bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of interleukin‐4 production and cytokine‐induced growth potential in peripheral T‐cell non‐Hodgkin's lymphomas
Authors: Raziuddin Syed , Sheikha Anwar , Abu‐Eshy Saeed , Al‐Janadi Mansour
Published: March 2003

The malignant cells in tumour tissues produce cytokines/growth factors that may influence tumour growth, tumour immunogenicity and host immune response. We demonstrate that lymph node cell (LNC) purified neoplastic T cells from CD4+ peripheral T‐cell lymphoma (CD4+ PTCL) and CD8+ PTCL spontaneously, and after stimulation with anti‐CD3, secreted high amounts of interleukin‐4 (IL‐4) as compared to LNC‐purified CD4+ and CD8+ non‐malignant T cells. Furthermore, IL‐4 was observed to be the most potent cytokine that induced in vitro proliferation and growth of the malignant T cells. Moreover, malignant T‐cell‐derived IL‐4 secretion was augmented by exogeneous recombinant human interferon‐gamma (IFN‐γ) and was profoundly inhibited by IL‐2. Because IL‐4 was shown to be a locally active cytokine with a wide range of immunoregulatory properties, regulation of IL‐4 production by IFN‐γ and IL‐2 in malignant T cells may be one of the important parameters to be assessed in the design of anticancer‐specific immunotherapy. In summary, we report that malignant T cells produce IL‐4, a type 2 cytokine (Th2 cell response) that acts as a growth factor and which may play a critical role in PTCL disease mechanism.
Correlation of cytogenetic, molecular cytogenetic, and clinical findings in 59 patients with ANLL or MDS and abnormalities of the short arm of chromosome 12
Authors: Streubel Berthold , Sauerland Cristina , Heil Gerhard , Freund Mathias , Bartels Heinrich , Lengfelder Eva , Wandt Hannes , Ludwig Wolf‐Dieter , Nowotny Hadwiga , Baldus Michael , Grothaus‐Pinke Bernward , Büchner Thomas , Fonatsch Christa
Published: March 2003

Abnormalities of the short arm of chromosome 12 (12p) are found in about 5% of acute nonlymphocytic leukaemias (ANLL) and myelodysplastic syndromes (MDS). They are described to be characteristic of secondary leukaemias, especially after prior mutagenic exposure, and to be associated with a poor prognosis. In our series of 59 patients with 12p abnormalities and ANLL or MDS, exposure to genotoxic agents was proven only in five patients, but in 13/44 patients ANLL evolved from an MDS. Patients with a small deletion del(12)(p11.2p13) having a mild clinical course were distinguished from those with a large del(12)(p11.2), additional chromosomal anomalies, and a poor clinical course. Among the 31 patients with translocations or dicentric chromosomes involving 12p, a group of eight with t/dic(12;13) was the most frequent and was associated with a poor prognosis. The clinical outcome was adverse in the majority of patients with complex karyotype abnormalities, but in some patients a milder clinical course seems likely. A new, hitherto undescribed, abnormality in an MDS case with a duplication dup(12)(p11.2p13) was the amplification of the signal of the yeast artificial chromosome (YAC) clone 964c10 (D12S736). In 38 cases with deletions or unbalanced translocations/dicentrics one YAC signal was lost. Five patients with balanced translocations demonstrated breakpoints within the YAC, containing the ETV6 (TEL) gene. The breakpoints were telomeric to the YAC 964c10 in seven cases and centromeric in one patient.
Stimulation of thrombocytopoiesis decreases platelet β2 but not β1 or β3 integrins
Authors: Guo Jane , Piguet Pierre F.
Published: March 2003

The expression of CD29, CD61, CD18 and CD11a on platelets was examined by flow cytometry in mice treated with leukaemia inhibitory factor (LIF) or megakaryocyte growth and development factor (PEG‐rHuMGDF or mpl‐ligand). Treatment for 7–14 d with PEG‐rHuMGDF or LIF increased the number of platelets in peripheral blood from 0.9 up to <2.0×106/μl. These treatments decreased the expression of CD11a and CD18, whereas that of CD29 or CD61 was not markedly changed. Study after various doses or times of PEG‐rHuMGDF administration indicated that a decrease of CD18 expression occurred when platelet counts started to rise. Platelet RNA content was increased in mice treated with PEG‐rHuMGDF but double staining indicated that expression of CD18 was not correlated with RNA content. To evaluate integrin expression as a function of time in circulation, platelets were biotinylated in vivo. In normal or PEG‐rHuMGDF‐treated mice, the expression of CD29 or CD61 did not change, whereas that of CD18 decreased significantly as a function of time in circulation. These findings indicate, firstly, that stimulation of thrombocytopoiesis leads to the release of platelets with a low content of β2 integrin and, secondly, that this integrin is also selectively lost while in the circulation.
CD3−CD4−CD8−CD56−CD19−CD14− cells and CD3+CD8 dull‐positive cells produce IL‐4 in AIDS patients
Authors: Honda Junichi , Okubo Yasumitsu , Tanaka Ken , Kusaba Mikako , Kumagai Mutsuko , Saruwatari Naoko , Oizumi Kotaro
Published: March 2003

We used flow cytometry to identify the presence of intracellular cytokines (cytoflow) and analyse the production of IL‐4 in peripheral blood from AIDS patients who have practically no CD4+ T cells. We found that IL‐4 was produced by CD3−CD4−CD8−CD56−CD19−CD14− cells and CD3+CD8 dull‐positive cells in AIDS patients. Moreover, CD3−CD4− CD8−CD56−CD19−CD14− cells had helper activity for immunoglobulin synthesis. These findings indicate that instead of CD4+ T helper cells, C3−CD4−CD8−CD56− CD19−CD14− cells and CD3+CD8 dull‐positive cells may be an important source of IL‐4 in a variety of immune responses for AIDS patients.
A study to determine whether trisomy 8, deleted 9q and trisomy 22 are markers of cryptic rearrangements of PML/RARα, AML1/ETO and CBFB/MYH11 respectively in acute myeloid leukaemia
Authors: Langabeer  , Grimwade  , Walker  , Rogers  , Burnett  , Goldstone  , Linch
Published: March 2003

Acute myeloid leukaemia (AML) patients with either a t(15;17), t(8;21) or inv(16) at diagnosis have ‘good‐risk’ disease with a favourable response to therapy and improved survival. Detection of cryptic fusion genes created by these translocations has been reported where there is no cytogenetic evidence of the corresponding abnormality. It is likely that these cases share the same favourable prognosis. Secondary cytogenetic changes commonly associated with these rearrangements are +8 with t(15;17), del(9q) with t(8;21) and +22 with inv(16). These secondary abnormalities are also observed alone, raising the possibility that they may be markers of underlying cryptic rearrangements. In order to determine the frequency of these rearrangements in AML cases with +8, del(9q) or +22 we have performed an analysis of 63 such patients in whom there was no evidence of a t(15;17), t(8;21) or inv(16) by cytogenetics. No disease‐related fusion transcripts were identified, indicating that the secondary changes are rarely markers for cryptic rearrangements.
Prevention of central venous catheter associated thrombosis using minidose warfarin in patients with haematological malignancies
Authors: Boraks Paul , Seale James , Price Jane , Bass Gilda , Ethell Mark , Keeling David , Mahendra Prem , Baglin Trevor , Marcus Robert
Published: March 2003

Thrombosis is a well‐recognized complication following insertion of central venous catheters and is associated with significant morbidity. In an attempt to reduce line‐associated thrombosis, 108 consecutive patients with haematological malignancies were commenced on prophylactic ‘minidose’ warfarin, 1 mg/d, at the time of line insertion. This group of patients were compared with a historic group of 115 consecutive patients who had not received warfarin. Clinically‐suspected venous thrombosis was confirmed by Doppler ultrasound or venography. Patients taking prophylactic warfarin had their prothrombin time measured three times per week with the aim of maintaining an INR < 1.6. Five (5%) of the 108 patients who received minidose warfarin developed a thrombosis, at a median of 72 d (range 5–166) from the time of catheter insertion. In the 115 patients who were not anticoagulated 15 (13%) developed a catheter‐associated thrombosis at a median of 16 d (range 1–35). There was a significant reduction in line‐associated thrombosis in patients receiving warfarin (P = 0.03). These data suggest that minidose warfarin reduces the incidence of central venous catheter related thrombosis in patients with haematological malignancies.
Immunophenotypic and clinical features of T‐cell receptor γδ+ T‐lineage acute lymphoblastic leukaemia
Authors: Schott Gisela , Sperling Christian , Schrappe Martin , Ratei Richard , Martin Mathilde , Meyer Ulrike , Riehm Hansjörg , Ludwig Wolf‐Dieter
Published: March 2003

The immunophenotypic and clinical features of TCR‐γδ+ T‐lineage acute lymphoblastic leukaemia (T‐ALL) were prospectively analysed in 52 children with membrane CD3+ T‐ALL. We observed a relatively high incidence of TCR‐γδ+ T‐ALL (26/52 patients). Leukaemic blasts from 22 children demonstrated TCR‐αβ positivity, and simultaneous expression of the TCR‐β and ‐δ chain was found in four children. Clinical and haematological features of TCR‐αβ and γδ+ T‐ALL did not differ significantly, except that haemoglobin levels were significantly lower in TCR‐γδ+ cases. Event‐free survival at 4 years was significantly better in TCR‐γδ+ compared with TCR‐αβ+ T‐ALL, but expression of TCR molecules did not emerge as an independent prognostic factor in multivariate analysis.
A randomized trial of maintenance interferon following high‐dose chemotherapy in multiple myeloma: long‐term follow‐up results
Authors: Cunningham Dadvid , Powles Ray , Malpas James , Raje Noopur , Milan Sarah , Viner Christine , Montes Anna , Hickish Tamas , Nicolson Marianne , Johnson Paul , Treleaven Jennifer , Raymond Julian , Gore Martin
Published: March 2003

High‐dose chemotherapy (melphalan) with autologous marrow stem cell support (AMSCS) results in high response rates in multiple myeloma (MM), with up to 50% of patients achieving complete remission. However, these remissions are generally not durable. As the cytokine interferon alpha has been shown to prolong partial response following conventional chemotherapy, this trial was conducted to evaluate its role following high‐dose chemotherapy. 85 patients were randomly assigned to maintenance treatment with interferon alpha, 3 × 106 units/m2 subcutaneously three times weekly until relapse or no further treatment following recovery from high‐dose chemotherapy (melphalan 140–200 mg/m2 or busulphan 16 mg/kg) combined with AMSCS. At 5.8 years following the accrual of the last patient in this trial, 38 patients had died, 17 in the interferon arm and 21 in the control arm. The median progression‐free survival (PFS) in the 42 patients randomized to interferon alpha was 46 months versus 27 months in the controls. Both overall survival and PFS, which were highly significant at median follow‐up of 52 months, have now ceased to be significant, because most patients have ultimately succumbed to their disease. Interferon was tolerated by the majority of patients with very good compliance. Toxicity consisted mainly of flu‐like symptoms and malaise which were usually self‐limiting. The results of such a pilot study should be carefully interpreted and the benefits of interferon should be confirmed in larger multi‐centre studies in the setting of minimal residual disease following autologous transplantation.
IgG1‐κ biclonal gammopathy associated with multiple myeloma suggests a regulatory mechanism
Authors: Pizzolato  , Bragantini  , Bresciani  , Pavlovsky  , Chuba  , Vidal  , Rostagno  , Ghiso
Published: March 2003

Multiclonal gammopathies associated with multiple myeloma may result either from a neoplastic transformation of a cell clone undergoing immunoglobulin class switching or from independent transforming events yielding proliferation of unrelated plasma cell clones. The simultaneous presence of more than one neoplastic clone may possess regulatory implications in terms of cell proliferation, clonal expansion, secretion of M‐components or response to chemotherapy. We report a patient, diagnosed with multiple myeloma stage IIIa, who presented with two well‐defined homogenous IgG1‐κ components in the serum (designated WER‐1 and WER‐2) with striking differences in their plasma concentration and response to the classic melphalan/prednisone treatment. Immunochemical characterization and amino terminal sequence analysis of both the heavy and light chains of each M‐component undoubtedly determined their biclonal origin. WER‐1 was identified as IgG1(VHII)‐κI while WER‐2 was classified as IgG1(VHIII)‐κIII. The plateau phase was characterized by very low or undetectable levels of WER‐2, a high, almost constant, concentration of WER‐1 and the absence of Bence Jones proteinuria, whereas these parameters were completely reversed during the escape phase with levels resembling those observed at the time of diagnosis. The statistically significant negative correlation between the biclonal components and the different susceptibility to the treatment clearly suggests regulatory interactions between the clones WER‐1 and WER‐2.
Mutation analysis of CD95 (APO‐1/Fas) in childhood B‐lineage acute lymphoblastic leukaemia
Authors: Beltinger Christian , Böhler Thomas , Karawajew Leonid , Ludwig Wolf‐Dieter , Schrappe Martin , Debatin Klaus‐Michael
Published: March 2003

The CD95 system plays an important role in lymphocyte homeostasis, has been implicated in the development of lymphoid malignancies, exerts a tumour suppressor function, and contributes to drug‐induced cytotoxicity. We hypothesized that mutations of CD95 may occur in childhood B‐lineage acute lymphoblastic leukaemia (ALL), a disease known for its constitutive resistance towards CD95‐mediated apoptosis. We investigated 32 primary B‐lineage ALL of childhood and five B‐lineage ALL cell lines. All primary leukaemias expressed CD95 and bcl‐2 to a variable degree. Most of the leukaemias were resistant towards CD95‐mediated apoptosis. However, using SSCP analysis, no mutations in the coding and proximal promoter region could be detected. We conclude that the resistance towards CD95‐mediated apoptosis observed in most de novo B‐lineage ALL is not caused by mutations of the CD95 death receptor.
IgG from patients with antiphospholipid syndrome binds to platelets without induction of platelet activation
Authors: Ford  , Urbaniak  , Greaves
Published: March 2003

The clinical manifestations of the antiphospholipid syndrome (APLS) include arterial and venous thrombosis, thrombocytopenia and fetal loss, but the pathogenic mechanisms remain unclear. It has been hypothesized that platelet activation by autoantibody may be a pathogenic mechanism. We studied IgG binding, microparticle (mp) formation and P‐selectin expression by flow cytometry in normal platelets after incubation in serum from 11 patients with antiphospholipid antibodies and that from 10 normal healthy subjects. Levels of platelet‐associated IgG were significantly higher after incubation in patient sera (mean 17.2, range 2.0–75.0%) compared with normal sera (mean 2.0, range 1.2–3.7%, P < 0.05). Incubation of normal platelets in serum led to increased microparticle formation (P < 0.01) and P‐selectin expression (P < 0.05), compared with unstimulated platelets. There was no significant difference, however, between microparticle formation nor P‐Selectin expression induced by patient serum (mp 3.0 (1.6–5.0)%; P‐selectin 8.0 (4.0–16.6)%) versus normal serum (mp 3.2 (2.1–4.5)%; P‐selectin 10.1 (4.0–15.6); median (range)). Pre‐activation of platelets with subthreshold ADP concentrations or thrombin receptor activator peptide resulted in a small increase in microparticle formation, but there was still no significant difference between the effects of patient and control sera. Despite the presence of platelet membrane binding IgG in serum from 5/11 patients with antiphospholipid antibodies, there was no evidence for associated enhanced platelet‐activating ability. This study supports antiplatelet reactivity in antiphospholipid syndrome, but not a direct platelet‐activating role for platelet‐directed autoantibodies.
Thiamine‐responsive myelodysplasia
Authors: Bazarbachi Ali , Muakkit Samar , Ayas Mouhab , Taher Ali , Salem Ziad , Solh Hassan , Haidar Joud H.
Published: March 2003

The triad of thiamine‐responsive anaemia, diabetes mellitus and deafness has been reported in 15 patients with macrocytic anaemia, sometimes associated with moderate thrombocytopenia. The bone marrow aspirate usually shows megaloblastic changes and ringed sideroblasts. However, tri‐lineage myelodysplasia has never been reported. We describe two patients who presented with diabetes, deafness and thiamine‐responsive pancytopenia. Bone marrow aspirate and biopsy were typical of tri‐lineage myelodysplasia. These findings suggest that thiamine may have a role in the regulation of haemopoiesis at the stem cell level. We propose the term ‘thiamine‐responsive myelodysplasia’ rather than that of thiamine‐responsive anaemia.
Dendritic cells derived from bone marrow and CD34+ selected blood progenitor cells of myeloma patients, cultured in serum‐free media, do not contain the Kaposi sarcoma herpesvirus genome
Authors: Mitterer  , Mair  , Gatti  , Sheldon  , Vachula  , Coser  , Schultz
Published: March 2003

The aim of our study was to test if dendritic cells contain the KSHV genome. CD34+ peripheral blood progenitor cells (PBPC) and bone marrow mononuclear cells were cultured in X‐VIVO 15 medium supplemented with GM‐CSF and TNF‐alpha in gas‐permeable containers. Dendritic cells were identified morphologically and immunophenotypically. The KSHV genome was not identified in any of the cases using a nested primer PCR approach. Serological analysis corroborated the molecular findings: no antibodies for KSHV were found in any of the multiple myeloma patients. These data are of importance when considering use of DC for therapeutic approaches in multiple myeloma.
Autologous transplantation in chronic myeloid leukaemia using peripheral blood stem cells
Authors: Singer  , Franklin  , Clark  , Chalmers  , Kelsey  , Newland  , Sproul  , Crotty  , Mccann  , Goldstone  , Mcbride  , Hepplestone  , Watson  , Sharp  , Tansey
Published: March 2003

Forty‐three patients with chronic myeloid leukaemia in first chronic phase were recruited to study intensive chemotherapy (idarubicin plus cytarabine; IdAC) followed by collection of peripheral blood stem cells (PBSC) in the recovery phase. PBSC autografting was performed on 32 patients. One patient died during mobilization and three died following autograft. All procedural deaths occurred in patients who received IdAc more than a year from diagnosis. Nine further patients died, eight following progression of CML. 72% of transplanted patients showed a major cytogenetic response but most cases have returned to Philadelphia‐positive haemopoiesis. 62% of autografted patients remain alive (median survival from diagnosis 52 months). Four of the 11 patients who did not receive a transplant remain in chronic phase.
Mutational screening of thrombopoietin receptor gene (c‐mpl) in patients with congenital thrombocytopenia and absent radii (TAR)
Authors: Strippoli Pierluigi , Savoia Anna , Iolascon Achille , Tonelli Roberto , Savino Maria , Giordano Paola , D'avanzo Michele , Massolo Fausta , Locatelli Franco , Borgna Caterina , DE Mattia Domenico , Zelante Leopoldo , Paolucci Guido , Bagnara Gian Paolo
Published: March 2003

Thrombocytopenia with absent radii (TAR) is a rare autosomal recessive disease characterized by hypomegakaryocytic thrombocytopenia and bilateral radial aplasia. We performed mutational screening of coding and promoter regions of the c‐mpl gene, encoding thrombopoietin (TPO) receptor, by sequence analysis in four unrelated patients affected by TAR syndrome. Our results indicate that c‐mpl gene mutations are not a common cause of thrombocytopenia in TAR syndrome.
Kupffer cell staining by an HFE‐specific monoclonal antibody: implications for hereditary haemochromatosis
Authors: Bastin Judy M , Jones Margaret , O'callaghan Christopher A. , Schimanski Lisa , Mason David Y. , Townsend Alain R. M.
Published: March 2003

Hereditary haemochromatosis is an inherited disorder of iron absorption that leads to excessive iron storage in the liver and other organs. A candidate disease gene HFE has been identified that encodes a novel MHC class I like protein. We report the development of a monoclonal antibody (HFE‐JB1) specific for recombinant refolded HFE protein. The antibody immunoprecipitates a 49 kD protein from the cell line U937, a histiocytic lymphoma. It binds HFE but does not recognize other recombinant non‐classic MHC class I proteins (HLA‐E, F and G), nor does it react with a variety of recombinant classic class I MHC molecules. COS cells transfected with HFE in culture are stained specifically. The immunohistochemical staining pattern in human tissues is unique and can be defined as a subset of the transferrin receptor positive cells. In the liver HFE protein was shown to be present on Kupffer cells and endothelium (sinusoidal lining cells), but absent from the parenchyma. Kupffer cells from an untreated C282Y HH patient failed to stain with the antibody. In the normal gut scattered cells in the crypts are stained. HFE was also present on capillary endothelium in the brain (a site of high levels of transferrin receptor) and on scattered cells in the cerebellum and cortex. These results raise interesting questions concerning the function of HFE in the control of body iron content and distribution.
Elevated serum levels of soluble vascular cell adhesion molecule‐1 (sVCAM‐1) closely reflect tumour burden in chronic B‐lymphocytic leukaemia
Authors: Christiansen Ilse , Sundström Christer , Tötterman Thomas H.
Published: March 2003

The present study is the first to report elevated serum levels of soluble (s)VCAM‐1 in B‐cell chronic lymphocytic leukaemia (B‐CLL). A large cohort of 106 untreated patients was studied. sVCAM‐1 was compared to known prognostic serum markers (soluble (s)ICAM‐1; lactate dehydrogenase, LDH; sCD23; thymidine kinase, TK; β2microglobulin, β2m). The serum levels of sVCAM‐1 reflected tumour burden as expressed by Binet/Rai stages more closely than any other marker. sVCAM‐1 also reflected the kinetics of the disease as revealed by lymphocyte doubling time. sVCAM‐1 was the only one of the studied markers which showed elevated levels in smouldering disease compared to controls. sVCAM‐1, sICAM‐1 and sCD23 (but not LDH, TK, β2m) separated smouldering from non‐smouldering B‐CLL. Only sICAM‐1, sCD23 and TK added independent prognostic information for survival to that of stage and lymphocyte doubling time.

The expression of both adhesion molecules was examined in lymph node and splenic specimens. VCAM‐1 and ICAM‐1 were overexpressed by vascular endothelium and stroma, but the intensity of expression correlated poorly with serum levels of the soluble molecules.

In conclusion, serum levels of sVCAM‐1 correlated with tumour burden and other prognostic markers in B‐CLL. VCAM‐1 was overexpressed in tumour tissue as was ICAM‐1. sVCAM‐1 could prove a valuable marker in younger early‐stage patients eligible for therapeutic trials.
Content of long‐term culture‐initiating cells, clonogenic progenitors and CD34+cells in apheresis harvests of normal donors for allogeneic transplantation, and in patients with acute myeloid leukaemia or multiple myeloma
Authors: Kasper C. , Ryder W. D. J. , Dürig J. , Nagesh K. , Scarffe J. H. , Beelen D. W. , Schaefer U. W. , Chang J. , Testa N. G.
Published: March 2003

Using a limiting dilution assay the frequency of long‐term culture‐initiating cells (LTC‐IC) in the apheresis products following mobilization by granulocyte‐colony stimulating factor (G‐CSF) with or without chemotherapy from 14 normal donors (ND) for allogeneic bone marrow transplantation, 16 patients with multiple myeloma (MM) and 15 patients with acute myeloid leukaemia (AML), where the aphereses were intended for autologous transplantation, were compared.

The estimated median incidences of LTC‐IC in the first apheresis products from ND, MM and AML were 1/3289, 1/1775 and 1/13 075 mononuclear cells (MNC) respectively. The patients with AML had a significantly lower incidence compared with the other two groups (P < 0.0001). There was a positive correlation between the incidence of LTC‐IC and the number of CD34+cells, the number of GM‐CFC, and the number of BFU‐E. The positive association with GM‐CFC or BFU‐E was weaker. In these experiments the percentage of CD34+cells was the best predictor for the frequency of LTC‐IC in the peripheral blood progenitor cells (PBPC). In eight cases of MM the LTC‐IC assay was performed for both the first and second harvest. All cases had a lower LTC‐IC frequency in the second harvest compared with the first, an average of 23% (13–42%, 95% confidence interval) and this reduction was statistically significant (P < 0.001); CD34+cells were also lower (P < 0.001).
Determination of the breakpoint of the common α‐thalassaemia deletion in Filipinos in Hawaii
Authors: Hunt John A. , Lee Lara , Donlon Timothy A. , Hsia Y. Edward
Published: March 2003

The region of the crossover causing the Filipino type of α thalassaemia has been determined by examining similarity between the regions which had been indicated as involved in the crossover points by restriction mapping, using the published α‐globin region DNA sequence. The crossover point was found in 21 base pairs between two Alu sequences using PCR primers flanking these Alu sequences. A simple PCR multiplex assay has been devised to detect heterozygotes and homozygotes.
Bone marrow transplants for paroxysmal nocturnal haemoglobinuria
Authors: Sašo Radovan , Marsh Judith , Čevreska Lidija , Szer Jeffrey , Gale Robert Peter , Rowlings Philip , Passweg Jakob , Nugent Melodee , Luzzatto Lucio , Horowitz Mary , Gordon‐Smith Edward
Published: March 2003

Paroxysmal nocturnal haemoglobinuria (PNH) is a rare clonal haematological disorder characterized by intravascular haemolysis and increased risk of thrombosis. PNH is associated with bone marrow failure syndromes including aplastic anaemia, myelodysplasia and leukaemia. Bone marrow transplants are sometimes used to treat PNH, but small series and reporting biases make assessment of transplant outcome difficult. The outcome of 57 consecutive allogeneic bone marrow transplants for PNH reported to the International Bone Marrow Transplant Registry (IBMTR) between 1978 and 1995 was analysed. The 2‐year probability of survival in 48 recipients of HLA‐identical sibling transplants was 56% (95% confidence interval 49–63%). Two recipients of identical twin transplants remain alive 8 and 12 years after treatment. One of seven recipients of alternative donor allogeneic transplants is alive 5 years after transplant. The most common causes of treatment failure were graft failure and infections. Our results indicate that bone marrow transplantion can restore normal bone marrow function in about 50% of PNH patients.
Consistent detection of CALM‐AF10 chimaeric transcripts in haematological malignancies with t(10;11)(p13;q14) and identification of novel transcripts
Authors: Narita Masami , Shimizu Kimiko , Hayashi Yasuhide , Taki Tomohiko , Taniwaki Masafumi , Hosoda Fumie , Kobayashi Hirofumi , Nakamura Hideo , Sadamori Naoki , Ohnishi Hiroaki , Bessho Fumio , Yanagisawa Masayoshi , Ohki Misao
Published: March 2003

The t(10;11)(p13‐14;q14‐21) is a rare but recurring translocation associated with acute lymphoblastic leukaemia (ALL) and acute myeloid leukaemia (AML). Recently the CALM gene was cloned from the t(10;11) breakpoint of U937 and fused to AF10, a putative transcription factor, which had been identified as one of the fusion partners of the MLL gene. In order to define the involvement of these genes in primary leukaemias and cell lines with t(10;11), we analysed the expression of fusion transcripts by reverse transcriptase‐polymerase chain reaction (RT‐PCR) in five patient samples including ALL, AML and lymphoblastic lymphoma, and three monocytic cell lines (P31/Fujioka, KP‐Mo‐TS and U937). The CALM‐AF10 fusion transcript was detected in all samples; however, the AF10‐CALM fusion was not detected in two patient samples and one cell line. In RT‐PCR analysis there were six isoforms of the CALM‐AF10 fusion transcripts and five of AF10‐CALM fusion transcripts. We also detected novel transcripts in U937. Sequence analysis revealed that all these isoforms had in‐frame junctions and that some of them resulted from alternative splicing at different exons of CALM and others from different breakpoints at CALM and/or AF10. There were at least two different breakpoints of CALM and three of AF10 gene. Our results suggest that the CALM‐AF10 fusion gene is a constant feature and is involved in the pathogenesis of haematological malignancies with t(10;11)(p13‐14;q14‐21), showing various and often multilineage phenotypes. Thus, t(10;11) needs to be investigated by RT‐PCR for identification of the genes involved.
Myeloid antigen co‐expression in childhood acute lymphoblastic leukaemia: relationship with in vitro drug resistance
Authors: Den Boer M. L. , Kapaun P. , Pieters R. , Kazemier K. M. , Janka‐Schaub G. E. , Veerman A. J. P.
Published: March 2003

Contradictory data have been reported about the prognostic value of myeloid antigen co‐expression (My+) in childhood acute lymphoblastic leukaemia (ALL). In the present study the methyl thiazol tetrazoliumbromide (MTT) assay was used to compare the in vitro cytotoxicity of 14 drugs between 60 My+ (CD13+ and/or CD33+) and 107 My− ALL children at initial diagnosis. P‐glycoprotein (P‐gp), multidrug resistance‐associated protein (MRP), major vault protein/lung resistance protein (LRP) and the intracellular daunorubicin concentration were studied by flow cytometry. My+ ALL samples were significantly more resistant, i.e. between 1.1‐ and 2.9‐fold, to daunorubicin, doxorubicin, idarubicin, mitoxantrone, vincristine, 6‐thioguanine, 6‐mercaptopurine, teniposide, etoposide and ifosfamide compared with My− ALL samples. My− and My+ ALL did not significantly differ in sensitivity to prednisolone, dexamethasone, l‐asparaginase and cytarabine. Comparable results were found when only common and preB ALL cases were analysed. Drug resistance in My+ ALL was not related to increased expression of P‐gp, MRP or LRP compared with My− ALL (ratio My+/My−:P‐gp 0.8, MRP 1.0, LRP 1.1). Accumulation and retention of daunorubicin did not significantly differ between My− and My+ ALL cells (ratio My+/My−: accumulation 1.2, retention 1.3). Therefore the nature of drug resistance in My+ ALL remains unknown. The lack of prognostic value for My+ in childhood ALL may be explained by the responsiveness of My+ ALL to glucocorticoids, l‐asparaginase and cytarabine. In addition, the currently intensive treatment regimens may apply drug doses which are simply high enough to overcome the mild resistance to anthracyclines, mitoxantrone, vincristine, thio‐purines, epipodophyllotoxins and ifosfamide in childhood My+ ALL.
Secondary T‐acute lymphoblastic leukaemia mimicking blast crisis in chronic myeloid leukaemia
Authors: Dawson  , Slater  , Hagemeijer  , Langerak  , Willemze  , Kluin‐Nelemans
Published: March 2003

A 34‐year‐old man with chronic myeloid leukaemia (CML) firstly developed a lymphoid blast crisis of B‐cell type. After a second chronic phase which lasted for > 4 years with maintenance chemotherapy of hydroxyurea, 6‐mercaptopurine and methotrexate, he developed a T‐cell acute lymphoblastic leukaemia of TcR‐γδ+ type. Cytogenetic analysis revealed disappearance of the t(9;22) translocation and appearance of new abnormalities consistent with the diagnosis secondary acute leukaemia. To our knowledge, secondary leukaemia in CML has not previously been reported.
Impact of obstetric factors on cord blood donation for transplantation
Authors: Donaldson Craig , Armitage W. John , Laundy Val , Barron Clive , Buchanan Ruth , Webster Jill , Bradley Ben , Hows Jill
Published: March 2003

Recent reports have shown that low nucleated cell dose significantly decreases survival after cord blood transplantation. Prior to starting clinical cord blood banking we investigated the impact of obstetric factors on cell dose and volume of cord blood donations. Cord blood was obtained from 114 normal full‐term deliveries. Mean volume collected was 93.5 ml, mean total nucleated cell count (TNC) was 13.1 × 108. Statistical analysis was by backwards stepwise regression. Significant factors affecting nucleated cell yield were volume of blood collected (P < 0.001), length of gestation (P < 0.0001), time from delivery of the infant to cord clamping (P = 0.018) and total length of labour (P = 0.002). In clinical cord blood banking we have successfully used these findings for pre‐collection assessment of placentae. Out of 476 cord blood donations subsequently collected for banking, only 29 (6.1%) have been discarded due to low volume. The mean TNC of the 409 banked units following volume reduction was 10.1 × 108. Despite careful optimization of collection, processing and storage techniques, cell dose still limits cord blood transplantation to smaller recipients.
Identification of X‐linked severe combined immunodeficiency by mutation analysis of blood and hair roots
Authors: Ting Suk See , Leigh Don , Lindeman Robert , Ziegler John B.
Published: March 2003

Severe combined immunodeficiency is a heterogenous syndrome of varied genetic origins of which the X‐linked type is the commonest (XSCID). The most sensitive method for diagnosis of XSCID in the absence of X‐linked inheritance pattern is by mutation analysis.

In this report we have performed mutation analysis in 13 unrelated boys transplanted (BMT) for SCID without a known cause to determine the frequency of XSCID. Five boys had an affected male relative. We also assessed the utility of hair roots for children without pre‐transplant blood stored for mutation analysis since donor genotype was expressed in peripheral blood post BMT. Screening was performed by analysis of single‐strand conformational polymorphism (SSCP) followed by sequencing of candidate exons.

Mutations were found in 11 cases, of which six were sporadic, and maternal mosaicism was found in one family. Three mothers of the six sporadic cases were identified as carriers. The majority (6/8) of boys with SCID had γc deficiency despite the absence of X‐linked inheritance pattern. The significant frequency of de novo mutations and the occurrence of maternal germline mosaicism highlights the importance of mutation analysis. The strategy of using DNA from hair roots was particularly valuable where no pre‐transplant blood was stored. Characterization of the mutations will also enable research into the correction of these genetic defects.
Circadian variation of granulocyte colony stimulating factor levels in man
Authors: Jilma B. , Hergovich N. , Stohlawetz P. , Eichler H.‐G. , Bauer P. , Wagner O. F.
Published: March 2003

Glucocorticoids dose‐dependently increase plasma levels of granulocyte colony stimulating factor (G‐CSF). Based on the marked circadian rhythm of cortisol levels, we hypothesized that plasma levels of G‐CSF may also show a diurnal rhythm. A prospective study was conducted in 12 healthy young volunteers. Blood samples were obtained every 2 h over 24 h. G‐CSF levels averaged 18.0 ng/l (CI 13.1–22.9) at 8.00 am, increased continuously and reached peak values at 10.00 p.m. Individual harmonic regression analysis showed a clear circadian rhythm. The individual differences between nadir and peak levels averaged 54% (CI 43–65%). This pronounced diurnal rhythm of G‐CSF levels may help understand the circadian changes in circulating stem cells, bone marrow DNA synthesis, or bone marrow toxicity induced by some chemotherapeutic agents.
The CD69 early activation molecule is overexpressed in human bone marrow mast cells from adults with indolent systemic mast cell disease
Authors: Díaz‐Agustín Beatriz , Escribano Luis , Bravo Pilar , Herrero Sonia , Nuñez Rosa , Navalón Raquel , Navarro Lourdes , Torrelo Antonio , Cantalapiedra Alberto , Del Castillo Luz , Villarrubia Jesús , Navarro José L. , San Miguel Jesús F. , Orfao Alberto
Published: March 2003

We have analysed the quantitative expression of surface CD69 antigen on human mast cells (MC), from both normal and pathological bone marrow (BM) samples, using flow cytometry. Our major aim was to analyse whether CD69 is constitutively expressed by normal BMMC and to explore the possible differences between CD69 expression by BMMC from normal controls and patients suffering from different pathological conditions.

The constitutive expression of surface CD69 was clearly demonstrated in BMMC; however, systemic mast cell disease (SMCD) patients showed significantly higher levels of surface CD69 expression than healthy controls (P < 0.001), chronic lymphocytic leukaemia (P = 0.001), monoclonal gammopathy of unknown significance (P < 0.001), multiple myeloma (P < 0.001) patients, and myelodysplastic syndromes (P = 0.002). Furthermore, almost no overlap between the levels of CD69 expression on BMMC was observed between SMCD cases and the remaining groups of individuals except for the paediatric mastocytosis group (P > 0.05). From the other groups of patients, monoclonal gammopathy of unknown significance (P = 0.04), myelodysplastic syndromes (P = 0.03) and paediatric mastocytosis (P = 0.003) cases showed a significantly higher expression of surface CD69 as compared to normal subjects.

In summary, our findings show that the CD69 antigen is overexpressed in SMCD patients.
Hepatosplenic candidiasis in patients with acute leukaemia
Authors: Sallah Sabah , Semelka Richard C. , Wehbie Robert , Sallah Waddah , Nguyen Nam P. , Vos Paul
Published: March 2003

A retrospective study of 23 patients with acute leukaemia and hepatosplenic candidiasis (HSC) was conducted to evaluate clinical treatment characteristics in terms of amount and duration of antifungal agents and to assess treatment outcome.

Patients were admitted to two major tertiary care centres between 1990 and 1998. The diagnosis of HSC was based on clinical, blood cultures, histologic and imaging studies. Patients were treated with amphotericin B without interruption of the planned chemotherapy regimens. Serial magnetic resonance imaging (MRI) studies were the main tool for following patients' response and activity of the fungal lesions in conjunction with clinical and laboratory parameters.

Treatment with amphotericin B was continued until resolution of all clinical symptoms and signs attributable to HSC, obtaining negative blood cultures and the appearance of at least healed lesions on MRI. Amphotericin B was discontinued in four patients because of severe nephrotoxicity (two patients), or continuous fever and persistent fungal lesions on MRI (two patients). Amphotericin B lipid complex (ABELCET) was successfully used as salvage therapy for these refractory patients. Four patients died with evidence of HSC despite treatment and supportive measures. The response rate for treatment of HSC was 82%. The mean total dose of amphotericin B including empirical treatment was 4 g and the median duration of treatment for responding patients was 112 d. The median number of days of anti‐ fungal treatment before the disappearance of fever was 19 d.

Our results confirmed the need for protracted courses of antifungal agents for the successful eradication of HSC. Chemotherapy for the underlying disorder should not be interrupted or delayed in order to treat HSC.
Fas ligand‐induced apoptosis of hepatocytes in natural killer cell leukaemia
Authors: Tani Ryuzo , Ozaki Shuji , Kosaka Masaaki , Fujiwara Soichiro , Shibata Hironobu , Wakatsuki Shingo , Matsumoto Toshio
Published: March 2003

Neoplastic natural killer (NK) cells overexpress Fas ligand (FasL), which may cause damage of Fas‐bearing tissues. We report a patient with NK cell leukaemia who developed liver injury after pharyngitis. The NK leukaemic cells expressed functional FasL. In addition to soluble FasL, serum levels of interleukin‐6 and interferon‐γ were increased dramatically when liver injury was aggravated. Moreover, hepatocytes expressed Fas and apoptotic hepatocytes were detected in the portal areas. These findings are consistent with the notion that inflammatory cytokines enhance the sensitivity to FasL and trigger apoptosis of hepatocytes in NK cell malignancies.
Results at a single centre of immunosuppression with cyclosporine A in 66 children with aplastic anaemia
Authors: Maschan Alexei , Bogatcheva Natalia , Kryjanovskii Oleg , Shneider Maria , Litvinov Dimitry , Mitiushkina Tatiana , Timakov Andrei , Timakova Maria , Samotchatova Elena , Rumiantsev Alexander
Published: March 2003

A retrospective analysis of cyclosporine A (CsA) monotherapy administered to 66 children with aplastic anaemia (AA) at a single centre was carried out. The study was conducted on 66 children (F34/M32) with a median age of 10.5 years. 30 children (45%) achieved complete or partial remission within a median of 8 weeks. The response rate was 3/19 (16%) in cases of very severe aplastic anaemia (vSAA), 16/34 (47%) in severe aplastic anaemia (SAA) and 11/13 (85%) in nonSAA. 10 remitters (33%) relapsed after CsA cessation or dose tapering and six of those responded again on CsA alone. The only variable predictive for response was granulocyte count before treatment. The actuarial probability of survival was 51% at 4 years (85% in moderate AA, 50% in SAA and 32% in vSAA). The optimal dosage of CsA has yet to be established.
Efficacy and toxicity of IFN‐α2b combined with cytarabine in chronic myelogenous leukaemia
Authors: Lindauer  , Domkin  , Döhner  , Kolb  , Neubauer  , Huhn  , Kreiter  , Koch  , Huber  , Aulitzky  , Fischer
Published: March 2003

Newly diagnosed chronic myelogenous leukaemia (CML) patients (n = 65) were treated with interferon (IFN)‐α2b (5 × 106 IU/d s.c.) combined with monthly courses of cytarabine (20 mg/d s.c. for 14 d). Median age of patients enrolled was 45 years. The endpoints of the study were clinical efficacy and toxicity. The survival rates at 3 years and 5 years were 77% and 56%, respectively. The rate of complete haematological response was 60%. Evaluation of cytogenetic response was available in 29/65 patients. A complete cytogenetic response was seen in 3/29 patients (10%). W.H.O. toxicity grade 3–4 occurred in only 22/523 evaluable treatment cycles. Since the study protocol required intermittent or definitive discontinuation of cytarabine in case of moderate leucopenia (white blood cells (WBC) <5 × 109/l), combined cytopenia (WBC < 5 × 109/l, platelets <100 × 109/l), and isolated moderate thrombocytopenia (<100 × 109/l), the drug had to be discontinued temporarily or definitively in 200 cycles and the dose of cytarabine had to be reduced in 35 cycles. Thus, only 25% of the planned dose of cytarabine could be administered. At this dosage it would appear that cytarabine had no effect on survival and did not improve remission rates.

We conclude that a clinical benefit for the addition of cytarabine to the treatment of CML with IFN might only be achieved by the administration of a higher cumulative dose of cytarabine, suggesting that lower leucocyte counts of 2–4 × 109/l have to be tolerated.
Pregnancy in bone marrow failure syndromes: Diamond‐Blackfan anaemia and Shwachman‐Diamond syndrome
Authors: Alter Blanche P. , Kumar Manjusha , Lockhart Lillian L. , Sprinz Philippa G. , Rowe Thomas F.
Published: March 2003

Pregnancy in bone marrow failure syndromes has risk to mother and fetus. There are fewer than 30 reports of cases with Diamond‐Blackfan anaemia (DBA), and none with Shwachman‐Diamond syndrome (SD). We report two DBA and one SD cases. One DBA mother received transfusions intra‐partum, and the other only post‐partum. Both required caesarean sections (C‐sections) for failure of labour to progress and severe pre‐eclampsia respectively. Both subsequently resumed pre‐pregnancy steroid‐induced control of anaemia. ~40% of DBA pregnancies required maternal transfusions; 25% delivered by C‐section. The SD patient also had Ehlers‐Danlos (ED) syndrome and urticaria pigmentosa (UP). Her blood counts were adequate until week 38, when the platelet count dropped and a C‐section was performed. Pregnancy management in marrow failure disorders requires obstetricians with expertise in high‐risk pregnancies, and haematologists with experience with marrow failure syndromes.
Haemopoietic progenitor cell lines generated by the myeloproliferative leukaemia virus: a model system to analyse murine and human lineage‐affiliated genes
Authors: Thomson Andrew M. , Roberts Nigel A. , Sloane Stanley Jacqueline A. , Souyri Michèle , Wood William G.
Published: March 2003

Multipotential progenitor and stem cells occur with a low frequency in haemopoietic tissue. As a result, it is often difficult to obtain sufficient numbers of cells to undertake many of the assays that would be informative about the molecular events involved in the regulation of lineage‐affiliated genes within these multipotent cells. To circumvent this problem, we have used the myeloproliferative leukaemia virus (MPLV) to generate a phenotypically diverse array of haemopoietic progenitors from adult mouse bone marrow and embryonic blood. These cells could be expanded to perform a variety of analyses that would not previously have been possible using analogous primary cells. The validity of these assays was supported by the observation that the phenotype of several MPLV‐infected lines was very similar to previously described primary haemopoietic progenitor cells. By using mice transgenic for the human alpha and beta globin gene clusters, we have shown that human genes may also be investigated. In addition, this strategy has a wide potential applicability including the rescue of haemopoietic progenitors from mouse embryos lacking genes critical for their survival as well as the study of any haemopoietic gene for which an appropriate transgenic mouse is available.
Cys97→Tyr mutation in the glycoprotein IX gene associated with Bernard‐Soulier syndrome
Authors: Kunishima Shinji , Tomiyama Yoshiaki , Honda Shigenori , Kurata Yoshiyuki , Kamiya Tadashi , Ozawa Kazuo , Saito Hidehiko
Published: March 2003

Bernard‐Soulier syndrome (BSS) is an autosomal recessive bleeding disorder due to quantitative or qualitative abnormalities in the glycoprotein (GP) Ib/IX/V complex, the platelet receptor for von Willebrand factor. We describe here the genetic basis of the disorder in a patient with BSS. Flow cytometric analysis of the patient's platelets showed a greatly reduced GPIbα and completely absent GPIX surface expression. Immunoblot analysis disclosed greatly reduced GPIbα and residual amounts of GPIbβ and GPIX in the platelets. DNA sequencing analysis revealed the patient to be homozygous for a novel missense mutation in the GPIX gene that converts Cys (TGT) to Tyr (TAT) at residue 97. Transient transfection studies confirmed that mutant GPIX was not expressed on the transfected cells, showing that the mutation was responsible for the BSS phenotype observed in the patient.
Low frequency of the TEL/AML1 fusion gene in acute lymphoblastic leukaemia in Spain
Authors: García‐Sanz  , Alaejos  , Orfão  , Rasillo  , Chillón  , Tabernero  , Mateos  , López‐Pérez  , González  , Balanzategui  , González  , San Miguel
Published: March 2003

We report on a series of Spanish patients with acute lymphoblastic leukaemia in whom the t(12;21) [TEL/AML1] translocation could not be identified with two sensitive techniques: reverse transcript‐polymerase chain reaction (RT‐PCR) and fluorescence in‐situ hybridization (FISH). 101 cases were analysed: 38 children (29 B‐cell precursor; nine T‐cell precursor) and 63 adults (48 B‐cell precursor; 15 T‐cell precursor). Specific RT‐PCR to amplify the TEL/AML1 fusion transcript was negative in all 101 cases. Moreover, all 38 paediatric samples were also negative by interphase FISH analysis for the presence of the TEL/AML1 fusion. These results suggest the existence of geographic/race variations in the genotype of acute lymphoblastic leukaemia (ALL).