bluuebunny/crossref_metadata_2025 huggingface dataset:
Factors affecting the outcome of allogeneic bone marrow transplantation for adult patients with refractory or relapsed acute leukaemia
Authors: Grigg Andrew P. , Szer Jeff , Beresford Jennifer , Dodds Anthony , Bradstock Ken , Durrant Simon , Schwarer Anthony P. , Hughes Timothy , Herrmann Richard , Gibson John , Arthur Chris , Matthews Jane
Published: March 2003

We evaluated the outcome of allogeneic bone marrow transplantation (BMT) for advanced acute myeloid leukaemia (AML) and acute lymphoblastic leukaemia (ALL) in 383 adult patients in nine Australian adult BMT centres between 1981 and 1997. The median overall survival for the group was 4.8 months, with an estimated 5‐year survival of 18%. 28% of patients died of transplant‐related toxicities within the first 100 d. Progressive disease was responsible for 48% of deaths. Multi‐factor analysis demonstrated that AML (v ALL), disease status (second complete remission [CR2] v others), age (< 40 years) and duration of prior first complete remission (CR1) (> 6 months) were pre‐transplant variables significantly associated with improved survival. Acute graft‐versus‐host disease (GVHD) of any grade reduced the rate of relapse in both AML and ALL, but only grades I–II were associated with improved survival. Both limited and extensive chronic GVHD were associated with increased survival. Only patients with AML in untreated first relapse or CR2, with a duration of CR1 > 6 months, or patients with T ALL, had a 5‐year survival > 20%. Transplants for AML in induction failure or pre‐B ALL in untreated first relapse or CR2 had an intermediate outcome, with 5‐year survival of 10–20%. A 5‐year survival of < 10% was observed for patients transplanted for ALL in induction failure or for pre‐B ALL or AML in refractory first relapse or beyond CR2. These results suggest that for most adult patients with advanced acute leukaemia an allograft offers only a small chance of cure.
X‐chromosome inactivation analysis of isolated Reed‐Sternberg cells in nodular sclerosing Hodgkin's disease
Authors: Chang Hsiao‐Wen , Chong Siew‐Meng , Peh Suat‐Cheng , Lee Szu‐Hee
Published: March 2003

In spite of several studies of immunoglobulin (Ig) gene rearrangements in whole Hodgkin's disease (HD) tissues and in isolated single Reed‐Sternberg (RS) cells, the issue of clonality of the RS cell in HD remains incompletely resolved. Analysis of X‐chromosome inactivation patterns (XCIPs) can be used to determine whether cell populations are clonal in origin. By PCR amplification of the human androgen receptor (HUMARA) loci using nested primers, we have studied XCIPs in six cases of HD of the nodular sclerosing (NS) subtype in which individual RS cells were isolated by micromanipulation from formalin‐fixed, paraffin‐embedded tissue sections immunostained with CD30 or CD15. In order to assess whether a clonal population of RS cells might be present in NSHD tissues, we compared the XCIPs obtained from whole NSHD tissues with those obtained from single RS cells harvested from the same tissues. Whole tissues from all six cases of NSHD showed balanced HUMARA allelic patterns, whereas an average of 83% (range 77–91%) of single RS cells from each of the six tissues expressed the same high‐ or low‐molecular weight allele, suggesting that a clonal population of RS cells was likely to be present in each case. These data are consistent with the presence of a clonal population of RS cells in NSHD tissues.
Immune complexes inhibit apoptosis of chronic lymphocytic leukaemia B cells
Authors: Gamberale Romina , Geffner Jorge R. , Trevani Analía , Cherñavsky Alejandra , Scolnik Mariano , Arrosagaray Guillermo , Sarmiento Marcela , Giordano Mirta
Published: March 2003

In the present study we examined the effect of immune complexes (IC) on the survival of chronic lymphocytic leukaemia (B‐CLL) B cells. Our results showed that either precipitating IC (pIC), Ab‐coated erythrocytes (E‐IgG) or heat‐aggregated IgG (aIgG) significantly inhibited spontaneous apoptosis of B‐CLL cells, as well as that induced by fludarabine, chlorambucil or dexamethasone. After depletion of T lymphocytes, monocytes and NK cells, incubation with IC was no longer able to delay B‐CLL cells apoptosis, suggesting that prevention of apoptosis depends on IC interaction with accessory leucocytes. The release of IFNγ by non‐malignant cells upon activation with IC was responsible, to some extent, for IC effects as shown by the fact that neutralizing anti‐IFNγ MoAb partially prevented their ability to inhibit B‐CLL cells apoptosis. The observation that treatment with IC resulted in increased expression of HLA‐DR on B‐CLL cells suggests that inhibition of apoptosis is associated with cellular activation.
Leukaemia inhibitory factor enhances tissue factor expression in human monocyte‐derived macrophages: a gp130‐mediated mechanism
Authors: Meisel Simcha R. , Shimon Ilan , Edgington Thomas S. , Melmed Shlomo , Cercek Bojan , Shah Prediman K.
Published: March 2003

Leukaemia inhibitory factor (LIF) and interleukin (IL)‐6 are members of a cytokine group that share a common signal transducer gp130 and induce pleiotropic biological effects in cells of diverse lineage. In monocytes, LIF facilitates differentiation, which may stimulate the biosynthesis of tissue factor (TF) that initiates the coagulation cascade. We tested the hypothesis that LIF would enhance TF expression in human monocyte‐derived macrophages (MDMs). Human peripheral blood mononuclear cells separated from whole blood by density centrifugation were allowed to differentiate into MDMs in primary culture, and were then exposed to LIF, IL‐6 and oncostatin M (OSM) for 24 h. LIF and IL‐6 receptors, and gp130 were demonstrated in MDMs by immunocytochemistry and RT‐PCR. TF procoagulant activity (TF‐PCA) was measured by recalcification clotting time and TF protein by Western blotting. The results show that both TF procoagulant activity and TF protein increased significantly in response to LIF over the concentration range of 1–100 nm (P < 0.03). Although OSM and IL‐6 tended to enhance TF expression by MDMs, the increase did not reach statistical significance. Anti‐LIF receptor and anti‐gp130 antibodies attenuated the effect of LIF on TF expression as assayed by both bioassay and flow‐cytometry. In conclusion, LIF increases TF‐PCA and TF protein in MDMs, and specific anti‐LIF receptor antibodies attenuate this effect. Thus, LIF may regulate by a gp130‐dependent pathway macrophage‐mediated procoagulant function in diverse pathological states involving inflammation and thrombosis and seems to serve as an important mediator at the interface between these processes.
Marked heterogeneity in protein levels and functional integrity of the thrombopoietin receptor c‐mpl in polycythaemia vera
Authors: Le Blanc  , Andersson  , Samuelsson
Published: March 2003

Polycythaemia vera (PV) is a myeloproliferative disorder (MPD) characterized by an increased production of mature blood cells. The underlying pathogenic mechanisms behind PV are largely unknown. Thrombopoietin (TPO) is the most important cytokine for stimulation of megakaryocyte growth and formation of functional platelets. Recently, it has been shown that the receptor for TPO, c‐mpl, is expressed on haematopoietic stem cells, and that TPO promotes the growth of these stem cells via binding to c‐mpl. Quantitative or qualitative abnormalities of c‐mpl function could thus theoretically play a role in the pathogenesis of different MPDs. Previous studies of the integrity of the c‐mpl system in PV have produced conflicting results. We therefore studied c‐mpl protein expression using immunoblot analysis in 15 PV patients and 10 healthy controls. Seven out of 15 PV patients (47%) exhibited similar c‐mpl protein levels to the controls, whereas eight out of 15 patients (53%) showed either markedly reduced or absent levels of c‐mpl. Five of the seven c‐mpl‐positive patients had only been treated by phlebotomy, whereas six out of eight c‐mpl‐negative patients were receiving treatment with hydroxyurea, anagrelide or α‐interferon. Disease duration tended to be slightly longer in c‐mpl‐negative patients compared with c‐mpl‐positive patients (mean = 55 vs. 43 months). Tyrosine phosphorylation of JAK‐2 in immunoprecipitates of platelets obtained after stimulation with TPO (100 and 1000 ng/ml) was normal in c‐mpl‐positive patients, whereas it could not be detected in c‐mpl‐negative patients. We therefore conclude that there exists a marked heterogeneity in c‐mpl protein levels and functional integrity in PV. However, it seems less likely that c‐mpl abnormalities per se are directly involved in the pathogenesis leading to the occurrence of PV, as c‐mpl levels were similar to those seen in healthy individuals in about half of the patients under study.
Increased soluble P‐selectin levels following deep venous thrombosis: cause or effect?
Authors: Blann Andrew D. , Noteboom Willy M. P. , Rosendaal Frits R.
Published: March 2003

Deep vein thrombosis (DVT) is associated with coagulation abnormalities, but evidence of excess platelet activity is scant. Soluble P‐selectin is a marker of platelet activity, with high levels being found in patients with thrombotic disease. We measured soluble P‐selectin by enzyme‐linked immunosorbent assay (ELISA) in plasma from 89 patients with objectively confirmed DVT and in 126 healthy age‐ and sex‐matched control subjects, and found higher levels in the patients (P = 0.011). Taking the risk of DVT with a level of soluble P‐selectin < 238 ng/ml to be 1, the relative risk of DVT with a soluble P‐selectin level >238 ng/ml was 2.1 (95% CI 1.2–3.6). These high levels may be a reflection of a generalized hypercoagulable state that, with factors such as the presence of persistent thrombin generation, could be responsible for excess platelet activation.
Intracellular accumulation of factor VIII induced by missense mutations Arg593→Cys and Asn618→Ser explains cross‐reacting material‐reduced haemophilia A
Authors: Roelse  , De Laaf  , Timmermans  , Peters  , Van Mourik  , Voorberg
Published: March 2003

Patients with cross‐reacting material (CRM)‐reduced haemophilia A exhibit reduced levels of factor VIII antigen. In this study, we determined the molecular basis of the genetic defect in the factor VIII gene induced by either the Arg593→Cys or the Asn618→Ser missense mutation, identified in two CRM‐reduced haemophilia A patients. We introduced either the Arg593→Cys or the Asn618→Ser mutation into a B‐domain‐deleted factor VIII cDNA and expressed the modified cDNAs in C127 cells. Reduced levels of factor VIII activity and factor VIII antigen in conditioned medium of transfected cells indicated that the secretion of both factor VIII variants was impaired. The ratio of factor VIII antigen present in cell extract to that in conditioned medium was 1.9 and 2.4 times higher for rFVIII‐R593C and rFVIII‐N618S, respectively, than for rFVIII. Metabolic labelling and immunoprecipitation of transfected cells revealed that rFVIII‐R593C and rFVIII‐N618S persisted somewhat longer inside the cell than factor rFVIII. Intracellular accumulation and subsequent degradation of factor VIII‐R593C and factor VIII‐N618S may explain the reduced levels of both factor VIII activity and antigen in plasma of mild haemophilia A patients with corresponding genetic defects.
Prevention of worsening of severe thrombocytopenia after red cell transfusions by the use of leucocyte‐depleted blood
Authors: Schuh  , Atoyebi  , Littlewood  , Hatton  , Bradburn  , Murphy
Published: March 2003

Platelet counts were measured before and after red cell transfusions in 30 patients with anaemia and severe thrombocytopenia resulting from haematological diseases. There was a mean reduction of 1.1 × 109/l (P = 0.43) in the platelet count after transfusions of 2–3 units of leucocyte‐depleted red cell concentrates (20 patients). However, there was a mean reduction of 2.7 × 109/l (P = 0.03), ≈ 10%, in the platelet count after transfusions of non‐leucocyte‐depleted red cell concentrates (10 patients). The findings suggest that the forthcoming introduction of universal leucocyte depletion of red cell concentrates will minimize the worsening of thrombocytopenia that occurs in severely thrombocytopenic patients receiving standard non‐leucocyte‐depleted red cell concentrates.
Lamivudine allows completion of chemotherapy in lymphoma patients with hepatitis B reactivation
Authors: Silvestri  , Ermacora  , Sperotto  , Patriarca  , Zaja  , Damiani  , Fanin  , Baccarani
Published: March 2003

Reactivation of hepatitis B virus in patients receiving chemotherapy for non‐Hodgkin's lymphoma (NHL) may give rise to hepatitis, hepatic failure and death, and prevent further chemotherapy. We report four patients with NHL in whom hepatitis flare‐up was observed after two (three patients) and six (one patient) cycles of chemotherapy. After spontaneous recovery, they were treated with Lamivudine (100 mg/day), which enabled completion of chemotherapy without further hepatitis B reactivation. In one patient, high‐dose chemotherapy and autologous stem cell transplantation was also performed. These data suggest a possible role for Lamivudine in preventing hepatitis B reactivation during chemotherapy administration to chronic carriers of the hepatitis B virus. Moreover, it enabled the completion of both standard and high‐dose chemotherapy in patients with previous hepatitis B reactivation.
Second allogeneic haematopoietic stem cell transplantation in relapsed acute and chronic leukaemias for patients who underwent a first allogeneic bone marrow transplantation: a survey of the Société Française de Greffe de Moelle (SFGM)
Authors: Michallet  , Tanguy  , Socié  , Thiébaut  , Belhabri  , Milpied  , Reiffers  , Kuentz  , Cahn  , Blaise  , Demeocq  , Jouet  , Michallet  , Ifrah  , Vilmer  , Molina  , Michel  , Lioure  , Cavazzana‐Calvo  , Pico  , Sadoun  , Guyotat  , Attal  , Curé  , Bordigoni  , Sutton  , Buzyn‐Veil  , Tilly  , Leporrier  , Fegueux  , Dreyfus  , Rio  , Lutz  , Vernant
Published: March 2003

Although recurrent malignancy is the most frequent indication for second stem cell transplantation (2nd SCT), there are few reports that include sufficiently large numbers of patients to enable prognostic factor analysis. This retrospective study includes 150 patients who underwent a 2nd SCT for relapsed acute myeloblastic leukaemia (n = 61), acute lymphoblastic leukaemia (n = 47) or chronic myeloid leukaemia (n = 42) after a first allogeneic transplant (including 26 T‐cell‐depleted). The median interval between the first transplant and relapse, and between relapse and second transplant was 17 months and 5 months respectively. After the 2nd SCT, engraftment occurred in 93% of cases, 32% of patients developed acute graft‐vs.‐host disease (GVHD)  grade II and 38% chronic GVHD. The 5‐year overall and disease‐free survival were 32 ± 8% and 30 ± 8%, respectively, with a risk of relapse of 44 ± 12% and a transplant‐related mortality of 45 ± 9%. In a multivariate analysis, five factors were associated with a better outcome after 2nd SCT: age < 16 years at second transplant; relapse occurring more than 12 months after the first transplant; transplantation from a female donor; absence of acute GVHD; and the occurrence of chronic GVHD. The best candidates for a second transplant are likely to be patients with acute leukaemia in remission before transplant, in whom the HLA‐identical donor was female and who relapsed more than 1 year after the first transplant.
Cell cycle distribution of cord blood‐derived haematopoietic progenitor cells and their recruitment into the S‐phase of the cell cycle
Authors: Lucotti  , Malabarba  , Rosti  , Bergamaschi  , Danova  , Invernizzi  , Pecci  , Ramajoli  , Perotti  , Torretta  , De Amici  , Salvaneschi  , Cazzola
Published: March 2003

The objective of this study was to evaluate the cycling status of cord blood (CB)‐derived colony‐forming cells (CFC) and long‐term culture‐initiating cells (LTC‐IC), and their recruitment into the S‐phase of the cell cycle. By using the cytosine arabinoside (Ara‐C) suicide approach, we found that only small proportions of both CFC and LTC‐IC were in the S‐phase of the cell cycle. These estimates were confirmed by flow cytometric DNA analysis, which showed that 96 ± 2% of CB‐derived CD34+ cells were in G0/G1 and only 1.6 ± 0.4% in the S‐phase. Staining of CD34+ cells with an antistatin monoclonal antibody, a marker of the G0 phase, indicated that among CD34+ cells with a flow cytometric DNA content typical of the G0/G1 phase 68 ± 7% of cells were in the G0 phase of the cell cycle. Incubation (24 h) with interleukin 3 (IL‐3), recombinant human stem cell factor (SCF) and granulocyte colony‐stimulating factor (G‐CSF) significantly increased the proportion of cells in the S‐phase for both CFC and LTC‐IC without inducing any loss in numbers. Flow cytometric DNA analysis also showed an increase in CD34+ cells in the S‐phase upon continuous exposure to these cytokines. Our findings indicate that: (i) very few CB‐derived CFC or LTC‐IC were in the S‐phase of the cell cycle; (ii) a substantial amount of CD34+ cells with a flow cytometric DNA content typical of the G0/G1 fraction was cycling, as found in the G1 phase of the cell cycle; and (iii) 24‐h incubation with IL‐3, SCF and G‐CSF could drive a proportion of progenitor cells into the S‐phase without reducing their number. These data might be useful for gene transfer protocols and the ex vivo expansion of CB‐derived progenitor cells.
Expression of HLA‐C‐specific natural killer cell receptors (CD158a and CD158b) on peripheral blood mononuclear cells after allogeneic bone marrow transplantation
Authors: Tanaka Junji , Mori Akio , Ohta Shuichi , Yamamoto Yasushi , Kobayashi Sumiko , Hashino Satoshi , Kobayashi Masanobu , Asaka Masahiro , Imamura Masahiro
Published: March 2003

We investigated the expression of natural killer cell receptors (NKRs) for HLA‐C on peripheral blood mononuclear cells (PBMCs) in 23 allogeneic bone marrow transplantation (allo‐BMT) patients to analyse the role of NKRs in alloresponse concerning graft‐versus‐host disease (GVHD). CD158a expression was low and there was little change in the expression after allo‐BMT. Also, there was no difference in the proportion of CD158a+/CD3− after allo‐BMT. In contrast, the proportion of CD158b+/CD3− cells, mainly NK cells, increased in the early stage (< 2 months) after allo‐BMT and then gradually decreased (3.3 ± 2.6% before BMT vs. 15.4 ± 8.6% in the early stage after BMT, 8.5 ± 4.9% during the period 3–6 months after BMT and 7.0 ± 3.0% > 6 months after BMT; P < 0.05). However, CD158b expression on CD3+ T cells increased 3 months after allo‐BMT (1.1 ± 1.1% before BMT vs. 5.1 ± 7.7% during the period 3–6 months after BMT and 3.0 ± 2.4% > 6 months after BMT, P < 0.05). The highest percentages of CD158 expression in patients without chronic GVHD (cGVHD) and those with cGVHD were compared. The percentage of CD158b+/CD3+ cells and also that of CD158b+/CD8+ cells were significantly increased in patients with cGVHD compared with those in patients without cGVHD (2.6 ± 2.0% vs. 8.0 ± 11.2% and 2.3 ± 1.5% vs. 8.3 ± 11.7% respectively; P < 0.05). The exact clinical relevance of these CD158b‐expressing cells is not clear. However, there is an interesting possibility that CD158b‐expressing cells play some role in the regulation of GVHD after allo‐BMT.
Flow cytometric and functional characterization of AC133+ cells from human umbilical cord blood
Authors: Pasino Mirella , Lanza Tiziana , Marotta Fernando , Scarso Lucia , De Biasio Pierangela , Amato Stefano , Corcione Anna , Pistoia Vito , Mori Pier Giorgio
Published: March 2003

AC133+ cells may represent an alternative source of transplantable haemopoietic progenitor cells to CD34+ cells. Here, we have addressed the characterization of umbilical cord blood (UCB) AC133+ cells and compared their immunophenotypic and functional features with those of UCB CD34+ cells. UCB AC133+ and CD34+ cell fractions were purified by magnetic cell sorting, analysed by flow cytometry, tested for their content in blast cell colony‐forming units (CFU‐Bl), erythroid and granulocyte–macrophage colony‐forming units before and after expansion in the presence of various haemopoietic growth factor combinations. Median AC133+ cell yield was 62·3%, and median AC133+ population purity was 97·9%. AC133+ cells were found to contain significantly more CFU‐Bl than CD34+ cells; furthermore, the replating efficiency, i.e. the number of CFU‐Bl capable of generating secondary colonies, was higher in the former than in the latter cells. Both AC133+ and CD34+ cells displayed an increased ability to give rise to committed progenitors after 7‐day expansion in liquid cultures. These data suggest that the AC133+ cell subset is a heterogeneous pool of immature and more differentiated cells that can be maintained and expanded in well‐defined culture conditions. In comparison with CD34+ cells, UCB AC133+ cells appear to contain a higher number of early haemopoietic progenitors.
Haemophagocytic syndrome associated with Plasmodium vivax infection
Authors: Aouba A. , Noguera M. E. , Clauvel J. P. , Quint L.
Published: March 2003

A 41‐year‐old woman was admitted with fever, splenomegaly and pancytopenia. High serum ferritin, hypertriglyceridaemia and bone marrow haemophagocytosis were consistent with a haemophagocytic syndrome. Trophozoites and gametocytes of Plasmodium vivax were identified on blood smear. Rapid recovery was observed after treatment with oral chloroquine.
Increased expression of non‐functional killer inhibitory receptor CD94 in CD8+ cells of myeloma patients
Authors: Besostri Barbara , Beggiato Eloise , Bianchi Alberto , Mariani Sara , Coscia Marta , Peola Silvia , Foglietta Myriam , Boccadoro Mario , Pileri Alessandro , Moretta Lorenzo , Massaia Massimo
Published: March 2003

Different MHC class I‐specific killer inhibitory receptors (KIRs) are expressed in vivo by a minor fraction of activated memory CD8+ cells. It has been postulated that KIRs may ‘fine‐tune’ specific responses by altering their threshold of activation by the TCR–CD3 complex. We have previously shown that, in multiple myeloma (MM) patients, a large fraction of peripheral blood CD8+ cells display the phenotype of chronically activated memory T cells (CD38+, HLA‐DR+, CD25−, CD45R0+, CD28−). We investigated the expression of KIRs on MM T cells and determined their possible influence on cytolytic responses elicited via the CD3–TCR complex. The expression of CD94, a molecule that is part of a heterodimeric KIR recognizing the non‐classical MHC surface HLA‐E molecule, was almost threefold higher in MM T cells than in age‐matched normal control subjects (P < 0·0001). CD94 expression was preferentially confined to CD8+ cells but not restricted to activated (HLA‐DR+) and/or memory (CD45R0+) T cells. Unlike normal T cells, in which CD94 is assembled with glycoproteins of the NKG2 family to form functional receptors with activating or inhibitory properties, most CD94+ MM T cells were devoid of both the NKG2‐A and NKG2‐C glycoproteins detected in the inhibitory or activating form respectively. CD94 blockade did not significantly affect either T‐cell proliferation or cytotoxic T‐lymphocyte generation induced by the myeloma‐derived cell lines NCI and RPMI 8226. Similarly, the cytolytic activity induced by direct anti‐CD3‐mediated targeting of MM T cells to FCR+ P815 target cells was unaffected by the addition of anti‐CD94 and/or anti‐NKG2‐A/C monoclonal antibodies (mAbs). These data indicate that the large majority of MM CD8+ cells do not express a functional CD94 receptor. Thus, their ability to ‘fine‐tune’ an appropriate immune response against tumour cells can be impaired.
Expression of T‐cell‐associated antigens in B‐cell non‐Hodgkin's lymphoma
Authors: Inaba Tohru , Shimazaki Chihiro , Sumikuma Toshiya , Okano Akira , Hatsuse Mayumi , Okamoto Akio , Takahashi Ryoichi , Ashihara Eishi , Hibi Shigeyoshi , Sudo Yoshikazu , Yamagata Noboru , Murakami Satoshi , Rin Kenhiro , Fujita Naohisa , Yoshimura Manabu , Nakagawa Masao
Published: March 2003

We performed the immunophenotyping of 101 patients with B‐cell non‐Hodgkin's lymphoma (B‐NHL) using two‐colour flow cytometry (FCM) and found that lymphoma cells coexpressed at least one kind of T‐cell‐associated antigen (T‐Ag; CD2, CD5, CD7) in 25 patients (24·8%). Among these three T‐Ags, CD5 was the most frequently expressed, in 21 patients (20·8%), followed by CD7, expressed in five patients (5·0%), and CD2, which was expressed in two patients (2·0%). Two kinds of T‐Ag were simultaneusly expressed in three patients (CD2/CD5, CD2/CD7, and CD5/CD7, each expressed in one patient). Concerning the expression pattern of T‐Ag, there were no significant differences between lymph nodes and extranodal organs in the three patients with T‐Ag‐positive B‐NHL (T‐Ag(+) B‐NHL) who were analysed. When comparing the clinical features between T‐Ag(+) B‐NHL and T‐Ag‐negative B‐NHL (T‐Ag(–) B‐NHL), extranodal involvement and higher International Prognostic Index (H and H.I.) were significantly frequent in the former subgroup (P = 0·0119 and P = 0·0302 respectively).
Activity of interleukin 6 in the differentiation of monocytes to macrophages and dendritic cells
Authors: Mitani Hidetsugu , Katayama Naoyuki , Araki Hiroto , Ohishi Kohshi , Kobayashi Kyoko , Suzuki Hirohito , Nishii Kazuhiro , Masuya Masahiro , Yasukawa Kiyoshi , Minami Nobuyuki , Shiku Hiroshi
Published: March 2003

Peripheral blood monocytes are common precursor cells of dendritic cells (DCs) and macrophages. We have searched for factors with the potential to regulate the differentiation of monocytes to DCs and macrophages. When CD14+ monocytes are cultured with granulocyte–macrophage colony‐stimulating factor (GM‐CSF) and interleukin (IL) 4, the CD14+CD1a− population, which consists of macrophages, was found in the serum‐containing cultures but not in the serum‐free cultures. Addition of IL‐6 receptor‐neutralizing monoclonal antibody (mAb) or gp130‐neutralizing mAb to the serum‐containing cultures resulted in a decreased population of CD14+CD1a− cells. An increase in the CD14+CD1a− population with reduction in CD14−CD1a+ DCs was observed with the addition of IL‐6 to cultures, whereas IL‐11, leukaemia inhibitory factor, oncostatin M or macrophage colony‐stimulating factor did not affect the differentiation of monocytes in the presence of GM‐CSF plus IL‐4. This effect of IL‐6 was blocked by tumour necrosis factor α (TNF‐α), lipopolysaccharide (LPS), IL‐1β, CD40 ligand (CD40L) and transforming growth factor β1 (TGF‐β1). Among these factors, TNF‐α was most potent in interfering with the action of IL‐6. These results suggest that IL‐6 inhibits the differentiation of monocytes to DCs by promoting their differentiation toward macrophages, which is modulated by factors such as TNF‐α, LPS, IL‐1β, CD40L and TGF‐β1.
Lymphocyte subset abnormalities in multitransfused HIV‐negative haemophilia A patients are not due to chronic hepatitis C virus infection
Authors: Meijer K. , Van Der Meer J. , Smit J. W. , Verspiek S. P. J. , Haagsma E. B. , Smid W. M.
Published: March 2003

Several abnormalities of immune parameters have been described in HIV‐negative haemophiliacs, including changes in numbers of T4 and T8 cells, T4/T8 ratio and numbers of activated T cells. To assess the contribution of hepatitis C to these abnormalities, we compared lymphocyte subsets in 20 HIV‐negative HCV‐positive haemophilia A patients and in 14 non‐haemophiliacs with chronic hepatitis C with those in 18 healthy controls. In haemophilia patients, higher numbers of T lymphocytes and activated T8 cells, lower numbers of B lymphocytes and a normal T4/T8 ratio were seen. These differences were either not demonstrated in hepatitis C controls or were less pronounced and in the opposite direction. We conclude that haemophilia A patients showed abnormalities in lymphocyte subsets that could not be attributed to chronic hepatitis C.
Captopril inhibitsin vitroandin vivothe proliferation of primitive haematopoietic cells induced into cell cycle by cytotoxic drug administration or irradiation but has no effect on myeloid leukaemia cell proliferation
Authors: Chisi John E. , Briscoe Cecilia V. , Ezan Eric , Genet Roger , Riches Andrew C. , Wdzieczak‐Bakala Joanna
Published: March 2003

Angiotensin I‐converting enzyme (ACE) has been shown to be involved in the catabolism of the tetrapeptide acetyl‐Ser–Asp–Lys–Pro (AcSDKP). As AcSDKP is a physiological inhibitor of haematopoietic stem cell proliferation, we investigated thein vitroandin vivoeffects of captopril, one of the specific inhibitors of ACE, on the proliferation of primitive haematopoietic cells. Regenerating bone marrow cells obtained from mice given one injection of cytosine arabinoside (100 mg/kg) as well as SA2 myeloid leukaemia cells were incubatedin vitrofor 24 h with 10−6 mcaptopril. Captopril significantly reduced the proportion of high proliferative potential colony‐forming cells (HPP‐CFC‐1) in S‐phase, whereas it had no effect on the proportion of SA2 leukaemic colony‐forming cells in S‐phase. When givenin vivoto mice 1 h after 2 Gy γ‐irradiation or cytosine arabinoside (AraC) injection, captopril (100 mg/kg) was shown to prevent HPP‐CFC‐1 entry into S‐phase induced by these cytotoxic treatments. The observed effects correlated with a reduction in ACE degradative activity and an increase in the level of endogenous AcSDKP both in the supernatants of captopril‐treated bone marrow cells and in plasma of treated animals. The present findings suggest that AcSDKP might mediate the observedin vitroandin vivoinhibitory effects of captopril on primitive haematopoietic cell proliferation.
Role of A and B blood group antigens in the expression of adhesive activity of von Willebrand factor
Authors: Sarode Ravindra , Goldstein Jack , Sussman Ira I. , Nagel Ronald L. , Tsai Han‐Mou
Published: March 2003

ABO (H) blood group antigens are covalently linked to the oligosaccharide side‐chains of von Willebrand factor (VWF). In this study, we investigated the role of the A and B antigens in the expression of VWF adhesive activity. VWF of type A, B or O was purified from fresh frozen plasma. Presence of A or B antigen on the VWF was confirmed by enzyme‐linked immunosorbent assay (ELISA) and by immunoblotting with monoclonal anti‐A or anti‐B. The A or B antigen was also detected in the 48/52‐kDa fragment of the respective VWF after trypsin digestion. Removal of A antigen with α‐N‐acetylgalactosaminidase or B antigen with α‐galactosidase did not affect its multimer size or antigenic level, but decreased the ristocetin cofactor (RCoF) activity of the respective VWF by 33–39% (P < 0·01–0·002). Removal of A or B antigen from VWF did not affect the binding of the VWF to immobilized type III collagen. A and B antigens were not detected in platelet VWF. These results indicate that AB structures play a role in platelet aggregating activity of VWF.
Activated protein C inhibits lipopolysaccharide‐induced nuclear translocation of nuclear factor κB (NF‐κB) and tumour necrosis factor α (TNF‐α) production in the THP‐1 monocytic cell line
Authors: White B. , Schmidt M. , Murphy C. , Livingstone W. , O'Toole D. , Lawler M. , O'Neill L. , Kelleher D. , Schwarz H. P. , Smith O. P.
Published: March 2003

Activated protein C (APC) protects against sepsis in animal models and inhibits the lipopolysacharide (LPS)‐induced elaboration of proinflammatory cytokines from monocytes. The molecular mechanism responsible for this property is unknown. We assessed the effect of APC on LPS‐induced tumour necrosis factor α (TNF‐α) production and on the activation of the central proinflammatory transcription factor nuclear factor‐κB (NF‐κB) in a THP‐1 cell line. Cells were preincubated with varying concentrations of APC (200 µg/ml, 100 µg/ml and 20 µg/ml) before addition of LPS (100 ng/ml and 10 µg/ml). APC inhibited LPS‐induced production of TNF‐α both in the presence and absence of fetal calf serum (FCS), although the effect was less marked with 10% FCS. APC also inhibited LPS‐induced activation of NF‐κB, with APC (200 µg/ml) abolishing the effect of LPS (100 ng/ml). The ability of APC to inhibit LPS‐induced translocation of NF‐κB is likely to be a significant event given the critical role of the latter in the host inflammatory response.
Treatment of chronic myelomonocytic leukaemia by allogeneic marrow transplantation
Authors: Zang Dae Young , Deeg H. Joachim , Gooley Ted , Anderson Jeanne E. , Anasetti Claudio , Sanders Jean , Myerson David , Storb Rainer , Appelbaum Fred
Published: March 2003

We evaluated the outcome of allogeneic bone marrow transplantation (BMT) in 21 patients with chronic myelomonocytic leukaemia (CMML) who were treated at the Fred Hutchinson Cancer Research Center between 1990 and 1998. There were 11 male and 10 female patients with a median age of 47·4 years (range 1·0–62·9). Patients were conditioned either with total body irradiation (TBI) and chemotherapy, with or without antithymocyte globulin (n = 19), or with chemotherapy alone (n = 2). The marrow donor was an HLA‐identical sibling in 12 patients, an HLA‐non‐identical related donor in three patients and an unrelated volunteer donor in six patients. All evaluable patients achieved sustained engraftment. Fifteen patients developed grades II–IV acute graft‐versus‐host disease (GVHD). Nine patients (43·0%) are surviving disease free at 0·7–8·1 years (median 6·9) after transplantation. Five patients relapsed 75–660 d after transplant and all died. Five patients died with organ failure and two died with GVHD and associated infections. The Kaplan–Meier estimates of disease‐free survival and relapse at 3 years were 39% and 25% respectively. The probability of survival was improved in patients with shorter disease duration compared with those with a long interval from diagnosis to BMT. Thus, as with other myeloproliferative diseases or myelodysplastic syndromes, BMT offers curative therapy for a proportion of patients with CMML. We suggest that patients with CMML who have a suitable donor should be considered for transplantation, probably early in their disease course. However, it will be important to develop new regimens with enhanced antileukaemic efficacy without further increasing regimen‐related toxicity and mortality.
Transfusion‐associated graft vs. host disease in a patient with high‐grade B‐cell lymphoma. Should cellular products for patients with non‐Hodgkin's lymphoma be irradiated?
Authors: Gelly K. J. , Kerr R. , Rawlinson S. , Norris A. , Bowen D. T.
Published: March 2003

We present the case of a 64‐year‐old man who died from transfusion‐associated graft vs. host disease (TA‐GVHD) having been treated 2 years earlier for a high‐grade, non‐Hodgkin's lymphoma (NHL). We suggest that he was at increased risk of developing TA‐GVHD as a result of the NHL and its subsequent treatment, and propose that patients with NHL should be added to those ‘at risk’ groups who receive irradiated cellular blood components.
Effect of nitric oxide and nitric oxide donors on red blood cell oxygen transport
Authors: Hrinczenko Borys W. , Alayash Abdu I. , Wink David A. , Gladwin Mark T. , Rodgers Griffin P. , Schechter Alan N.
Published: March 2003

A mechanism has been proposed in which nitric oxide (NO) may bind to cysteine β93 and be transported by haemoglobin from the lungs to the tissues and modify vascular tone. In addition, it has been reported that treatment of sickle cell anaemia blood with 80 p.p.m. NO gas in air shifts the oxygen affinity, as measured by P50to the left. We exposed normal and sickle cell anaemia blood to 80 p.p.m. NO in air for 1 h in vitro and found no change in P50 of either normal or sickle cell blood. In addition, we exposed normal and sickle cell blood in buffer to aqueous NO (NO gas dissolved in buffer) at varying concentrations and found that the induced left shift in P50 correlates strongly and linearly with methaemoglobin formation. We also treated normal and sickle cell blood with other nitric oxide donors, such as sodium 2‐(N,N‐diethylamino)‐diazenolate‐2‐oxide (DEANO), S‐nitrosocysteine (CysNO) and sodium trioxodinitrate (OXINO, or Angeli's salt). In all cases, we found a dose‐dependent increase in methaemoglobin that was strongly correlated with the dose‐dependent P50 reduction. Our data do not support the report that low NO concentrations can selectively increase the oxygen affinity of sickle cell blood without affecting methaemoglobin levels significantly. NO, however, may have benefit in sickle cell disease by other mechanisms.
Cytogenetic and molecular evidence of marrow involvement in extramedullary acute myeloid leukaemia
Authors: Lillington Debra M. , Jaju Rina J. , Shankar A. G. , Neat Michael , Kearney Lyndal , Young Bryan D. , Saha Vaskar
Published: March 2003

A diagnosis of granulocytic sarcoma was made in a 2‐year‐old child based on the detection of myelomonocytic blasts in tissue obtained from a subcutaneous nodule with no evidence of concomitant disease in the bone marrow. The child responded to systemic chemotherapy and is in remission 3 years later. An identical clone with an in frame fusion of the MLL and AF10 genes was identified from both tissue and bone marrow samples. The generation of an in frame MLL–AF10 fusion requires complex intra‐ and interchromosomal exchanges between chromosomes 10 and 11. In this case, an intrachromosomal rearrangement of chromosome 5 was also observed. This case illustrates the presence of systemic disease in extramedullary leukaemia, its response to systemic rather than topical therapy and suggests that the events leading to chromosomal translocations in leukaemia may be part of a generalized intracellular event.
Cyclin D1 and E2F‐1 immunoreactivity in bone marrow biopsy specimens of multiple myeloma: relationship to proliferative activity, cytogenetic abnormalities and DNA ploidy
Authors: Wilson CarlA. S. , Butch AnthonY. W. , Lai Raymond , Medeiros L. Jeffrey , Sawyer JeffreY. R. , Barlogie Bart , McCourty Althea , Kelly Kathy , Brynes RusselL. K.
Published: March 2003

Cyclin D1, encoded by the CCND1 gene, is immunohistochemically detectable in up to one‐third of cases of multiple myeloma (MM). To examine the mechanism of cyclin D1 overexpression, we compared cyclin D1 immunoreactivity with the results of conventional cytogenetics to determine if the t(11;14)(q13;q32) or other abnormalities of 11q11–14 explained cyclin D1 overexpression. Karyotypic abnormalities were found in 45 out of 67 (67%) MM cases; the t(11;14) was present in seven cases (10%). Additional 11q11–14 abnormalities were not identified. The t(11;14) correlated with cyclin D1 upregulation in low to intermediately proliferative MM, but was not present in highly proliferative tumours (assessed using bromodeoxyuridine labelling index). Cyclin D1 indirectly activates the transcription factor E2F‐1. In the bone marrow biopsy specimens of MM cases, E2F‐1 was concurrently expressed with cyclin D1 (P = 0·001), indicating that cyclin D1 is functional. However, as neither E2F‐1 nor cyclin D1 expression correlated with proliferative activity, the speculation that t(11;14) upregulates the CCND1 gene to induce higher proliferation and possibly more aggressive disease is not supported. We conclude that in low to intermediately proliferative MM cases, cyclin D1 is probably upregulated by t(11;14), but an alternative mechanism is more probable in highly proliferative MM.
Transient thrombopoietin peak after liver transplantation for end‐stage liver disease
Authors: Faeh M. , Hauser S. P. , Nydegger U. E.
Published: March 2003

Knowledge about synthesis of thrombopoietin (TPO) by human liver cells is now well established and makes the study of TPO serum levels and interdependency with platelet concentrations important before and after liver transplantation. We followed these variables in 14 patients suffering from primary biliary cirrhosis (group A), 10 with hepatitis B (group B) and nine with hepatitis C (group C) infections, as well as 11 patients suffering from alcoholic cirrhosis (group D). In the pretransplant serum samples, TPO levels and platelet counts revealed median values (range) of 112·75 pg/ml (5·0–349·3) in group A, 67·8 pg/ml (14·8–249·9) in B, 68·6 pg/ml (31·4–147·3) in C and 57·9 pg/ml (25·2–264·2) in D for TPO, and 138 × 109/l (36–243) in A, 48 × 109/l (22–129) in B, 105 × 109/l (32–176) in C and 109 × 109/l (33–285) in D for platelets, the latter levels being abnormally reduced. Within 2–3 d of transplantation, the TPO levels started to increase to transient peaks of mostly 5–10 times baseline, which were followed by continuous correction of thrombocytopenia. Splenomegaly was identified to be an important co‐factor of thrombocytopenia in groups A and D. We conclude that decreased TPO production in patients with end‐stage liver diseases is reverted by orthotopic liver transplantation.
Interleukin 12 induces activation of fibrinolysis and coagulation in humans
Authors: Portielje Johanna E. A. , Kruit Wim H. J. , Eerenberg Anke J. M. , Schuler Martin , Sparreboom Alex , Lamers Cor H. J. , Bolhuis Reinder L. H. , Stoter Gerrit , Huber Christoph , Hack C. Erik
Published: March 2003

Interleukin 12 (IL‐12) has potential efficacy in malignant, infectious and allergic diseases. Its side‐effects include activation of coagulation and fibrinolysis, as documented in chimpanzees. We assessed the coagulative and fibrinolytic response in 18 patients with renal cell carcinoma after subcutaneous injection of 0·5 μg/kg recombinant human IL‐12. IL‐12 induced a fibrinolytic response in 17 patients (94%): plasmin‐α2–anti‐plasmin complexes (PAPc) increased from 11·8 ± 6·6 nmol/l (mean ± SD) to a maximum of 18·8 ± 7·4 nmol/l at 72 h. Baseline levels of tissue plasminogen activator (tPA) and plasminogen‐activator inhibitor‐I (PAI) were elevated in eight and 14 patients respectively. tPA increased from 12·6 ± 5·2 ng/ml to a maximum of 19·0 ± 6·7 ng/ml at 72 h. PAI decreased from 111 ± 69 ng/ml to a minimum of 65 ± 53 ng/ml at 8 h, thereafter remaining below baseline. Elevation of PAPc correlated with elevation of tPA and reduction of PAI. A coagulative response occurred in nine patients (50%): thrombin–anti‐thrombin III complexes increased from 29 ± 53 ng/ml to a maximum of 460 ± 322 ng/ml at 12 h. Patients with and without a coagulative response had similar levels of recombinant human IL‐12, interferon‐γ or tumour necrosis factor‐α. We conclude that IL‐12 can activate both fibrinolysis and coagulation in a significant proportion of patients with cancer. The time‐frame and sequence of these activation processes differ from those known for other cytokines.
Reduced memory B‐cell populations in boys with B‐cell dysfunction after bone marrow transplantation for X‐linked severe combined immunodeficiency
Authors: Ting S. S. , Tangye S. G. , Wood J. , Ffrench R. A. , Ziegler J. B.
Published: March 2003

X‐linked severe combined immunodeficiency (XSCID) is a lethal disease resulting in death in infancy. In many instances, haploidentical bone marrow transplantation (BMT) offers reconstitution of T‐cell immunity alone, with residual hypogammaglobulinaemia. The exact nature of B‐cell dysfunction in these patients is unclear, although differentiation arrest of the B cells is a potential explanation. To ascertain the differentiation status of peripheral blood B lymphocytes from XSCID patients after BMT, the surface expression of CD19, CD10, CD34, CD5, serum immunoglogulin (sIg)M, sIgD, sIgG and CD27 on these B cells was investigated using three‐colour flow cytometry. CD27 is a marker of memory B cells. Populations of CD19+IgM−D− B cells, CD19+IgM‐only, CD19+IgG+CD27+ and CD19+IgM+ CD27+ B cells were found to be diminished in the XSCID patients after BMT with persistent hypogammaglobulinaemia, compared with both post‐BMT patients with B‐cell function and age‐matched normal controls. This indicated the lack of CD19+IgM−D− B cells, which represent Ig isotype‐switched B cells, as well as CD19+IgM‐only and CD19+IgG+CD27+ or CD19+IgM+CD27+ memory B‐cell populations. Interaction between CD27 and its ligand CD70 has been shown to induce IgG and IgM production by CD27+ B cells. Therefore, the lack of CD27/70 interaction is a probable explanation for the hypogammaglobulinaemia in these patients after BMT.
Myristoylated alanine‐rich C kinase substrate phosphorylation is involved in thrombin‐induced serotonin release from platelets
Authors: Elzagallaai A. , Rosé S. D. , Brandan N. C. , Trifaró J.‐M.
Published: March 2003

Stimulation of platelets by thrombin induces protein kinase C (PKC) activation, phosphorylation of pleckstrin, aggregation and serotonin release. Here, we demonstrate that, in human platelets, thrombin stimulation also induced phosphorylation of the myristoylated alanine‐rich C kinase substrate (MARCKS) and serotonin release in intact and digitonin‐permeabilized platelets. MARCKS is known to bind actin and cross‐link actin filaments, and this is inhibited by PKC‐evoked MARCKS phosphorylation. MARCKS phosphorylation and serotonin release in response to increasing concentrations of thrombin have a similar EC50 and time course and, in permeabilized platelets, peptide MPSD, with an amino acid sequence corresponding to the phosphorylation site domain of MARCKS, blocked both responses. However, pleckstrin and myosin light chain phosphorylations were not modified. Ala‐MPSD, in which the four serine residues of MPSD were substituted by alanines was ineffective. The results suggest a role for MARCKS in platelet secretion. The fact that pleckstrin phosphorylation has a different time course and was not modified in the presence of MPSD when MARCKS phosphorylation and serotonin release were inhibited would suggest either that pleckstrin phosphorylation is unrelated to secretion or that it might only be involved upstream in the events leading to secretion.
Characterization and localization of expression of an erythropoietin‐induced gene, ERIC‐1/TACC3, identified in erythroid precursor cells
Authors: McKeveney P. J. , Hodges V. M. , Mullan R. N. , Maxwell P. , Simpson D. , Thompson A. , Winter P. C. , Lappin T. R. J. , Maxwell A. P.
Published: March 2003

Gene expression profiles during erythropoietin (Epo)‐induced differentiation of erythroid progenitor cells derived from the Friend virus anaemia (FVA) and phenylhydrazine (PHZ) murine models have been examined using differential display polymerase chain reaction (PCR). Ten cDNA fragments upregulated by Epo were isolated. The ribonuclease protection assay confirmed differential expression between Epo‐stimulated and Epo‐deprived cells for one of these, provisionally named ERIC‐1. Sequencing of the full‐length cDNA predicted a protein of 558 amino acids, 17 amino acids longer than mTACC3, the third member of a novel family of proteins that contain a coiled‐coil domain. The human homologue, cloned using rapid amplification of cDNA ends (RACE)‐PCR, encodes a larger protein of 838 amino acids that is identical to hTACC3. In addition to erythroid precursor cells, ERIC‐1/TACC3 is expressed at high levels in the testes, at moderate levels in the thymus and peripheral leucocytes, and at lower levels in the spleen and intestinal tissue. Immunohistochemical analysis using an antibody to a GST fusion product of the C‐terminus of hERIC‐1/TACC3 revealed that it is localized to Sertoli cells in the human testes. Confocal microscopy demonstrated hERIC‐1/TACC3 protein concentrated in the perinuclear vesicles of dermal microvascular endothelial cells. Although ERIC‐1/TACC3 is expressed in a wide range of tissues, its upregulation by Epo in erythroid progenitors implies that it has a role in terminal erythropoiesis.