bluuebunny/crossref_metadata_2025 huggingface dataset:
Levels of complement regulatory proteins, CD35 (CR1), CD46 (MCP) and CD55 (DAF) in human haematological malignancies
Authors: Hara Tomoko , Kojima Ayako , Fukuda Hiroko , Masaoka Toru , Fukumori Yasuo , Matsumoto Misako , Seya Tsukasa
Published: March 2008

Summary. Levels of the membrane complement regulatory proteins. C3b/C4b receptor (CR1, CD35), membrane cofactor protein (MCP, CD46), and decay‐accelerating factor (DAF, CD55), expressed on cells from patients with haematological malignancies and normal subjects were assessed by flowcytometry using the respective monoclonal antibodies (mAbs). All myeloid and most lymphoid leukaemia samples tested were CR1‐negative: two of the 42 leukaemia samples expressed minute amounts of CR1. Lack of CR1 in leukaemia cells was confirmed with two mAbs raised against CR1, 31R, and 243R, which recognized different epitopes and induced different degrees of CR1‐mediated fluorescent shift on flow‐cytometry in granulocytes and erythrocytes. MCP was increased in most chronic myelogenous leukaemia (CML) and chronic lymphocytic leukaemia (CLL), and was also increased in majority of acute nonlymphocytic leukaemia (ANLL), acute lymphocytic leukaemia (ALL) and non‐Hodgkin's lymphoma (NHL). Levels of DAF were also high in CML and CLL, and were variable in other types of leukaemia: some were DAF‐negative while others expressed extremely high levels of DAF. In CML patients, the high level of MCP and the lack of CR1 were normalized after medical treatment. These results are in agreement with the data obtained with human leukaemia cell lines, and support the hypothesis that CR1 is essentially a differentiated cell antigen and that a high level of MCP reflects some malignant transformation or an immature stage in blood cells.
Cellular and humoral immune responses in haemophiliacs after vaccination against tick‐borne encephalitis
Authors: Wolf Hermann M. , Pum Martina , Jáger Rita , István Lajos , Mannhalter Josef W. , Eibl Martha M.
Published: March 2008

Summary. The primary immune response to a viral antigen (tick‐borne encephalitis, TBE) has been determined in haemophiliacs. Twelve HIV‐negative and four clinically asymptomatic, HIV‐positive haemophiliacs as well as 16 age‐matched healthy controls were included in the study. Antibody responses after TBE vaccination were comparable in HIV‐negative haemophiliacs and controls: however, antibody titres in HIV‐infected haemophiliacs were significantly lower after completion of the three‐dose vaccination schedule (geometric mean reciprocal antibody titres (SEM): controls. 193 (1.37), HIV‐positive haemophiliacs. 13 (2.18), P< 0.005). TBE vaccination failed to induce a T cell proliferative response in the HIV‐positive haemophiliacs. While in HIV‐negative patients the antigen‐specific lymphoproliferative responses after primary and one booster vaccination were comparable to those of the controls, cellular responses were decreased in HIV‐negative haemophiliacs following a second booster immunization 19 months after primary immunization (3H‐thymidine incorporation, delta dpm, mean ± SEM: controls, 34662 ± 7129, HIV‐negative haemophiliacs, 14339±7420, P<0.005). As the protective mechanisms for TBE infection are not yet completely understood, further work will be necessary to determine whether the decreased capacity to mount a sufficient long‐term cellular memory response in HIV‐negative haemophiliacs might be important for the protective effect of TBE vaccination in this population.
Inhibition of thrombin generation by heparin and low molecular weight (LMW) heparins in the absence and presence of platelet factor 4 (PF4)
Authors: Padilla Ana , Gray Elaine , Pepper Duncan S. , Barrowcliffe Trevor W.
Published: March 2008

Summary. The ability of several low molecular weight (LMW) heparins and unfractionated heparin (UFH) to inhibit thrombin generation, and their anti‐Xa and anti‐IIa activities, were measured in the absence and presence of platelet factor 4 (PF4).

The LMW heparins studied were 2–5 times less potent, on a weight basis, than UFH as inhibitors of thrombin generation in platelet‐poor plasma: the inhibition of thrombin generation by LMW heparins correlated better with their anti‐IIa activity (r= 0–98) than with their anti‐Xa activity (r= 0–69). At low concentrations of PF4. the activity of LMW heparins in the thrombin generation test was neutralized less than that of UFH. but at higher PF4 concentrations all their activities could be neutralized except in anti‐Xa assays.

These observations support the hypothesis that anti‐IIa activity is important for inhibition of thrombin generation by LMW heparins in vitro. However, when all the anti‐IIa activity of LMW heparins was neutralized by PF4. considerable inhibitory activity remained in thrombin generation and anti‐Xa assays, indicating that a portion of the anti‐Xa activity of LMW heparins also contributes towards inhibition of thrombin generation.
Effects of bryostatin 1 and rGM‐CSF on the metabolism of 1‐β‐d‐arabinofuranosylcytosine in human leukaemic myeloblasts
Authors: Grant Steven , Jarvis W. David , Turner Amy J. , Wallace Hugh J. , Pettit George R.
Published: March 2008

Summary. The effects of the protein kinase C activator bryostatin 1, either with or without recombinant granulocyte‐macrophage colony stimulating factor (rGM‐CSF) were examined with respect to the in vitro metabolism of ara‐C in leukaemic myeloblasts obtained from 10 patients with acute myelogenous leukaemia (AML). Coincubation of cells with 12·5 × 10−9m bryostatin 1 and 10−5m ara‐C for 4 h resulted in a significant increase in ara‐CTP formation (compared to controls) in 6/10 specimens (mean increase 106%: range 38–255%), and no change in the remainder. In contrast, coincubation of cells with 1·25 ng/ml rGM‐CSF resulted in a significant increase in only one specimen, and decreases in two. Bryostatin 1 also significantly increased ara‐C DNA incorporation in 6/9 evaluable samples, including two which did not display an increase in ara‐CTP formation. Coincubation of cells with both bryostatin 1 and rGM‐CSF did not lead to a further increase in ara‐CTP formation or ara‐C DNA incorporation compared to values obtained with either agent alone. Finally, exposure of blasts to bryostatin 1 for 24 h before ara‐C led to an increase in ara‐CTP formation in 3/8 additional specimens, and a decrease in one sample displaying evidence of bryostatin 1 ‐induced macrophage differentiation. Incubation of cells with both rGM‐CSF and bryostatin 1 for this period resulted in ara‐CTP levels equivalent to those obtained with bryostatin 1 alone. These studies indicate that while bryostatin 1 exerts a heterogeneous effect on ara‐C metabolism in leukaemic myeloblasts, it is capable of potentiating ara‐C phosphorylation in a subset of patient samples, including some that do not exhibit an increase in response to rGM‐CSF. They also raise the possibility that bryostatin 1‐induced potentiation of ara‐C metabolism in some leukaemic cells may contribute, at least in part, to the antileukaemic efficacy of this drug combination.
Mechanisms of inhibition of T cell IL‐2 secretion by factor VIII concentrates
Authors: Wadhwa Meenu , Dilger Paula , Tubbs Jill , Barrowcliffe Trevor , Mahon Bernard , Thorpe Robin
Published: March 2008

Summary. We have continued our previous study of the inhibitory effects of factor VIII concentrates on IL‐2 secretion by T cells. Experiments with an extended range of products confirm our previous conclusion that some but not all low, intermediate and high purity concentrates possess inhibitory activity on IL‐2 secretion. The inhibition occurs almost immediately after addition of factor VIII concentrate and it was not possible to adsorb inhibitory activity with activated or non‐activated cells; this suggests that the mechanism of inhibition involves interference with early T cell activation events rather than simple blocking of cell surface components by inhibitory molecules.

The inhibitory components were shown to reside in different molecular weight fractions of concentrates. A strongly inhibitory component of approximately 200 kD and a minor species of approximately 60 kD were identified in strongly inhibitory concentrates. Some products contained a dialysable inhibitory substance which is most likely a salt as it was also present in some formulation buffers. The proportions of the inhibitory components varied widely between products.

We have found that the pattern of inhibition using in vitro systems reflects that observed using a mouse in vivo antigen challenge method. In addition we have shown that the previously reported concentrate mediated inhibition of lectin induced low affinity IL‐2 receptor (CD25) is mainly a consequence of diminished IL‐2 secretion rather than a ‘direct’ effect on CD2 5 expression.

Considering the wide variation between products of the same purity group, caution should be exercised in drawing conclusions concerning the immunosuppressive effects of a particular type of concentrate in haemophilia patients from study with only one product from that group.
Prevalence of thrombocytopenia in HIV‐infected and non‐HIV infected drug users and homosexual men
Authors: Mientjes G. H. C. , Ameijden E. J. C. Van , Mulder J. W. , Hoek J. A. R. Van den , Coutinho R. A. , Borne A. E. Von Dem
Published: March 2008

Summary. We studied the prevalence and risk factors for thrombocytopenia among 299 drug users and 461 homosexual men. The prevalence of thrombocytopenia was 3·3% in HIV‐negative homosexual men, 8·7% in HIV‐negative drug users, 16·4% in HIV‐positive homosexual men, and 36·9% in HIV‐positive drug users. With multivariate logistic regression HIV‐seropositivity (odds ratio 3·3), a history of injecting drugs (OR 3·9), an increased number of lymphocytes (OR 0·44), an increased number of neutrophils (OR 0·53) and a larger mean platelet volume (OR 2·8) were independently and significantly associated with thrombocytopenia. The results obtained with linear regression analysis were consistent with the results of the logistic regression. The higher prevalence of thrombocytopenia among drug users was related to a history of intravenous drug use but not to recent injecting. The mechanisms causing thrombocytopenia among HIV‐positives and HIV‐negatives seem to be related, but HIV‐infection seems to enhance thrombocytopenia in an independent way.
Lack of transcription and expression of the αIIb integrin in human early haematopoietic stem cells
Authors: Molla Annie , Andrieux Annie , Chapel Agnès , Schweitzer Annie , Berthier Rolande , Marguerie Gèrard
Published: March 2008

The glycoprotein IIb, the a subunit of the platelet integrin GPIIb‐IIIa. is a marker of megakaryocyte, but the stage of its expression during haematopoiesis remains controversial. We have examined the expression of GPIIb protein and aIIb mRNA in early human normal stem cells. We have purified stem cell expressing the CD34 surface marker (CD34+ fraction) and selected among this population quiescent cells (CD34+ MFR fraction). We have failed to detect GPIIb protein and αIIb mRNA in the pluripotential (CD34+ MFR) cells, even with polymerase chain amplification. Therefore αIIb transcription and GPIIb protein expression seemed to follow the commitment of the pluripotential cell in the mmegakaryocyte lineage.
The influence of therapeutic blocking of Gp IIb/IIIa on platelet α‐granular fibrinogen
Authors: Harrison Paul , Wilbourn Barry , Cramer Elisabeth , Faint Richard , Mackie Ian J. , Bhattacharya Shoumo , TJahiri Avijit , Tenza Danielle , Machin Samuel J. , Savidge Geoffrey F.
Published: March 2008

Recent evidence suggests that platelet a‐granule fibrinogen (fg) is derived from the plasma pool. Since platelets from patients with Type I Glanzmann's thrombasthenia (GT) are deficient in intracellular fibrinogen (fg) it was hypothesized that Gp IIb/IIIa could mediate the uptake of fg. To study the potential role of Gp IIb/IIIa in intracellular fg trafficking, the influence of therapeutic blocking of Gp IIb/IIIa on platelet fg was studied in 12 patients with stable ischaemic heart disease. Patients were either given a single intravenous dose of the monoclonal antibody 7E3 Fab (n= 4) or a combination of bolus and continuous infusion up to 24 (n= 3). 36 (n= 3) or 96 h (n= 2). All patients showed grossly prolonged bleeding times with a significant reduction of ex‐vivo ADP induced aggregation. Although, surface Gp IIb/IIIa binding sites were consistently reduced in all patients, there was a variable but delayed decrease in platelet fg relative to vWf: Ag in only six out of the 12 patients studied. The reduction in fg appeared dependent upon both dosage and duration of Gp IIb/IIIa blockade. The study provides further evidence for the novel role of Gp IIb/IIIa in the intracellular trafficking of fg to platelet and megakaryocytic alpha‐granules.
Rheological properties of rosettes formed by red blood cells parasitized by Plasmodium falciparum
Authors: Nash Gerard B. , Cooke Brian M. , Carlson Johan , Wahlgren Mats
Published: March 2008

A proportion of red blood cells parasitized by Plasmodium falciparum form rosettes with non‐parasitized red cells. Although these rosettes are thought to impair microcirculatory flow, their rheological characteristics have not been fully described. Using dual‐micropipette manipulation to pull apart individual rosettes, we found that the forces binding rosettes together were strong (average force for removal of a cell was 4.4 10‐10 N, approximately 5 times that required to detach a parasitized cell adhered to cultured endothelium). If disrupted rosettes were re‐formed, cells resetted immediately on contact, but the strength of attachment increased over minutes, and did not apparently reach its maximal level for hours. All non‐parasitized cells tested could adhere to rosette‐forming parasitized cells. Rosettes could withstand arterial flow stresses (1.4–1.6 Pa) for minutes without disintegration. To test the effects of resetting on flow resistance, the time required for entry into a 4.3 μm pipette was measured. Entry times depended strongly on the number of cells in the rosette, and averaged 35 times longer than for non‐parasitized cells. Our studies show that the cell‐cell attachments within rosettes are strong, and suggest that rosettes might survive both the arterial circulation and passage through microvessels and could contribute to the ischaemic complications of falciparum malaria.
Detection of HIV in haemopoietic progenitors
Authors: Kaczmarski R. S. , Davison F. , Blair. E. , Sutherland S. , Moxham J. , McManus T. , Mufti G. J.
Published: March 2008

Peripheral blood cytopenias present a major problem in the management of patients with HIV infection. Their pathophysiology is likely to be multifactorial. although there is controversy as to whether haemopoietic progenitors are a target for HIV. In order to investigate the haemopoietic defect in HIV infection, we looked at bone marrow culture characteristics of marrow from eight HIV+ patients compared to normal controls. We performed long‐term liquid culture (LTC) and colony forming assays for granulocytemacrophage (CFU‐GM) and granulocyte. erythroid. megakaryocyte. macrophage (CFU‐GEMM). In LTC we found normal stromal appearance and haemopoietic focus formation. There was no difference in colony assays of CFU‐GM and CFU‐GEMM between HIV+ and normal controls. Colonies taken from CFU‐GM and CFU‐GEMM were analysed for HIV DNA sequences, and we were able to detect HIV DNA in colonies from all HIV+ patients. Our results indicate that despite infection of haemopoietic progenitor cells by HIV, bone marrow function is preserved. This suggests that HIV‐related cytopenias may be due to alternative mechanisms not present in our in vitro system.
Book Reviews
Authors: Roberts B. E. , van den Berghe H.
Published: March 2008

Advances in Chronic Lymphocytic Leukaemia: Proceedings from the 5th International Workshop on Chronic Lymphocytic Leukaemia, Sitges (Barcelona), Spain, April 1991. Edited by Aaron Polliack and Emilio Montserrat

The Leukaemic Cell, 2nd edn. Edited by D. Catovsky
Lymphoid subsets and prognostic factors in multiple myeloma
Authors: Miguel J. F. San , González M. , Gascón A. , Moro J. , Hernández J. M. , Ortega F. , Jiménez R. , Guerras L. , Romero M. , Casanova F. , Sanz M.A , Sanchez J. , Portero and J.A. , Orfao A.
Published: March 2008

In a uniform series of 170 untreated myeloma patients (MM) we investigated the distribution of T cell subsets in peripheral blood (PB) and their relationship with the most relevant disease characteristics, including survival. CD4 cells were significantly decreased both in percentage and absolute numbers (P< 0.0001). On the other hand, the CD8 cells only showed a slight increase in relative numbers. Upon correlating the abnormalities in the distribution of T cells with other clinical and biological disease characteristics the most remarkable correlation was with survival. A low number of CD4 cells (< 700 × 106/1) was associated with both an advanced clinical stage and a shorter survival (20 v. 43 months,P= 0.01). Moreover, a significant correlation also exists between the decrease in CD4 cells and both high β2‐microglobulin (β2M) levels and anaemia. On the other hand, no relationship was found with the type of M‐component nor with the plasma cell phenotype. Finally multivariate analysis showed that the number of CD4 cells add independent prognostic information to other well‐established tests for the assessment of disease outcome in patients with multiple myeloma.
A new lupus anticoagulant neutralization test based on platelet‐derived vesicles
Authors: Arnout Jef , Huybrechts Erwin , Vanrusselt Marleen , Vermylen Jos
Published: March 2008

We here present an easily standardizable and reproducible procedure which clearly separates lupus anticoagulants (LA) from coagulation factor inhibitors. This new LA neutralization test makes use of platelet‐derived micro‐vesicles which were prepared as follows: gel‐filtered platelets (4 × 105/μl) were incubated with 60 μM of the calcium ionophore A23187 for 20 min at 37°C. The vesicles were separated from the platelet aggregates by centrifugation at 1000 g for 10 min. The vesicle containing supernatant was then spun down at 15000 g for 15 min, lyophilized and stored at −20°C until used. The vesicles were resuspended in plasma from normal individuals, from patients with LA activity, from patients with factor VIII inhibitors, from patients with congenital factor deficiencies and from patients receiving oral anticoagulants or intravenous heparin. A kaolin clotting time was performed in the absence (KCT) or presence of these vesicles (KCTves) and the ratios of these times to their respective mean normal times were calculated. Segregation of LA patients from all remaining patients except heparinized ones could be made with a high degree of accuracy. A thrombin time was needed to separate LA from heparinized patients. The method was highly reproducible and only minor (negligible) differences in potencies were observed between different vesicle preparations. Both the intra‐batch and the inter‐batch coefficients of variations on the KCTves were lower than 6%.
Immunological studies in HIV seronegative haemophiliacs: relationships to blood product therapy
Authors: Cuthbert R.J. G. , Ludlam C.A. , Steel C. M. , Beatson Dianne , Peutherer J. F.
Published: March 2008

Immunological studies were performed on a group of 44 haemophilia A and 15 haemophilia B patients who were treated exclusively with blood products manufactured by the Scottish National Blood Transfusion Service (SNBTS). All patients were HIV seronegative throughout the study.

Of the haemophilia A patients 14 (32%) had CD4 + lymphocyte subset counts ≤ 0.5 × 109/l, compared with one (6%) haemophilia B patient and four (8%) controls. The percentage of activated T cells was greater than 5% in 19/33 (57%) with haemophilia A, 5/9 (55%) haemophilia B and 14/50 (28%) of control subjects. β2 microglobulin values ≥2.0 mg/l were observed in 19 (43%) haemophilia A and four (26%) haemophilia B patients, compared with one (2%) control. No significant increases in serum interleukin‐2 receptor concentrations were observed in 15 haemophilia A and one haemophilia B patients. Significantly elevated levels of IgG, IgM and IgA were observed in the haemophilia A group, but elevation of immunoglobulins was restricted to the IgG class in the haemophilia B group. Of the haemophilia A patients 16/30 (53%) and 6/11 (54%) haemophilia B patients had depression of cell‐mediated immunity (CMI) as assessed by delayed‐type hypersensitivity responses to intra‐dermally injected recall antigens. There was no correlation between factor VIII or factor IX usage and changes in lymphocyte subsets, β2 microglobulin, and immunoglobulin levels. There was, however, a strong correlation between annual factor VIII usage and the degree of depression of CMI for those with haemophilia A but not for those with haemophilia B. No correlation between alterations in the immune parameters and disturbance of liver function tests was observed in either haemophilia A or haemophilia B patients.

We conclude that alloantigen or non‐HIV viral exposure due to repeated administration of factor concentrates brings about alterations in the immune response, and that these changes are more marked following exposure to intermediate purity factor VIII compared with factor IX concentrate.
Identification of an extensive ζ‐α globin gene deletion in a Chinese individual
Authors: Waye John S. , Eng Barry , Chui David H. K.
Published: March 2008

We describe a novel α‐thalassaemia‐1 deletion that removes the entire ζ‐α globin gene cluster. A Chinese couple were referred for counselling after two consecutive pregnancies ended with fetal hydrops. Gene mapping was used to demonstrate that the mother is heterozygous for the South‐east Asia α‐thalassaemia‐1 deletion (ζζα/ζζ—SEA), while the father carries an α‐thalassaemia‐1 deletion of more than 100 kilobases (ζζα/—). This newly discovered deletion extends for unknown distances 3’ and 5’ of the ζ‐α globin gene cluster and has been designated (—HW).
Short‐term study of chimaerism after bone marrow transplantation for severe aplastic anaemia
Authors: Socié G. , Landman J. , Gluckman E. , Devergie A. , Raynal B. , Esperou‐Bourdeau H. , Brison O.
Published: March 2008

Chimaerism was studied early (2 weeks to 3 months) during haematopoietic reconstitution after bone marrow transplantation in 18 severe aplastic anaemia patients (acquired SAA: 14 patients; Fanconi anaemia: four patients). Fourteen patients received marrow from an identical sibling donor, one from the phenoidentical father and three from a matched unrelated donor. Peripheral blood cell DNA was first analysed by Southern blotting with a multilocus minisatellite probe (33.6.3) or a Y chromosome specific probe (pHY2.1). For all 14 patients grafted with a genotypically identical sibling donor, the post‐graft DNA profile strictly matched the respective donor profile (minisatellite probe) or disclosed the Y chromosome specific band in the case of female patients grafted with a male donor. In contrast, for the one patient grafted in a mismatched situation and for two out of three patients grafted with a matched unrelated donor, the results indicated autologous bone marrow recovery. This difference between patients grafted with an identical sibling donor and those grafted in other situations is statistically significant (P < 0.01). The 15 patients with circulating cells of donor origin were then studied by polymerase chain reaction amplification of the DNA samples. The three male patients with a female donor were studied by amplification of a Y chromosome specific sequence (DYZ1), allowing the detection of one male cell in 104 female cells. In all three cases, residual male nucleated celts were detected. The analysis was performed by amplification of the 33.6.3 minisatellite sequence for the 12 remaining patients. No residual recipient cells were detected within the sensitivity limit of the method which is 1% in that case. This suggests that detection of residual host cells depends on the sensitivity of the technique used.
Lymphocytes from multi‐transfused patients exhibit cytotoxicity against autologous cells
Authors: Kaminski Edward R. , Hows Jill M. , Goldman John M. , Batchelor J. Richard
Published: March 2008

We previously demonstrated that multitransfused patients with severe aplastic anaemia (SAA) exhibit high numbers of alloreactive cytotoxic T lymphocyte precursors directed against their HLA identical siblings. In this study a group of patients who had received multiple blood transfusions for SAA, other haematological diseases or acute blood loss were tested for autocytotoxicity and the results compared with those of untransfused controls. These controls consisted of normal individuals, patients with chronic myeloid leukaemia (CML) or untransfused patients with SAA. There was a significantly higher degree of autocytotoxicity in multitransfused patients, than in the untransfused controls, including untransfused patients with SAA (P=0·0001). These results suggest that blood transfusion is responsible for inducing autoreactivity. In one patient, in whom both alloreactive anti‐non‐MHC and autoreactive cytotoxic T lymphocytes (CTL) had been detected, it was demonstrated that there was no crossreactivity between the alloreactive and autoreactive CTL responses. Inhibition studies using monoclonal antibodies revealed the effector cells to be T lymphocytes and the restricting determinants to be both HLA class I and II molecules.
Cytomorphology of childhood lymphoblastic leukaemia: a prospective study of 2000 patients
Authors: Lilleyman J. S. , Hann I. M. , Stevens R. F. , Richards S. M. , Eden O. B. , Chessells J. M. , Bailey C. C.
Published: March 2008

Blast cell morphology of children with lymphoblastic leukaemia (ALL) entering two national multicentre trials was prospectively reviewed by three haematologists to define the clinical importance of (a) French–American–British (FAB) classification, (b) the presence of cytoplasmic vacuoles, and (c) the presence of ‘hand‐mirror’cells.

Of 2135 evaluable children, 1907 (89%) had FAB L1 morphology and 228 (11%) L2. (L3 patients were not eligible for the trials in question). L2 patients more frequently had residual disease 14 d after starting treatment and had a significantly inferior disease‐free survival, but not if the analysis was stratified for age, sex and diagnostic white cell count (WBC), 627 (29%) had blast cells with cytoplasmic vacuoles, and showed a significant survival advantage over the remainder. Vacuoles were positively associated with a low WBC, age range 1–6 years and blast cell positivity for CD10, but their benign influence was apparent even when these variables were taken into account. ‘Hand‐mirror’(HM) cells were only studied in UKALL X, and were noted in 316/1402 (23%) children. There appeared to be an inverse correlation between HM cells and cytoplasmic vacuoles and a weak association with T‐cell immunophenotype, but no prognostic significance was evident.

FAB classification appears to be of less prognostic importance than has previously been supposed, though L2 disease is more resistant to current remission induction regimens. Hand‐mirror cells may be more common in T‐ALL, but are seen in all types and are not related to prognosis. Cytoplasmic vacuoles are predictive of a good response to current therapeutic schedules even allowing for other prognostic variables, and are the single most important morphological feature relating to prognosis in childhood ALL.
Conserved natural humoral immunity to thyroglobulin in patients with multiple myeloma
Authors: Bouanani Majida , Bataille Regis , Klein Bernard , Pau Bernard , Bastide Madeleine
Published: March 2008

We studied humoral immunity to human thyroglobulin (hTg) during the course of multiple myeloma (MM). In this report, we describe the anti‐hTg antibody activity in the sera of patients with MM. Among 63 sera tested, 28 (44%) had IgG anti‐hTg autoantibodies (aAb), 16 (25%) exhibited IgM aAb, and six (9%) had IgA anti‐hTg aAb. For the majority of sera the anti‐hTg autoantibody activity was associated with more than one immunoglobulin class. IgG anti‐hTg antibodies were observed in 9/11 patients with IgA MM and in 19/40 patients with IgG MM. The IgM anti‐hTG antibody activity was found in the sera of 11 patients with IgG MM. These results show that the anti‐hTg activity in these patients is associated with residual polyclonal immunoglobulins. However, in the serum of one patient presenting a double monoclonal gammopathy (IgG and IgA lambda MM), the anti‐hTg activity was carried by both the IgG λ and the IgA λ molecules, suggesting that in this case the activity was due to the monoclonal immunoglobulin itself. We also studied the epitopic specificity pattern of all these anti‐hTg aAb. Only three sera recognized one antigenic region on hTg. suggesting that the majority of the anti‐hTg aAb in MM patients were directed against antigenic regions other than those recognized by our panel of murine mAb. In conclusion, our results demonstrated that humoral immunity to hTg is maintained in MM patients. These data contrast with the well‐documented suppression of immunity to foreign, especially bacterial, antigens described in MM.
Philadelphia positive acute leukaemia with minor breakpoint cluster rearrangement may be a stem cell disease
Authors: Jackson G. H. , Middleton P. , Prince R. , Bown N. , Kernahan J. , Reid M. M.
Published: March 2008

The biology of Philadelphia chromosome positive (Ph+) acute lymphoblastic leukaemia (ALL) is the subject of much interest. We present a case of Ph+ ALL with a minor breakpoint cluster (mBCR) rearrangement who subsequently relapsed with Ph+ mBCR+ acute myeloid leukaemia and later with Ph+ mBCR+ acute stem cell leukaemia. This case provides further evidence that Ph+ ALL with a mBCR rearrangement may arise from a pluripotent stem cell with similar potential to that of chronic granulocytic leukaemia to undergo blastic crises with differing lineage characteristics.
Specific binding between human neutrophils and heparin
Authors: Leculier C. , Benzerara O. , Couprie N. , Francina A. , Lasne Y. , Archimbaud E. , Fiere D.
Published: March 2008

Heparin binding on polymorphonuclear leucocytes (PMNL) was characterized. Heparin binding was specific, rapid, saturable and reversible. One single class of heparin binding sites was found with a dissociation constant of 1.22 μmol/1 and 7.7×106 sites per PMNL. The binding was independent of the anticoagulant activity of heparin. Heparin affinity chromatography on radio‐iodinated cell lysates followed by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate revealed a 130 kD heparin binding protein. Heparin binding was inhibited by disodium salt of ethylenediamine tetraacetic acid. Cell surface bound heparin was functionally inactive and did not affect the inactivation of thrombin by antithrombin III. Our study demonstrates that heparin interacts with PMNL by a cell‐surface binding protein. These instructions could be consistent with the modifications of some PMNL functional properties in the presence of heparin.
Normal aggregations of glycoprotein IV (CD36)‐deficient platelets from seven healthy Japanese donors
Authors: Yamamoto Naomasa , Akamatsu Noriko , Yamazaki Hiroh , Tanoue Kenjiro
Published: March 2008

Since glycoprotein IV (GPIV) has been shown to play an important role in the interaction of platelets with collagen and thrombospondin, the aggregation and secretion of GPIV‐deficient platelets were examined. Using a binding assay with monoclonal 125I‐OKM5 antibody against CD36 antigen and crossed immunoelectrophoresis of the solubilized platelets against anti‐GPIV antibody, the platelets from seven (4.1%) out of 170 healthy Japanese donors were found to be deficient in GPIV. The GPIV‐deficient platelets showed normal aggregations in response to collagen as well as ADP, epinephrine, arachidonic acid and thrombin in comparison with GPIV‐positive platelets. Polyclonal anti‐GPIV antibody aggregated GPIV‐positive platelets but not the GPIV‐negative ones. The F(ab')2 fragments of the anti‐GPIV antibody competitively inhibited the anti‐GPIV‐induced aggregation, but did not affect the collagen‐induced aggregation of GPIV‐positive platelets. These results suggest that the deficiency of GPIV does not affect platelet aggregability.
β141 Leu is not deleted in the unstable haemoglobin Atlanta‐Coventry but is replaced by a novel amino acid of mass 129 daltons
Authors: Brennan Stephen O. , Shaw John , Allen John , George Peter M.
Published: March 2008

Reinvestigation of the structure of the β‐chain of Hb Atlanta‐Coventry (β75 Leu→Pro, β141 Leu deleted) confirmed the presence of two abnormalities; however, analysis of the aberrant βCo14 tryptic peptide by liquid secondary ion mass spectrometry indicated that the β141 Leu (mass 113 daltons) was not deleted but replaced by a novel amino acid of mass 129 daltons. The new amino acid in peptide βCo14 was uncharged at pH 6·5, more hydrophillic than leucine and susceptible to cleavage by both chymotrypsin and carboxypeptidase A. We propose that the new residue is likely to be hydroxyleucine and that it results from post‐translational oxidation of β141 Leu as a consequence of perturbation of the haem environment caused by the β75 Leu→Pro mutation in the E helix (E19). This proposal is entirely consistent with recent DNA analysis which showed that βAt‐Co was not the product of a third β‐globin gene and that neither of the two β‐globin genes. βA nor βAtlanta, contained a deletion of the β141 Leu codon. We have subsequently found this modified amino acid at position β141 in two other unstable haemoglobins, both of which involve mutations on the haem side of the E helix.
Development of a monoclonal antibody capable of differentiating platelet PLA1/PLA1, PLA1/PLA2 and PLA2/PLA2 genotypes
Authors: Lin‐Xing Liu , Nardi Michael , Flug Frances , Karpatkin Simon
Published: March 2008

A monoclonal antibody, LK‐4, has been developed which distinguishes platelet PLA1/PLA1, PLA1/PLA2 and PLA2/PLA2 genotypes on platelet glycoprotein GPIIIa of Triton‐solubilized platelet extracts. An ELISA assay has been developed which traps GPIIIa with Concanavalin A, enriching the platelet extract for the PLA antigens. A second monoclonal antibody, DEK‐10, which reacts equally with GPIIIa of PLA1/PLA1 and PLA2/PLA2 platelet extracts is employed as an internal standard to correct for individual differences in GPIIIa content, GPIIIa extracted by Triton X‐100 and GPIIIa trapped with Concanavalin A. This ELISA assay clearly differentiated 11 different PLA1/PLA1 subjects from eight PLA2/PLA2 women with a history of neonatal alloimmune thrombocytopenia as well as six unrelated obligate heterozygotes and should be useful in evaluating the PLA genotype of pregnant women and their families.
Interferon‐α therapy during pregnancy in chronic myelogenous leukaemia and hairy cell leukaemia
Authors: Baer Maria R. , Ozer Howard , Foon Kenneth A.
Published: March 2008

Interferon‐α is an effective agent in the treatment of chronic myelogenous leukaemia and hairy cell leukaemia. Because interferon‐α does not inhibit DNA synthesis, we have used it to treat these diseases during pregnancy. We report here two patients with chronic myelogenous leukaemia and two patients with hairy cell leukaemia managed with interferon‐α during pregnancy. Tnterferon‐α was well tolerated, and the pregnancies proceeded without complications. All four infants were apparently normal at birth, and all four have enjoyed normal growth and development. Interferon‐α may represent relatively safe therapy during pregnancy, because of its lack of inhibition of DNA synthesis. Nevertheless, recommendations regarding the use of interferon‐α to treat chronic leukaemias during pregnancy must await further experience with this agent in this setting.
A randomized study of intermediate versus conventional‐dose cytarabine as intensive induction for acute myelogenous leukaemia
Authors: Schiller Gary , Gajewski James , Nimer Stephen , Territo Mary , Ho Winston , Lee Myung , Champlin Richard
Published: March 2008

The optimal dose of cytarabine for induction chemotherapy is unknown. Most studies have utilized doses of 100–200 mg/m2/d, although higher doses have been proposed to increase the concentration of the active metabolite ara‐CTP within leukaemia cells. To address this question 101 adults with newly diagnosed acute myeloid leukaemia were randomized to receive treatment with daunorubicin and either conventional‐dose cytarabine (200 mg/m2/d by continuous infusion) or an intermediate‐dose of cytarabine (500 mg/m2 every 12 h). 36/51 (71%) patients assigned to conventional‐dose cytarabine achieved complete remission compared to 37/50 (74%) who achieved remission with intermediate‐dose cytarabine (P = 0.9). Patient age significantly affected remission rate. 8/17 patients age >60 assigned to conventional‐dose cytarabine and 10/17 assigned to intermediate‐dose cytarabine achieved complete remission compared to 27/33 patients under age 60 assigned to the conventional dose and 28/34 patients assigned to the intermediate dose arm (P=0.004). Actuarial 4‐year disease‐free survival for patients assigned to conventional‐dose cytarabine was 20.16% versus 28.17% for patients assigned to intermediate‐dose cytarabine (P=0–9). We conclude that intermediate dose cytarabine did not substantially improve results of induction chemotherapy for acute myeloid leukaemia.
The immunophenotype in infant acute lymphoblastic leukaemia: correlation with clinical outcome. An Italian multicentre study (AIEOP)
Authors: Basso G. , Putti M. C. , Cantú‐Rajnoldi A. , Saitta M. , Santostasi T. , Santoro N. , Lippi A. , Comelli A. , Felici L. , Favre C. , Mancuso G. Russo , Guglielmi C. , Paolucci P. , Biondi A. , Rondelli R. , Pession A.
Published: March 2008

A detailed analysis of immunophenotype of 112 infants aged less than 18 months with acute lymphoblastic leukaemia (ALL) was performed. Patients were divided into three groups on the basis of age at presentation (under 6 months: group 1; 6–12 months: group 2; 13–18 months: group 3). There were three cases of T‐ALL (2.6%). The proportion of other subtypes was: common ALL in 59 patients (52.68%), pre‐B ALL in 15 patients (13.3%), pre‐pre‐B ALL in 27 (24.1%) and acute undifferentiated leukaemia (AUL) in eight patients (7.14%).

In non‐T ALL, positivity to CD10 (corresponding to C‐ALL and pre‐B ALL) was distributed in the three age groups as follows: 38.88% (group I) 65.38% (group II) and 86.36% (group III). Conversely, immature phenotypes (pre‐pre‐B and AUL) were found more often in the younger patients of groups I and II, as well as anomalous phenotypes, such as the presence of myeloid antigens (MyAg) and of CD7. Prognostic significance was evaluated as event‐free survival (EFS) by statistical analysis. A better outcome in CD10‐positive ALL than in CD01‐negative ones (48% v. 25% of long‐term survivors) was demonstrated in all infants. Similarly, EFS was significantly better in MyAg‐negative than in MyAg‐positive cases. These results were confirmed also when adjusting for white blood cell count. This allowed the identification of CD10‐negative, MyAg‐positive ALL, which were relatively more frequent in infants and had a poorer clinical outcome with the current therapies. This study stresses the prognostic relevance of the immunological study in infant leukaemias and its utility in choosing different therapeutic modalities for poor risk phenotypes.
Mini‐BEAM followed by BEAM and ABMT for very poor risk Hodgkin's disease
Authors: Chopra R. , Linch D. C. , McMillan A. K. , Blair S. , Patterson K. G. , Moir D. , Richards J. D. M. , Cervi P. , Kinsey S. , Goldstone A. H.
Published: March 2008

Summary. High dose chemotherapy and autologous bone marrow transplantation (ABMT) is an effective form of salvage therapy in patients with relapsed or resistant Hodgkin's disease. Patients with large tumour masses at the time of ABMT have a poorer prognosis and we have therefore administered intermediate dose BCNU. etoposide, cytarabine and melphalan (mini‐BEAM) prior to high dose therapy with the same agents (BEAM) and ABMT in such patients. In addition we have used the same strategy in patients with bone marrow infiltration at the time of relapse in an attempt to clear the bone marrow for transplant. A total of 23 patients received mini‐BEAM and 21 proceeded to BEAM and ABMT. Platelet engraftment was delayed compared to BEAM recipients who had not received mini‐BEAM (P= 0.008) but there was only one procedure related death. Responses to BEAM and ABMT were not predicted by the response to mini‐BEAM indicating a dose response effect at the upper end of the dose intensity spectrum. At 2 years, the overall survival and progression free survival are 61% and 46% respectively for this group of Hodgkin's patients with extremely poor prognosis.
The relationship between increased platelet count and megakaryocyte size in bronchial carcinoma
Authors: Kristensen S. D. , Bath P. M. W. , Gladwin A.‐M. , Martin J. F.
Published: March 2008

This study investigates megakaryocyte size in bronchial carcinoma and its relationship to platelet count, mean platelet volume and platelet function, the latter determined in vivo by measurement of the cutaneous bleeding time. Measurements were performed in 19 patients with bronchial carcinoma and 11 patients with a hiatus hernia (controls). Platelet count (335.123 v. 241 × 72.109/1) and megakaryocyte total area (772.135 v. 666.105μm2) (mean × standard deviation) were both significantly increased in patients with bronchial carcinoma when compared to controls whilst the bleeding time was shorter in the patient group (253.116 v. 321.80 s). Patients with bronchial carcinoma were older (60.2 ×8.4 v. 48.8 ×13.2 years) and smoked more heavily (760.511 v. 92.175 cigarette years) than controls. Megakaryocyte size, platelet count and bleeding time all correlated with smoking habit (r=0.395, P=0.031; r= 0.622, P<0.001; r=‐0.515, P=0.04 respectively). There were no statistically significant differences between the groups with respect to gender or mean platelet volume. When considering all patients, significant correlations existed between platelet count and megakaryocyte area (r= 0.400, P= 0.029), and bleeding time and megakaryocyte area (r=‐0.365, P= 0.047). The megakaryocyte nuclear and cytoplasmic areas were positively correlated (r=0–855, P<0.001). This study suggests that the elevated platelet count in patients with bronchial carcinoma is mediated by an increase in the size of bone marrow megakaryocytes although the results are compounded by the association between smoking and megakaryocyte/platelet parameters.
Factor VIII gene deletions in haemophilia A patients in Czechoslovakia
Authors: Křepelová Anna , Vorlová Zdena , Zavadil Jiŕí , Brdička Radim
Published: March 2008

Genomic DNA from 90 Czechoslovak haemophilia A patients from 81 pedigrees was analysed by Southern blotting and hybridization with factor VIII cDNA probes. Three partial deletions of the factor VIII gene were identified and characterized: a 4.8 kilobase (kb) deletion eliminating exon 10 in one patient with severe haemophilia A without inhibitor, a 6.1 kb deletion eliminating the 3’ part of intron 13 and the 5’ part of exon 14 in two related severe haemophiliacs, but only one of them produced inhibitor, and a 4.6 kb deletion eliminating the 3’ part of intron 13 and the 5’ part of exon 14 in a severe haemophiliac with high‐titre inhibitor.

Besides these three deletions, three different restriction site variants without apparent loss of DNA sequence were found.
Expression of adhesion molecules LFA‐3 and N‐CAM on normal and malignant human plasma cells
Authors: Barker H. F. , Hamilton M. S. , Ball J. , Drew M. , Franklin I. M.
Published: March 2008

Normal and malignant plasma cells were investigated for the expression of seven cellular adhesion molecules by immunofluorescence microscopy. The antigens investigated were CD2 and its ligand, LFA‐3 (CD58), LFA‐1α (CD11a) and LFA‐1β (CD18) and their ligand ICAM‐1 (CD54), H‐CAM (lymphocyte homing receptor; CD44) and N‐CAM (CD56). Marrow from 18 patients with myeloma, two with plasma cell leukaemia (PCL), four with monoclonal gammopathy of uncertain significance (MGUS) and 10 normal allogeneic bone marrow donors was studied. All plasma cells from normals and multiple myeloma patients were negative for CD2, CD11a and CD18. All normal and myeloma marrow plasma cells were positive for ICAM‐1. 16/18 myeloma cases tested, and all other samples (normal, MGUS and PCL), contained plasma cells positive for H‐CAM. Only one normal, but 12/16 myelomas tested were positive for N‐CAM (P<0.02). One of four MGUS cases was moderately positive and one other weakly positive for N‐CAM. Both PCLs were N‐CAM negative. 12/18 myelomas were positive for LFA‐3, but only two normals (P<0.05). All MGUS cases were negative for LFA‐3, as was one PCL, the other being weakly positive. Three cases were negative for both adhesion molecules, three cases expressed only N‐CAM or LFA‐3 and 10 cases expressed both. LFA‐3 and N‐CAM are expressed significantly in myeloma rather than normal plasma cells. Cases of MGUS may express N‐CAM but not, in this small series, LFA‐3. Plasma cells in the peripheral blood (PCL) and plasma cell lines express little or no LFA‐3 or N‐CAM.
Detection of allele‐specific expression of N‐RAS oncogenes in human leukaemia cells
Authors: Lübbert Michael , Mertelsmann Roland , Herrmann Friedhelm
Published: March 2008

We report analysis of allele‐specific expression of N‐RAS transcripts in myeloid leukaemic cells and cell lines. Expression was assessed by an assay of reverse‐transcription/PCR combined with differential hybridization with mutation‐specific oligonucleotides. In cells from all patients with acute myeloid leukaemia (AML) and in myeloid cell lines HL‐60 and THP‐1, expression of both the wild‐type allele and the abnormal allele altered by a point mutation could be detected, albeit not always at comparable levels. This might be due for instance to allelic exclusion. The assay described provides a means of analysing the degree of expression of dominant oncogenes.
HLA‐identical sibling donor bone marrow transplantation for chronic myeloid leukaemia in first chronic phase: influence of GVHD prophylaxis on outcome
Authors: Marks David I. , Hughes Timothy P. , Szydlo Richard , Kelly Steven , Cullis Jonathan O. , Schwarer Anthony P. , Mackinnon Stephen , Apperley Jane , Barrett A. John , Hows Jill M. , Goldman John M.
Published: March 2008

We have analysed the results of treating 140 consecutive patients with chronic myeloid leukaemia (CML) in chronic phase by bone marrow transplantation (BMT) using marrow from HLA‐identical siblings performed between February 1981 and July 1991. Three different regimens were used sequentially to prevent graft‐versus‐host disease (GVHD): cyclosporin A (CsA) alone (n= 39), T‐cell depletion of donor marrow (n= 51) and CsA with methotrexate (MTX) (n= 50). Eighty‐four patients (61%) survive at a median of 49 months from BMT (range 3–120). The actuarial overall and leukaemia‐free survivals at 5 years were 52% and 41% respectively. The actuarial probabilities of leukaemia‐free survival and haematological relapse at 2 years for the CsA only group were 65% and 4%, for the T‐cell depletion group 40% and 41% and for the CsA/MTX group 68% and 6% respectively. For the T‐cell depletion group the probability of leukaemia‐free survival was significantly lower (P < 0.001) and the probability of relapse significantly higher (P<0·001) than for other methods of GVHD prophylaxis: differences between the other two groups were not significant. Previous reports that T‐cell depletion with Campath‐1M results in a high rate of relapse are confirmed. Patients in the CsA/MTX group have been monitored with cytogenetic and polymerase chain reaction studies for residual BCR/ABL transcripts. We conclude that the combination of CsA/MTX is currently the best available approach to prevention of GVHD after BMT for CML and in our hands it is not associated with a major risk of relapse.