosporidium were not observed. We conclude that in neutropenic patients with acute leukaemia receiving chemotherapy: (i) BWT is not a feature of chemotherapy‐induced mucositis and should therefore be considered as sign of infectious enterocolitis; (ii) viruses, classic bacterial enteric pathogens (Salmonella, Shigella, Yersinia, Campylobacter, Aeromonas, Vibrio subsp., enterohaemorrhagic Escherichia coli) and Cryptosporidium have a very low incidence; and (iii) abdominal infections may be underestimated when US is not used in every patient with abdominal pain.
Red blood cells from patients with homozygous sickle cell disease provide a catalytic surface for factor Va inactivation by activated protein C
Authors: Bezeaud Annie , Venisse Laurence , Helley Dominique , Trichet Catherine , Girot Robert , Guillin Marie‐Claude
Published: March 2003

Summary.  The structure of red blood cell (RBC) membranes in homozygous sickle cell disease (SCD) is significantly disturbed, with an increased exposure of aminophospholipids (phosphatidylserine and phosphatidylethanolamine) at the outer surface, responsible for a procoagulant activity of SS RBCs. Aminophospholipids are known not only to promote procoagulant reactions, but also to support inhibition of blood coagulation by the protein C system. The aim of the present study was to examine whether SS RBCs could serve as a catalytic surface for the inactivation of factor Va by activated protein C (APC). Venous blood was obtained from 19 consecutive SS patients and 13 controls (AA). In all SS patients, the amount of phosphatidylserine exposed at the outer surface of RBCs was increased compared with controls, as demonstrated by a prothrombinase assay. In addition, SS RBCs significantly (P < 0·0001) increased the rate of FVa inactivation by APC: the mean values (and ranges) of the factor Va inactivation rates were 30 (0–57) vs 9·5 (0–32) mmol Vai/min/mol APC for SS RBCs and normal RBCs respectively. Our results indicate that SS RBCs provide a catalytic surface for the negative control of blood coagulation, which may partially control the procoagulant activity of these cells.
Anaplastic large cell lymphoma in childhood: analysis of 72 patients treated on The United Kingdom Children's Cancer Study Group chemotherapy regimens
Authors: Williams Denise M. , Hobson Rachel , Imeson John , Gerrard Mary , McCarthy Keith , Pinkerton C. Ross ,   The United Kingdom Children's Cancer Study Group
Published: March 2003

Summary.  From June 1990 to June 1998, 72 patients with anaplastic large cell lymphoma (ALCL) were treated with short intensive multi‐agent regimens [non‐Hodgkin's lymphoma (NHL) 9000 and 9602]. Diagnosis was based on morphological and immunophenotypic criteria. Treatment for stage I disease consisted of eight courses (2 × vincristine, doxorubicin, prednisolone; 2 × methotrexate; 2 × cytarabine, thioguanine; and 2 × methotrexate etoposide). For stage II, III and non‐central nervous system (CNS) stage IV, two COPADM (cyclophosphamide, doxorubicin, prednisolone, methotrexate, vincristine), two CYM (cytarabine methotrexate) and a COPADM was given. For CNS‐positive disease, treatment was intensified and contained methotrexate 8 g/m2 and cytarabine 3 g/m2. Fifty‐nine patients (82%) achieved a remission. Thirteen of these relapsed, with a median time to relapse from the start of treatment of 5 months (range 3–14). Relapse included a new site in 9/13 patients. The probabilities of overall and event free survival at 5 years were 65% (53–76%) and 59% (47–70%), respectively, with a median follow up of 4·3 years. Mediastinal and visceral involvement at presentation were found to be predictive of an increased risk of failure.
Functional consequences of perforin gene mutations in 22 patients with familial haemophagocytic lymphohistiocytosis
Authors: Feldmann Jérôme , Le Deist Françoise , Ouachée‐Chardin Marie , Certain Stéphanie , Alexander Sarah , Quartier Pierre , Haddad Elie , Wulffraat Nico , Casanova Jean Laurent , Blanche Stéphane , Fischer Alain , De Saint Basile Geneviève
Published: March 2003

Summary. Familial haemophagocytic lymphohistiocytosis (FHL), an inherited form of haemophagocytic lymphohistiocytosis (HLH) syndrome, is characterized by the overwhelming activation of T lymphocytes and macrophages invariably leading to death in the absence of treatment. FHL is a heterogeneous autosomal recessive disorder, with one known causative gene which codes for perforin, a cytotoxic effector protein. In this study, we have characterized the genotype and phenotype of 14 unrelated families with perforin deficiency. Four new missense mutations of the perforin gene were identified. In every case, perforin gene mutations led to undetectable intracellular perforin expression within cytotoxic cells, while some residual T‐cell cytotoxic activity could be associated with certain missense mutations. Clinical and biological analyses did not differentiate between patients with nonsense or missense mutations, although age at diagnosis, which tended to be similar within members of the same family, was delayed in patients from two families belonging to the second group. In one case, consequences of perforin deficiency, diagnosed at birth, could be assessed prior to onset of clinical manifestations. No evidence for T‐cell activation could be shown, suggesting that an exogenous event is required to trigger the disease manifestation. Control assessment of perforin expression and cytotoxic assays by lymphocytes from young children led to the conclusion that perforin content of natural killer cells could be a reliable diagnostic test at any age. Altogether, these data enabled a better characterization of perforin deficiency and its consequences, and defined reliable diagnostic tools.
Association between xenobiotic gene polymorphisms and non‐Hodgkin's lymphoma risk
Authors: Kerridge Ian , Lincz Lisa , Scorgie Fiona , Hickey Danica , Granter Neil , Spencer Andrew
Published: March 2003

Summary. The last four decades have seen a significant increase in the incidence of non‐Hodgkin's lymphoma (NHL) as a possible result of increasing environmental carcinogen exposure, particularly pesticides and solvents. Based on the increasing evidence for an association between carcinogen exposure‐related cancer risk and xenobiotic gene polymorphisms, we have undertaken a case–control study of xenobiotic gene polymorphisms in individuals with a diagnosis of NHL. Polymorphisms of six xenobiotic genes (CYP1A1, GSTT1, GSTM1, PON1, NAT1, NAT2) were characterized in 169 individuals with NHL and 205 normal controls using polymerase chain reaction‐based methods. Polymorphic frequencies were compared using Fisher's exact tests, and odds ratios for NHL risk were calculated. Among the NHL group, the incidence of GSTT1 null and PON1 BB genotypes were significantly increased compared with controls, 34%vs 14%, and 24%vs 11% respectively. Adjusted odds ratios calculated from multivariate analyses demonstrated that GSTT1 null conferred a fourfold increase in NHL risk (OR = 4·27; 95% CI, 2·40–7·61, P < 0·001) and PON1 BB a 2·9‐fold increase (OR = 2·92; 95% CI, 1·49–5·72, P = 0·002). Furthermore, GSTT1 null combined with PON1 BB or GSTM1 null conferred an additional risk of NHL. This is the first time that a PON1 gene polymorphism has been shown to be associated with cancer risk. We conclude that the two polymorphisms, GSTT1 null and PON1 BB, are common genetic traits that pose low individual risk but may be important determinants of overall population NHL risk, particularly among groups exposed to NHL‐related carcinogens.
The membrane characteristics of Plasmodium falciparum‐infected and ‐uninfected heterozygous α0thalassaemic erythrocytes
Authors: Williams Thomas N. , Weatherall David J. , Newbold Christopher I.
Published: March 2003

Summary. The α thalassaemias are the commonest known human genetic disorders. Although they have almost certainly risen to their current frequencies through natural selection by malaria, the precise mechanism of malaria protection remains unknown. We have investigated the characteristics of red blood cells (RBCs) from individuals heterozygous for α0thalassaemia (–/αα) from a range of perspectives. On the basis of the hypothesis that defects in membrane transport could be relevant to the mechanism of malaria protection, we investigated sodium and potassium transport and the activity of the Plamodium falciparum‐induced choline channel but found no significant differences in –/αα RBCs. Using flow cytometry, we found that thalassaemic P. falciparum‐infected RBCs (IRBCs) bound 44% more antibody from immune plasma than control IRBCs. This excess binding was abrogated by predigestion of IRBCs with trypsin but was not directed at the variant surface molecule PfEMP1. Furthermore, we found no evidence for altered cytoadhesion of α‐thalassaemic IRBCs to the endothelial receptors intercellular adhesion molecule‐1 (ICAM‐1), CD36 or thrombospondin. We hypothesize that altered red‐cell membrane band 3 protein may be a target for enhanced antibody binding to α‐thalassaemic IRBCs and could be involved in the mechanism of malaria protection.
A Tyr346→Cys substitution in the interdomain acidic region a1 of factor VIII in an individual with factor VIII:C assay discrepancy
Authors: Mumford Andrew D. , Laffan Michael , O'Donnell James , McVey John H. , Johnson Daniel J. D. , Manning Richard A. , Kemball‐Cook Geoffrey
Published: March 2003

Summary. The interdomain acidic region a1 is a unique structural feature of coagulation factor VIII (FVIII) and may mediate the proteolytic activation of FVIII and the inactivation of FVIIIa. We report an individual with a Tyr346→Cys substitution within region a1, who presented with a one‐stage FVIII activity (FVIII:C) of 0·34 iu/ml (normal range 0·5–2·0) but normal two‐stage FVIII:C and FVIII antigen values. In a factor Xa (FXa)‐generation assay for FVIII in which the activation time with thrombin was varied, the variant plasma showed normal FVIII:C at both short and long activation times. However, at intermediate activation times the FXa generation of the variant plasma was less than that of normal pooled plasma. In a modified one‐stage FVIII:C assay in which partially purified FVIII was activated with thrombin at low concentrations, the variant FVIII showed less activation than wild‐type FVIII, although this defect corrected with increasing concentrations of thrombin. When partially purified variant FVIII was activated with a large molar excess of thrombin, the subsequent rate of decay of FVIII:C was greater for variant FVIII. The complex defects in activation and inactivation displayed by FVIII Tyr346→Cys support the hypothesis that the a1 sequence is a key regulator of FVIII activity.
A molecular study of the t(4;14) in multiple myeloma
Authors: Sibley Kathryn , Fenton James A. L. , Dring Ann M. , Ashcroft Andrew J. , Rawstron Andrew C. , Morgan Gareth J.
Published: March 2003

Summary. The t(4;14) translocation is found in approximately 10% of myeloma patients and results in the deregulation of at least two genes, MMSET and fibroblast growth factor receptor 3 (FGFR3), with the formation of a fusion product between MMSET and the immunoglobulin heavy chain (IgH) locus and overexpression of FGFR3. We have analysed a series of 80 patient samples, comprising 67 multiple myeloma (MM) cases and 13 monoclonalgammopathy of undetermined significance (MGUS) cases, using RT–PCR to detect IgH–MMSET fusions. The t(4;14) translocation was detected in 7/67 (10%) myeloma cases and all seven expressed FGFR3 which was not seen in t(4;14)‐negative myeloma cases. In the MGUS cases, a similar proportion of t(4;14)‐positive cases was found (2/13; 15%), but none of these expressed FGFR3. All patients with detectable FGFR3 expressed both the FGFR3 IIIb and FGFR3 IIIc isoforms, the result of alternative splicing in the ligand binding domain, and exon‐deleted variants of FGFR3. We also identified a cryptic splice site in MMSET which results in a 277 amino acid deletion downstream of the breakpoint on der(4). FGFR3 mutation analysis revealed no mutations in the presenting myeloma or MGUS samples. However, we also had access to paired presentation and relapse samples which had been taken from a patient 13 months apart. Both samples had the t(4;14) translocation and overexpressed FGFR3, but only the relapse sample possessed the K650E mutation in the kinase domain of FGFR3. This suggests that targeted mutation in the translocated FGFR3 gene when under the control of the immunoglobulin promoters can occur and may provide one mechanism for disease progression.
Microvessel density, a surrogate marker of angiogenesis, is significantly related to survival in multiple myeloma patients
Authors: Pruneri Giancarlo , Ponzoni Maurilio , Ferreri Andrés J. M. , Decarli Nicola , Tresoldi Moreno , Raggi Francesca , Baldessari Chiara , Freschi Massimo , Baldini Luca , Goldaniga Maria , Neri Antonino , Carboni Nadia , Bertolini Francesco , Viale Giuseppe
Published: March 2003

Summary. We evaluated microvessel density (MVD) in bone marrow biopsies (BM) from multiple myeloma (MM) patients after staining with anti‐CD34 and anti‐CD105 antibodies (mAbs). The anti‐CD105 mAb was significantly more sensitive than the anti‐CD34 mAb in visualizing blood vessels both in controls and MM samples. MVD was significantly higher in MM than in controls with both anti‐CD34 and anti‐CD105 mAbs. Patients with low CD34+ MVD survived longer than patients with higher MVD (P = 0·01), whereas there was no difference in survival between patients with low and high CD105+ MVD. Multivariate analysis confirmed the independent significant association between CD34+ MVD and survival (P = 0·001).
Mutation spectrum in Australian pedigrees with hereditary hyperferritinaemia–cataract syndrome reveals novel and de novo mutations
Authors: McLeod Janet L. , Craig Jamie , Gumley Sarah , Roberts Sarah , Kirkland Mark A.
Published: March 2003

Summary.  Hereditary hyperferritinaemia–cataract syndrome (HHCS) (OMIM #600886) is a rare autosomal dominant condition identified by high serum ferritin levels with normal iron saturation and distinctive bilateral cataract. It may be misdiagnosed as haemochromatosis and such patients become anaemic as a result of inappropriate venesection. The elevated serum ferritin is due to a mutation in the iron‐responsive element (IRE) of the l‐ferritin gene, resulting in excessive l‐ferritin production. We report the identification of three Australian pedigrees; one with a previously described mutation at position 40, a pedigree with a novel mutation at position 39 and an individual with a de novo mutation at position 32 of the l‐ferritin IRE.
A single high dose of idarubicin combined with high‐dose ARA‐C for treatment of first relapse in childhood ‘high‐risk’ acute lymphoblastic leukaemia: a study of the AIEOP group
Authors: Testi Anna Maria , Del Giudice Ilaria , Arcese William , Moleti Maria Luisa , Giona Fiorina , Basso Giuseppe , Biondi Andrea , Conter Valentino , Messina Chiara , Rondelli Roberto , Micozzi Alessandra , Micalizzi Concetta , Barisone Elena , Locatelli Franco , Dini Giorgio , Aricò Maurizio , Casale Fiorina , Comis Margherita , Ladogana Saverio , Lippi Alma , Mura Rossella , Pinta Marie France , Santoro Nicola , Valsecchi Maria Grazia , Masera Giuseppe , Mandelli Franco
Published: March 2003

Summary. The outcome of children with acute lymphoblastic leukaemia (ALL) and early relapse remains unsatisfactory. In January 1995, the AIEOP (Associazione Italiana di Oncologia ed Ematologia Pediatrica) group opened a trial for children with ALL in first isolated or combined bone marrow relapse defined at high risk according to the length of first remission and the immunophenotype. The treatment plan included the combination of a single high‐dose idarubicin and high‐dose cytarabine as induction therapy followed by an intensive consolidation and stem cell transplant (SCT). In total, 100 children from 16 Italian centres were enrolled; 80 out of the 99 evaluable patients (81%) achieved second complete remission; eight (8%) died during induction and 11 (11%) failed to respond. A total of 42 out of the 80 responders (52·5%) received a SCT: 19 from an identical sibling, 11 from a matched unrelated donor and 12 from umbilical cord blood cells. The estimated 4‐year overall survival and event‐free survival were 25% and 21% respectively. Disease‐free survival at 4 years was 25·8% for the 80 responders. At 4 years, 39 out of 100 children remain alive, with 27 of them free of leukaemia. This induction therapy has shown antileukaemic efficacy with acceptable toxicity; moreover, all responders proved eligible for intensive consolidation.
High P‐glycoprotein‐mediated export observed in patients with a history of idiopathic thrombocytopenic purpura
Authors: Levy Adam S. , Cunningham‐Rundles Susanna , Mazza BethAnne , Simm Maciej , Gorlick Richard , Bussel James
Published: March 2003

Summary.  Studies have suggested that high P‐glycoprotein expression in lymphocytes from patients with autoimmune disorders may affect disease outcome. Idiopathic thrombocytopenic purpura (ITP) and Evans' syndrome are widely thought to be autoimmune processes, however, the precise mechanisms remain unknown. Peripheral blood mononuclear cells from patients with refractory or recurrent ITP or Evans' syndrome were studied using the rhodamine 123 flow cytometric assay to investigate functional export levels. Lymphocytes from ITP and Evans' syndrome patients showed a significantly decreased ability to retain rhodamine, suggesting increased export protein function. Reverse transcription polymerase chain reaction distinguished P‐glycoprotein as the likely export protein.
Thrombotic thrombocytopenic purpura after allogeneic stem cell transplantation: a survey of the European Group for Blood and Marrow Transplantation (EBMT)
Authors: Ruutu Tapani , Hermans Jo , Niederwieser Dietger , Gratwohl Alois , Kiehl Michael , Volin Liisa , Bertz Harmut , Ljungman Per , Spence David , Verdonck Leo F. , Prentice H. Grant , Bosi Alberto , Du Toit Cecile E. , Brinch Lorentz , Apperley Jane F. ,   the EBMT Chronic Leukaemia Working Party
Published: March 2003

Summary. A survey was carried out among the European Group for Blood and Marrow Transplantation (EBMT) centres to determine the incidence, risk factors, treatment and outcome of thrombotic thrombocytopenic purpura (TTP) following allogeneic haematopoietic stem cell transplantation. TTP was defined as the simultaneous occurrence of red cell fragmentation, laboratory findings of haemolysis, red cell transfusion requirement and de novo or persistant thrombocytopenia caused by consumption, in the absence of disseminated intravascular coagulation. Forty‐five centres reported all patients (n = 406) transplanted between July and December 1996. Twenty‐three patients developed TTP; the risk of developing TTP was 6·7% at 2 years (95% CI: 4·1% to 9·3%). The median time of onset was 44 d (range 13–319) post transplantation. Significant risk factors for the development of TTP were female gender (P = 0·005) and an unrelated donor (P = 0·046). To treat TTP, cyclosporin administration was discontinued in 10 cases, plasma exchanges were performed in five cases and 12 patients received plasma infusions without plasma exchange. TTP resolved in 13 of the 23 patients (57%). The only factor predictive of resolution of TTP was the absence of nephropathy. Seven patients (30%) were alive at follow‐up of 38–45 months from the onset of TTP. Sixteen patients died; the causes were multiple, only three patients had TTP as a central factor. The median time to death was 41 d (range 1–762 d) from the onset of TTP. TTP is a relatively frequent complication of allogeneic stem cell transplantation and it is associated with high mortality, though death is usually caused by multiple factors.
Enhanced killing of B lymphoma cells by granulocyte colony‐stimulating factor‐primed effector cells and Hu1D10 – a humanized human leucocyte antigen DR antibody
Authors: Stockmeyer Bernhard , Schiller Martin , Repp Roland , Lorenz Hanns‐Martin , Kalden Joachim R. , Gramatzki Martin , Valerius Thomas
Published: March 2003

Summary. Antibody‐based approaches have become a novel treatment modality for lymphoma patients. Humanized 1D10 (Hu1D10; Remitogen) is among the antibodies that are currently under evaluation in phase II clinical trials in lymphoma patients. The 1D10 antibody is directed against a polymorphic epitope on the β‐chain of human leucocyte antigen (HLA) class II. We found expression of the 1D10 epitope on B cells and monocytes from approximately 50% of healthy donors. Analyses of 1D10 expression on malignant cells revealed that approximately half of the HLA class II‐positive haematological malignancies expressed the 1D10 epitope. In whole blood antibody‐dependent cellular cytotoxicity (ADCC) assays, Hu1D10 was more effective than rituxan in killing malignant ARH‐77 B cells. Interestingly, Hu1D10‐mediated lymphoma cell lysis was significantly enhanced when blood from granulocyte colony‐stimulating factor (G‐CSF)‐treated patients was compared with blood from healthy controls. Analyses of the relevant effector cell populations revealed that FcγRI (CD64)‐positive polymorphonuclear cells were critical for enhanced Hu1D10‐mediated lymphoma killing during G‐CSF therapy, while the same effector cell population induced only marginal lysis with rituxan. Furthermore, Hu1D10 was highly effective in inducing apoptosis in primary lymphoma cells from B chronic lymphocytic leukaemia patients. These preclinical results form the basis for a phase I/II clinical trial of Hu1D10 in combination with G‐CSF.
Detection of lupus anticoagulant identifies patients with autoimmune haemolytic anaemia at increased risk for venous thromboembolism
Authors: Pullarkat Vinod , Ngo Mark , Iqbal Syma , Espina Byron , Liebman Howard A.
Published: March 2003

Summary. Venous thromboembolism (VTE) is a well‐recognized complication of autoimmune haemolytic anaemia (AIHA), and is a major cause of morbidity and mortality in this disorder. However, the incidence, pathogenesis and risk‐factors for VTE in AIHA remain poorly defined. Lupus anticoagulants (LA) and anticardiolipin (ACA) antibodies are autoantibodies directed against epitopes on prothrombin or β2 glycoprotein I (β2‐GPI). Both LA and ACA (together called antiphospholipid antibodies, APLA) are associated with VTE. We have prospectively studied the occurrence of VTE and APLA in 30 patients with AIHA. VTE was objectively documented in eight (27%) patients. APLA were detected in 19 (63%) patients with AIHA, of whom nine (30%) had a LA and 17 (57%) ACA. Seven patients had both LA and ACA. Among the eight patients with VTE, LA was detected in five (63%) and ACA in four (50%). There was a statistically significant association between presence of LA and occurrence of VTE (RR: 7·50, 95% CI: 1·25–45·2, P = 0·03). VTE is a frequent and life‐threatening complication of AIHA. Detection of the lupus anticoagulant in patients with AIHA identifies individuals at significantly increased risk for VTE. Future studies should address the role of prophylactic anticoagulation in patients with AIHA.
Isolated parenchymal lung involvement in children with stage IV Hodgkin's disease: results of the UKCCSG HD8201 and HD9201 studies
Authors: Atra Ayad , Higgs Elizabeth , Capra Michael , Elsworth Ann , Imeson John , Radford Martin , Pinkerton Ross , Hewitt M. ,   on behalf of the United Kingdom Children Cancer Study Group (UKCCSG)/Hodgkin's Disease Group
Published: March 2003

Summary. We retrospectively reviewed the case notes of 27 patients who were diagnosed with stage IV Hodgkin's disease (HD) because of isolated parenchymal pulmonary involvement on chest radiograph and computerized tomography scan (excluding subcategory E). Ten were boys and 15 had B symptoms. Median age at diagnosis was 13·6 years (range 6·1–16·2). All received 6–8 cycles of ChlVPP (chlorambucil, vinblastine, procarbazine and prednisolone) and two had additional whole lung irradiation (12 Gy). Ten patients (37%) relapsed or progressed. Seven survive following second‐line therapy while three died, two of HD and one of secondary acute myeloid leukaemia 4 years from diagnosis. At the time of analysis, the median follow‐up of patients was 56 months (range 9–127). The event‐free survival (EFS) was 58·4% (95% CI 38·5–75·8%) at both 5 and 10 years from diagnosis, and the overall survival (OS) was 84·2% (95% CI 61·8–94·6%) at both 5 and 10 years from diagnosis. We conclude that the outcome for HD patients defined as stage IV, because of isolated parenchymal lung involvement, is encouraging and compares favourably with other extra lymphatic organ involvement. Combination chemotherapy is effective in achieving long‐term remission and whole lung irradiation is unnecessary.
Specificity of polymerase chain reaction‐based clonality analysis of immunoglobulin heavy chain gene rearrangement for the detection of bone marrow infiltrate in B‐cell lymphoma‐associated haemophagocytic syndrome
Authors: Kojima Kensuke , Kaneda Kinuyo , Yasukawa Masaki , Tanaka Kazuo , Inoue Takeshi , Yamashita Takuya , Dansako Hiromichi , Sakugawa Sumie Takase , Kozuka Teruhiko , Hara Masamichi , Tanimoto Mitsune
Published: March 2003

Summary. As a wide range of disorders underlie haemophagocytic syndrome, a rapid distinction between benign polyclonal and malignant monoclonal lymphoid proliferations is critical. We investigated whether polymerase chain reaction (PCR) amplification of immunoglobulin heavy chain gene rearrangement could efficiently detect clonal B‐cell populations in non‐diagnostic marrow for B‐cell lymphoma‐associated haemophagocytic syndrome (B‐LAHS). On amplifying two DNA samples per biopsy, no reproducible monoclonal PCR result was found in reactive haemophagocytic marrows. In contrast, four out of nine assessable B‐LAHS patients with histomorphologically and immunohistochemically lymphoma‐free bone marrow showed a reproducible monoclonal immunoglobulin heavy chain gene rearrangement. At the molecular level, two B‐LAHS patients had lymphoma‐free marrow as demonstrated by patient‐specific PCR, suggesting that haemophagocytic marrow is not always associated with lymphoma involvement. PCR‐based demonstration of clonal B‐cell populations in marrow would add an extra dimension to B‐LAHS diagnosis.
C‐reactive protein in the diagnosis of deep vein thrombosis
Authors: Bucek Robert A. , Reiter Markus , Quehenberger Peter , Minar Erich
Published: March 2003

Summary. The role of C‐reactive protein (CRP) in the diagnosis of suspected deep vein thrombosis (DVT) and a possible advantage of its additional evaluation with D‐dimer has not been clearly evaluated. We therefore studied plasma CRP and D‐dimer levels in 233 consecutive patients with suspected DVT; the final diagnosis was based on the results of colour duplex ultrasound or venography. DVT was diagnosed in 31·3%. CRP and D‐dimer correlated significantly (r = 0·64, P < 0·01); both were increased significantly in patients suffering from DVT (P < 0·001). Multivariate analysis revealed a significant influence of the presence of DVT (P < 0·001), the presence of malignancy (P < 0·001) and the presence of inflammatory diseases (P = 0·009) on plasma CRP, while there was no significant influence of the duration of symptoms (P = 0·30). The sensitivity (75%vs 93%) to specificity (69%vs 55%) relationship showed inferior results for CRP compared with D‐dimer; its additional evaluation did not improve the diagnostic value of D‐dimer. We conclude that CRP can provide additional information neither for the diagnostic process in patients with suspected DVT nor for the differential diagnosis of DVT and inflammatory diseases.
Hepatocyte growth factor in myeloma patients treated with high‐dose chemotherapy
Authors: Seidel Carina , Lenhoff Stig , Brabrand Sigmund , Anderson Gøran , Standal Therese , Lanng‐Nielsen Johan , Turesson Ingemar , Børset Magne , Waage Anders ,   for the Nordic Myeloma Study Group
Published: March 2003

Summary. Hepatocyte growth factor (HGF) is a cytokine produced by myeloma cells. We examined serum HGF levels in a population of young myeloma patients (median age 52 years) treated with high‐dose chemotherapy. Sera from 128 myeloma patients at diagnosis and serial samples from 16 patients were analysed. Compared with 62 healthy controls, HGF was elevated at diagnosis in 25% of patients (median 0·48 and 1·08 ng/ml respectively; P < 0·0001). The 95 patients who completed therapy were analysed for the impact of HGF on survival. Median survival was not reached after 77 months in the patient group with normal HGF values (< 1·7 ng/ml, n = 69). In the group with elevated HGF (≥ 1·7 ng/ml, n = 26), median survival was 63 months (P = 0·08). In 16 patients, serum was drawn at diagnosis and at the time of expected disease remission (6 weeks to 3 months after chemotherapy). HGF values declined after treatment in 14 of these patients, from a median of 0·9 ng/ml (0·49–1·65) to 0·42 ng/ml (0·32–0·73) (P = 0·005). Our results show that in young myeloma patients HGF is elevated, and that patients with higher levels had a trend towards poorer prognosis. Treatment with high‐dose chemotherapy reduced HGF in the serum of the majority of patients.
Investigatory and analytical approaches to differential gene expression profiling in mantle cell lymphoma
Authors: Zhu Ying , Hollmén Jaakko , Räty Riikka , Aalto Yan , Nagy Balint , Elonen Erkki , Kere Juha , Mannila Heikki , Franssila Kaarle , Knuutila Sakari
Published: March 2003

Summary. Mantle cell lymphoma (MCL) is a non‐Hodgkin's lymphoma of B‐cell lineage. The blastoid variant of MCL, characterized by high mitotic rate, is clinically more aggressive than common MCL. We used the cDNA array technology to examine the gene expression profiles of both blastoid variant and common MCL. The data was analysed by regression analysis, principal component analysis and the naive Bayes' classifier. Eight genes were identified as differentially deregulated between the two groups. Oncogenes CMYC, BCL2 and PIM1 were upregulated more frequently in the blastoid variant than in common MCL. This implied that the gp130‐mediated signal transducer and activator of transcription 3 (STAT3) signalling pathway was involved in the blastoid variant transformation of MCL. Other differentially deregulated genes were TOP1, CD23, CD45, CD70 and NFATC. By using the eight differentially deregulated genes, we created a classifier to distinguish the blastoid variant from common MCL with high accuracy. We also identified 18 genes that were deregulated in both groups. Among them, BCL1, CALLA/CD10 and GRN were suggested to be oncogenes. The products of RGS1, RGS2, ANX2 and CD44H were suggested to promote tumour metastasis. CD66D was suggested to be a tumour suppressor gene.
Results and follow‐up of a phase III randomized study of recombinant human‐granulocyte stimulating factor as support for immunosuppressive therapy in patients with severe aplastic anaemia
Authors: Gluckman Eliane , Rokicka‐Milewska Riitta , Hann Ian , Nikiforakis Emmanouel , Tavakoli Filipos , Cohen‐Scali Sophie , Bacigalupo Andrea ,   for the European Group for Blood and Marrow Transplantation Working Party for Severe Aplastic Anemia
Published: March 2003

Summary. In patients with idiopathic severe aplastic anaemia who are treated with immunosuppressive agents to combat T lymphocyte‐mediated destruction of haematopoietic progenitor cells, neutropenia is a major cause of infections and toxicity. Evidence from preliminary studies suggests that recombinant human glycosylated granulocyte colony‐stimulating factor (lenograstim) increases the number and functionality of neutrophils in patients with severe aplastic anaemia. This randomized, parallel‐group, multicentre study was conducted to evaluate the efficacy and safety of subcutaneous lenograstim during the first 12 weeks of standard immunosuppressive therapy in 102 patients with de novo severe aplastic anaemia. The addition of lenograstim to standard therapy resulted in an increase in the proportion of patients showing complete neutrophil response (83·0%vs 44·9%; P < 0·0001). This was seen even among patients with very severe aplastic anaemia (69·2%vs 31·6%; P = 0·012). In patients receiving lenograstim, median time to complete neutrophil response was shorter (6·3 vs 16·1 weeks; P = 0·0001) and mean duration of first neutrophil response was longer (P = 0·0248) than in the control group. At a median follow‐up of 5 years, no difference was observed between the groups in term of survival, haematological response and occurrence of secondary leukaemia (one patient in each group). We conclude that lenograstim support of immunosuppressive therapy might be used for patients with severe aplastic anaemia as it significantly enhances neutrophil recovery but does not modify the overall response and survival.
Fludarabine, cyclophosphamide and mitoxantrone in the treatment of resistant or relapsed chronic lymphocytic leukaemia
Authors: Bosch Francesc , Ferrer Anna , López‐Guillermo Armando , Giné Eva , Bellosillo Beatriz , Villamor Neus , Colomer Dolors , Cobo Francesc , Perales María , Esteve Jordi , Altés Albert , Besalduch Joan , Ribera Josep M. , Montserrat Emili ,   for the GELCAB (Grup per l'Estudi dels Limfomes a Catalunya i Balears),   Spain
Published: March 2003

Summary. We evaluated the efficacy and toxicity of fludarabine combined with cyclophosphamide and mitoxantrone (FCM) in patients with relapsed or resistant chronic lymphocytic leukaemia (CLL). In total, 37 patients with recurrent or resistant CLL received FCM: fludarabine 25 mg/m2 intravenously (IV), d 1–3; cyclophosphamide 200 mg/m2 IV, d 1–3; and mitoxantrone 6 mg/m2 IV, d 1, at 4‐week intervals for up to six courses. Moreover, 23 patients received FCM with cyclophosphamide 600 mg/m2 i.v. and mitoxantrone 8 mg/m2 i.v. on d 1. In addition to clinical methods, response was assessed using cytofluorometric and molecular techniques. ‘In vitro’ sensitivity to the FCM regimen was also analysed in 20 samples. The median number of courses given was 3 (range: 1–6). Overall, 30 patients (50%) achieved complete response (CR), including 10 cases of negative minimal residual disease (MRD(–)) (17%), and 17 (28%) partial response (PR). The median duration of response was 19 months. ‘In vitro’ sensitivity also correlated with CR achievement (P = 0·04). Main toxicity consisted of neutropenia, infections (8% of courses), and nausea and vomiting. The treatment‐related mortality was 5%. FCM did not hamper stem cell harvesting in patients who were candidates for autologous stem cell transplantation. FCM induced a high CR rate, including an important number of MRD(–), in patients with previously treated CLL.
Pharmacokinetics and red cell utilization of 52Fe/59Fe‐labelled iron polymaltose in anaemic patients using positron emission tomography
Authors: Beshara Soheir , Sörensen Jens , Lubberink Mark , Tolmachev Vladimir , Långström Bengt , Antoni Gunnar , Danielson Bo G. , Lundqvist Hans
Published: March 2003

Summary. Parenteral iron–polysaccharide complexes are increasingly applied. The pharmacokinetics of iron sucrose have been assessed by our group using positron emission tomography (PET). A single intravenous injection of 100 mg iron as iron (III) hydroxide–polymaltose complex, labelled with a tracer in the form of 52Fe/59Fe, was similarly assessed in six patients using PET for about 8 h. Red cell utilization was followed for 4 weeks. Iron polymaltose was similarly distributed to the liver, spleen and bone marrow. However, a larger proportion of this complex was rapidly distributed to the bone marrow. The shorter equilibration phase for the liver, about 25 min, indicates the minimal role of the liver for direct distribution. Splenic uptake also reflected the reticuloendothelial handling of this complex. Red cell utilization ranged from 61% to 99%. Despite the relatively higher uptake by the bone marrow, there was no saturation of marrow transport systems at this dose level. In conclusion, high red cell utilization of iron polymaltose occurred in anaemic patients. The major portion of the injected dose was rapidly distributed to the bone marrow. In addition, the reticuloendothelial uptake of this complex may reflect the safety of polysaccharide complexes. Non‐saturation of transport systems to the bone marrow indicated the presence of a large interstitial transport pool, which might possibly be transferrin.
Cytogenetic heterogeneity of acute myeloid leukaemia (AML) with trilineage dysplasia: Japan Adult Leukaemia Study Group‐AML 92 study
Authors: Miyazaki Yasushi , Kuriyama Kazutaka , Miyawaki Shuichi , Ohtake Shigeki , Sakamaki Hisashi , Matsuo Tatsuki , Emi Nobuhiko , Kobayashi Tohru , Matsushima Takafumi , Shinagawa Katsuji , Ohno Ryuzo , Tomonaga Masao ,   for the Japan Adult Leukaemia Study Group
Published: March 2003

Summary. Acute myeloid leukaemia (AML) with trilineage dysplasia (AML/TLD) is de novo AML recognized by the morphological dysplasia of three mature cell lines in the presence of leukaemic blasts. We studied the karyotypes of AML/TLD of patients with de novo AML, except for those with the French–American–British classification M3, who were enrolled onto the Japan Adult Leukaemia Study Group (JALSG)‐AML 92 trial. Morphological and cytogenetic analyses were performed in 559 patients and TLD phenotype was found in 155 patients (27·7%). The 511 patients with informative morphological and cytogenetic data were classified into three groups according to karyotype: favourable, intermediate and adverse risk groups (92, 375 and 44 patients respectively). Normal karyotype was the most frequent as a total, and among both the non‐TLD and TLD patients (164 patients 45·3% and 78 patients 52·7% respectively). All but one patient with AML/TLD was classified into the intermediate or adverse cytogenetic risk group. TLD phenotype was associated with lower remission rate and shorter overall survival but it did not influence disease‐free survival. Although we did not find any specific cytogenetic abnormalities for AML/TLD, the rarity of favourable karyotypes among TLD patients indicates the biological difference between AML/TLD and AML/non‐TLD.
Assessing diversity: immune reconstitution and T‐cell receptor BV spectratype analysis following stem cell transplantation
Authors: Peggs Karl S. , Verfuerth Stephanie , D'Sa Shirley , Yong Kwee , Mackinnon Stephen
Published: March 2003

Summary. T‐cell receptor (TCR) BV spectratyping provides information concerning immune reconstitution complementary to that derived from monoclonal antibody analysis of surface antigen expression. However, the most appropriate way to analyse and represent these data, the number of subfamilies that should be analysed, and the effects of CD4/8 ratio on observed diversity, remain unclear. A diversity scoring system was developed based on analysis of 11 cord blood (CB) and 12 normal adult BV spectratypes. CB subfamily spectratypes demonstrated minor deviations from a Gaussian pattern consistent with current knowledge about germline TCR rearrangements. Diversity scores were significantly lower in myeloma patients than normal adults (P < 0·001) and fell significantly following stem cell transplantation (P = 0·003). Subsequently increasing diversity was not detected by two previously described complexity scoring systems. The CD4/8 ratio was neither the major determinant of the absolute diversity score nor of the change in score for individual patients, suggesting that analysis of unselected mononuclear cells can provide information largely independent of CD4/8 ratios. There was a relatively low correlation between individual subfamily scores and overall diversity score. The novel and objective diversity scoring system described appears better able to document changes in spectratype patterns than previously described techniques and should aid the standardization of reporting.