bluuebunny/crossref_metadata_2025 huggingface dataset:
Increase in Vδ1+γδ T cells in the peripheral blood and bone marrow as a selective feature of HIV‐1 but not other virus infections
Authors: Rossol Rita , Dobmeyer JÜrgen M. , Dobmeyer Thomas S. , Klein Stefan A. , Rossol Siegbert , Wesch Daniela , Hoelzer Dieter , Kabelitz Dieter , Helm Eilke B.
Published: March 2003

Dysregulation of T‐cell receptor (TCR) αβ bearing lymphocytes and an increase in Vδ1+γδ T cells are typical features of HIV‐1 infection. However, the role of γδ T cells remains unclear. Therefore, peripheral blood mononuclear cells (PBMC) of 103 HIV‐1‐infected patients were investigated with respect to expression of Vδ1. These results were compared to the Vδ1 expression of bone marrow mononuclear cells (BMMC). In contrast to healthy controls, Vδ1+ cells dominated among both PBMC and BMMC in HIV‐1‐infected patients. Analysis of the coexpression of CD25, CD8, HLA‐DR and CD45RO revealed a high prevalence of Vδ1/CD45RO and Vδ1/HLA‐DR double‐positive PBMC only in HIV‐1‐infected patients but not in healthy donors. Furthermore, analysis of the γδ TCR repertoire in patients infected with hepatitis B virus, hepatitis C virus, herpes simplex virus (HSV)‐1 and HSV‐2 showed that the selective enhancement of Vδ1+ cells was restricted to HIV infection and not observed in other virus diseases. Our data provide further support for the involvement of γδ T cells in immunosuppression and progression of HIV infection.
Progenitor cell subsets and engraftment kinetics in children undergoing autologous peripheral blood stem cell transplantation
Authors: Gonzalez‐Requejo Angel , Madero Luis , DÍaz Miguel A. , Villa Marta , GarcÍA‐Escribano Carmen , Balas Antonio , Lillo Rosa , GarcÍA‐Sanchez Félix , Benito Ana , Vicario Jose L.
Published: March 2003

The main objective of the present study was to determine the role of CD34+ cell subsets in the haemopoietic recovery of children undergoing peripheral blood stem cell transplantation. For this purpose, 38 leukaphereses from 33 children with malignancies mobilized with G‐CSF were analysed. Using dual‐colour flow cytometry, different subpopulations of CD34+ cells were quantified and the number of each reinfused subsets correlated with haemopoietic resurgence. Multivariate analysis showed that the number of CD34+CD38− cells and CD34+CD38+ cells correlated better with time to neutrophil and platelet recovery, respectively, than the total number of CD34+ cells. Threshold values for rapid haemopoietic recovery, determined by the receiver operating characteristic analysis, were found to be 0.5 × 106 CD34+CD38− cells for neutrophil engraftment, and 2.0 × 106 CD34+CD38+ cells for platelet recovery. It is suggested that the analysis of CD34+ cell subsets could increase understanding of the repopulation capacity of a given leukapheresis product in peripheral blood stem cell transplantation procedures in children. In particular, this procedure could be extremely useful when low numbers of CD34+ cells are collected.
Interferon α activates the tyrosine kinase Lyn in haemopoietic cells
Authors: Uddin Shahab , Grumbach Isabella M. , YI Taolin , Colamonici Oscar R. , Platanias Leonidas C.
Published: March 2003

We investigated whether the src‐family tyrosine kinase Lyn is involved in the generation of interferon α (IFNα) signals in haemopoietic cells. In vitro kinase assays using IFNα‐sensitive cells of B‐cell origin demonstrated the presence of IFNα‐dependent kinase activity in anti‐Lyn immunoprecipitates. Further studies demonstrated that Lyn associates via its src homology 2 (SH2) domain with the Janus family tyrosine kinase Tyk‐2. This interaction was IFNα‐dependent and involved direct binding of the SH2 domain of Lyn to the IFNα‐activated form of Tyk‐2. Thus, during binding of IFNα to its receptor in malignant haemopoietic cells, Lyn is engaged in an IFNα‐signalling pathway, probably downstream of Tyk‐2.
Successful use of protease inhibitors in HIV‐infected haemophilia patients
Authors: Merry Concepta , Mcmahon Corrina , Ryan Mairin , O'shea  , Mulcahy Fiona , Smith Owen P.
Published: March 2003

The haemophilias are a group of inherited haemostatic disorders that require regular clotting factor replacement therapy in the severe and moderately severe subgroups. Prior to the introduction of adequate viral inactivation methods in 1985, haemophilia patients were at exceptionally high risk of contracting blood‐borne viruses from factor concentrates as each batch was derived from the plasma of thousands of donors. As a result, approximately 60% of these patients were infected with HIV between 1979 and 1985, and HIV infection now contributes significantly to the morbidity and mortality seen in this group.

Protease inhibitors (PIs) have been shown to significantly log reduce viral loads and increase CD4 cell counts in HIV‐infected individuals. Recently, there has been concern about their use in HIV‐infected haemophilia patients following increased bleeding episodes in some patients during PI therapy. We prospectively studied the effect of PI therapy in 20 HIV‐infected haemophilia patients at our centre over a 6‐month period. The mean increase in CD4 cell count was 70 × 106/l and the mean log decrease in viral load was 1.59 over the study period. Gastrointestinal side‐effects (nausea and vomiting in five, diarrhoea in two) were the most frequent and resulted in discontinuation of the medication in two patients. Factor concentrate usage for the group on and off study was similar. One severe FVIII patient reported a single episode of an unusual bleed which responded promptly to FVIII concentrate infusion. The significant clinical and laboratory benefits in terms of HIV disease and the paucity of added bleeding complications suggest that PI therapy should not be withheld from HIV‐infected haemophilics. Further prospective studies evaluating the efficacy and possible haemostatic complications related to these promising inhibitors of the HIV protease are needed.
Significance of trilineage myelodysplasia in de novo acute myeloid leukaemia during remission rather than at diagnosis
Authors: Tamura Shu , Takemoto Yoshinobu , Wada Hiroshi , Itoh Tohru , Mori Ako , Saheki Kaname , Okada Masaya , Takatsuka Hiroyuki , Fujimori Yoshihiro , Okamoto Takahiro , Kakishita Eizo
Published: March 2003

Patients with trilineage myelodysplasia (TMDS) in de novo acute myeloid leukaemia (AML) at diagnosis and remission were clinically evaluated between 1983 and 1996. AML with TMDS (AML/TMDS) was observed in 20 (12%) of 162 patients with de novo AML at diagnosis. Complete remission (CR) was achieved with combination chemotherapy in 12 (67%) of 18 AML/TMDS cases. This CR rate was relatively worse than the rate of 78% (106/136 cases) of AML without TMDS, but this difference was not significant. Disease‐free survival curves also showed no difference between AML/TMDS and AML without TMDS. During remission, eight (67%) of 12 AML/TMDS cases had myelodysplastic remission marrow (AML/MRM). AML/MRM was also seen in seven (7%) of 106 AML cases without TMDS. The actuarial disease‐free survival was significantly lower in AML/MRM than in AML without MRM (P = 0.0003). All of the AML/MRM cases exhibited early leukaemic relapse; median remission duration was only 9 months. Clonal changes occurred in two cases of AML/TMDS and five cases of AML/MRM at the time of relapse. These findings suggest that TMDS during remission predicts a poorer prognosis and early leukaemic relapse when compared with the absence of TMDS.
Associations of protein C and protein S with serum lipid concentrations
Authors: Maccallum  , Cooper  , Martin  , Howarth  , Meade  , Miller
Published: March 2003

Difficulties in the laboratory measurement of protein C and protein S levels cause problems in the diagnosis of deficiency states in individual patients and may complicate estimation of the prevalence of these states in the general population. Some difficulties may be due to unappreciated influences affecting the measured levels of proteins C and S. We measured protein C activity and antigen, total and free protein S antigen, and serum total cholesterol, high‐density cholesterol and triglyceride in a community‐based study of 150 adults (73 male, 77 female), age range 23–80 years. Participants were identified from the list of a single general practice by stratified random sampling within sex and decade of age. Protein C activity and antigen were strongly associated with serum lipids, mean levels increasing by approximately 0.25 u/ml as total cholesterol and triglyceride concentration each rose from the 5th to 95th centile. Total protein S antigen concentration was associated with total cholesterol, the mean rising by over 0.1 u/ml as total cholesterol increased from the 5th to the 95th centile, whilst a similar rise in triglyceride was associated with an increase in mean free protein S of more than 0.3 u/ml. Overall, physiological variation in total cholesterol and triglyceride concentration was associated with significant variation in protein C and protein S levels, independent of age and sex, suggesting that it is important to take serum lipids into account when investigating patients for protein C or protein S deficiency. Failure to do so may be misleading in some circumstances.
Immunophenotype of adult and childhood acute promyelocytic leukaemia: correlation with morphology, type of PML gene breakpoint and clinical outcome. A cooperative Italian study on 196 cases
Authors: Guglielmi Cesare , Paola Martelli Maria , Diverio Daniela , Fenu Susanna , Luce Vegna Maria , CantÙ‐Rajnoldi Angelo , Biondi Andrea , Grazia Cocito Maria , Del Vecchio Luigi , Tabilio Antonio , Avvisati Giuseppe , Basso Giuseppe , LO Coco Francesco
Published: March 2003

Acute promyelocytic leukaemia (APL), characterized by a specific PML‐RARα fusion gene resulting from translocation t(15;17) and by a high response rate to differentiation therapy with all‐trans retinoic acid, presents clinical (varying WBC counts, age and treatment outcome), morphological (hypergranular M3 and hypogranular M3V) and molecular (three isoforms of PML breakpoint) heterogeneity.

We correlated leukaemic immunophenotype with these aspects in 196 molecularly confirmed APLs (63 children and 133 adults) in Italy. The bcr3 isoform (P = 0.05) and FAB M3V (P = 0.05) were more frequent in children. We confirmed in APL an immunophenotype characterized by frequent expression of CD13, CD33 and CD9 and rare expression of HLA‐DR, CD10, CD7 and CD11b. However, we recognized CD2 in 28%, CD34 in 23% and CD19 in 11% of cases and demonstrated by double labelling that CD34 and CD2 may be co‐expressed. CD2, CD34 and CD19 were significantly intercorrelated, and variably associated to other features: CD2 and CD34 with PML bcr3 (P < 0.001 and P < 0.001, respectively) and with M3V (P < 0.001 and P = 0.002), whereas only CD19 was directly correlated with WBC counts and only CD2 positively influenced CR rate (logistic model) and event‐free survival (Cox model). We conclude that immunophenotype plays a role in the determination of the biological and clinical heterogeneity of childhood and adult APL.
Decreased activity of the multidrug resistance P‐glycoprotein in acquired aplastic anaemia: possible pathophysiologic implications
Authors: Calado Rodrigo T. , Garcia Aglair B. , FalcÃO Roberto P.
Published: March 2003

To address a possible impairment of multidrug resistance mechanisms in acquired aplastic anaemia (AA), the functions of P‐glycoprotein (P‐gp) and multidrug resistance‐associated protein (MRP) were respectively assessed by rhodamine 123 (Rh123) and daunorubicin (DNR) efflux in peripheral blood lymphocytes from AA patients. The proportion of Rh123‐effluxing T cells was significantly decreased in AA, relative to controls. Interestingly, these changes were also present in patients with AA in remission. Conversely, Rh123 efflux in B and natural killer (NK) cells and DNR efflux in peripheral blood lymphocytes were unchanged. These data indicated that P‐gp activity was decreased in AA not only during the development of the disease, but also after remission, introducing a new concept on the pathophysiology of AA by suggesting that it may contribute to drug‐induced injury to haemopoietic cells in some cases of AA, by increasing the proportion of susceptible cells.
A genetic syndrome associating dehydrated hereditary stomatocytosis, pseudohyperkalaemia and perinatal oedema
Authors: Grootenboer  , Schischmanoff  , Cynober  , Rodrigue  , Delaunay  , Tchernia  , Dommergues
Published: March 2003

Dehydrated hereditary stomatocytosis is a haemolytic anaemia with an underlying impairment of monovalent cation transport. It is sometimes associated with pseudohyperkalaemia (e.g. an increase of kalaemia when blood is left at room temperature) or with perinatal ascites. We report a case in which dehydrated hereditary stomatocytosis, pseudohyperkalaemia and perinatal oedema coexisted, and were transmitted en bloc in a dominant fashion. Transfusions did not cure the oedema, that spontaneously receded after a few months. We assume that the various manifestations stemmed from one single altered locus, yet to be determined.
All‐trans‐retinoic acid up‐regulates CD38 but not c‐Kit antigens on human marrow CD34+ cells without recruitment into cell cycle
Authors: Herault Olivier , Domenech Jorge , Degenne Michel , Bremond Jean‐Louis , Sensebe Luc , Bernard Marie‐Christine , Binet Christian , Colombat Philippe
Published: March 2003

Retinoids, especially all‐trans‐retinoic acid (ATRA), are well known for their differentiating activity on HL‐60 cells. Moreover ATRA induces CD38 antigen overexpression on these cells. In this study we examined the effects of ATRA on purified normal CD34+ cells from adult human marrows incubated with ATRA (1 μM) or stem cell factor (SCF) after 7 d liquid cultures in serum‐deprived medium. Before and after the incubation, CD34+ cells were studied by flow cytometry to evaluate the cell‐surface expression of CD38 and c‐Kit antigens and the cycle status of these cells using high‐resolution analysis (DNA content v Ki‐67 antigen expression) to clarify the functional meaning of antigenic variations. When compared with control cultures, ATRA‐treated cells displayed changes in their immunophenotypic profile. Particularly relevant was the up‐regulation of CD38 antigen with a mean (±SEM) fold increase of 2.1 ± 0.1 (P = 0.028) for geometric mean fluorescence intensity (GMFI), without modulation of c‐Kit expression. SCF only down‐regulated expression of c‐Kit with a fold decrease of 4.6 ± 0.9 for GMFI (P = 0.043). Unlike SCF, ATRA did not induce CD34+ cells to entry into cell cycle despite increased levels of surface CD38 antigen. Moreover morphological and functional assays did not argue for an ATRA‐induced maturation process. Contrary to steady‐state cells, CD34+ cells treated with pharmacological doses of ATRA alone displayed CD38 over‐expression without change in c‐Kit levels and cycle status, suggesting an absence of maturation pressure.
Identification of new type 2B von Willebrand disease mutations: Arg543Gln, Arg545Pro and Arg578Leu
Authors: Hilbert L. , Gaucher C. , Abgrall J. F. , Parquet A. , Trzeciak C. , Mazurier C.
Published: March 2003

We report the identification in five patients (three families) affected with type 2B von Willebrand disease (VWD) of three heterozygous nucleotide substitutions at the codon for arginine 543, 545 and 578 of the mature von Willebrand factor (VWF) subunit resulting in a glutamine, proline and leucine substitution, respectively. These mutations are located in the A1 loop where prevalent type 2B mutations (Arg543Trp, Arg545Cys and Arg578Gln) have been already identified at the same positions. By in vitro mutagenesis of full‐length cDNA of VWF and transient expression in Cos‐7 cells, we have shown that the six corresponding mutated recombinant VWFs (Gln543, Trp543, Cys545, Pro545, Leu578 and Gln578 rVWF) exhibited quantitatively normal expression and normal multimeric pattern but increased ristocetin‐ and botrocetin‐induced binding to platelets as compared with that for wild‐type rVWF. The two mutations at position 545 induced the greatest reactivity for GPIb of corresponding rVWFs as compared to the two mutations at positions 543 and 578.
CD40 regulation of death domains containing receptors and their ligands on lymphoma B cells
Authors: Ribeiro Patricia , Renard Nathalie , Warzocha Krzysztof , Charlot Carole , Jeandenant Lorena , Callet‐Bauchu Evelyne , Coiffier Bertrand , Salles Gilles
Published: March 2003

Within the tumour necrosis factor (TNF) family the induction of apoptosis is restricted to some ligand–receptors pairs, including TNF–TNF receptor type I (TNFRI/p55), FasL‐Fas, TNF‐related apoptosis‐inducing ligand (TRAIL) and its death‐receptors (DR)‐4 and ‐5. The pair CD40L–CD40 belongs to the same family but rescues B cells from apoptosis. To investigate how these opposing actions are cross‐linked, purified follicular lymphoma (FL) cells were activated upon a human CD40L‐transfected murine fibroblastic layer, then RNA messengers for the above molecules were analysed using RT‐PCR. The observed down‐modulation of TRAIL and up‐regulation of TNF and Fas transcripts might account for CD40‐CD40L‐mediated FL cell survival.
Evaluation of cardiac status in iron‐loaded thalassaemia patients following bone marrow transplantation: improvement in cardiac function during reduction in body iron burden
Authors: Mariotti Egidio , Angelucci Emanuele , Agostini Alberto , Baronciani Donatella , Sgarbi Ernesto , Lucarelli Guido
Published: March 2003

Iron‐induced cardiac disease is the primary cause of death in transfused patients with thalassaemia major. The beneficial effects of deferoxamine mesylate on clinical cardiac disease have been well described but the impact of therapy on subclinical cardiac dysfunction is unknown. To assess the reversibility of subclinical cardiac dysfunction we studied the cardiac status during iron depletion treatment (phlebotomy) in iron overloaded patients, cured of thalassaemia by marrow transplantation, without clinical manifestation of heart failure but with alteration in both left ventricular diastolic function and in contractility property. 32 patients were studied and demonstrated a slight but significant impairment in the morphology and function if compared with matched normal controls. 17 of these patients were submitted to sequential echocardiographic evaluations during the phlebotomy programme. Following completion of the programme, normalization of the indices of contractility and normalization of diastolic function were observed. This study indicates that transplanted thalassaemia patients with subclinical left ventricular diastolic dysfunction and impaired left ventricular contractility may reverse these processes with an effective regimen of iron reduction such as phlebotomy.
IL4 production and increased CD30 expression by a unique CD8+ T‐cell subset in B‐cell chronic lymphocytic leukaemia
Authors: De TOTERO DANIELA , REATO GIGLIOLA , MAURO FRANCESCA , CIGNETTI ALESSANDRO , FERRINI SILVANO , GUARINI ANNA , GOBBI MARCO , GROSSI CARLOENRICO , FOA ROBERT
Published: March 2003

Phenotypic and functional abnormalities within the residual non‐B‐cell compartment of B‐cell chronic lymphocytic leukaemia (CLL) suggest an interaction between tumour cells and host immune effectors. To explore the possibility of a polarized Th1/Th2 response we have studied CD30 antigen expression and the pattern of cytokine production by purified CLL T cells. Activated T cells from CLL patients showed a significant increase in the expression of CD30 compared to normal controls. Accordingly, high levels of soluble CD30 were detected in supernatants from activated T‐cell cultures, as well as in CLL serum samples. Messenger RNA for IL4 was found in both resting and, to a greater extent, in activated CLL T lymphocytes. The latter cells were also capable of releasing IL4. Three‐colour immunofluorescence analyses revealed a strong CD30 expression in the CD3+/CD8+/CD28− large granular lymphocyte subset, which is considerably expanded in CLL. Production of IL4, as well as expression and release of CD30 by these T cells, was conclusively demonstrated at the clonal level. These findings document an expansion of a peculiar subset of ‘Th2‐like’ cells in CLL, with an increased IL4 production and CD30 expression and release, that are likely to contribute to both the B‐cell accumulation and immune‐defects characteristic of this disease.
Mutation at position −12 of intron 45 (c → t) plays a prevalent role in the partial skipping of exon 46 from the transcript of allele αLELY in erythroid cells
Authors: Wilmotte Rick , Marechal Joëlle , Delaunay Jean
Published: March 2003

Allele αLELY is a common low‐expression allele of the erythroid spectrin SPTA1 gene. It results in the aggravated expression of hereditary elliptocytosis due to SPTA1 gene mutations occurring in trans. Allele αLELY contains, in particular, mutations in introns 45 and 46, both in polypyrimidine tracts, and causes the partial skipping of exon 46. The corresponding six amino acids belong to the nucleation site where the dimerization process of spectrin begins. In this work we investigated the cause of exon 46 partial skipping. We made four types of constructs with or without the intron 45 mutation, and with or without intron 46 mutation. Intron 45 mutation by itself dramatically triggered partial skipping of exon 46. Intron 46 mutation had no effect by itself. It was not possible to assess whether it modulated, even to a very small extent, the activity of intron 45 mutation. Taken together, intron 45 mutation is the prevalent, if not the exclusive, determinant of the partial skipping of exon 46 in the transcript of allele αLELY.
The role of haematological factors in diabetic peripheral arterial disease: the Edinburgh Artery Study
Authors: Lee Amanda J. , MacGregor Andrew S. , Hau Cathryn M. , Price Jacqueline F. , Rumley Ann , Lowe Gordon D. O. , Fowkes F. Gerald R.
Published: March 2003

The relationship between haematological factors and peripheral arterial disease (PAD) among diabetics has not been widely examined. 1592 men and women aged 55–74 years were selected from the general population. They underwent an assessment for PAD and a glucose tolerance test. 288 subjects (18.7%) were identified as having diabetes or impaired glucose tolerance (IGT). Among the diabetes/IGT group, median levels of fibrinogen, von Willebrand factor (VWF), tissue plasminogen activator (t‐PA), fibrin D‐dimer and plasma viscosity were higher in subjects with PAD than those without PAD (P ≤ 0.05). The prevalence of PAD was higher in those with diabetes/IGT (20.6%) compared to those with normal glucose tolerance (12.5%) (odds ratio 1.64; 95% CI 1.17, 2.31). After separate adjustment for fibrinogen, VWF, t‐PA, fibrin D‐dimer, leucocyte elastase, plasma viscosity and haematocrit, those with diabetes/IGT no longer had a significantly higher risk of PAD compared to those with a normal glucose tolerance test. Simultaneous adjustment for the first four of these haematological factors reduced the risk of PAD among subjects with diabetes/IGT to 1.11 (95% CI 0.76, 1.63). Increased levels of haemostatic factors may partly explain the higher prevalence of PAD in diabetic/IGT subjects compared to normal glucose‐tolerant subjects. Future randomized controlled trials involving the indirect lowering of levels of haematological factors should help to explain whether the associations reported here are of causal significance.
Peripheral blood progenitor cell (PBPC) counts during steady‐state haemopoiesis enable the estimation of the yield of mobilized PBPC after granulocyte colony‐stimulating factor supported cytotoxic chemotherapy: an update on 100 patients
Authors: Fruehauf Stefan , Schmitt Karin , Veldwijk Marlon R. , Topaly Julian , Benner Axel , Zeller W. Jens , Ho Anthony D. , Haas Rainer
Published: March 2003

Peripheral blood progenitor cells (PBPC) can be mobilized using chemotherapy and granulocyte colony‐stimulating factor (G‐CSF). We and others previously reported a correlation of steady‐state PBPC counts and the PBPC yield during mobilization in a small group of patients. Here we present data on 100 patients (patients: 25 non‐Hodgkin's lymphoma (NHL), five Hodgkin's disease, 35 multiple myeloma (MM), 35 solid tumour) which enabled a detailed analysis of determinants of steady‐state PBPC levels and of mobilization efficiency in patient subgroups. Previous irradiation (P = 0.0034) or previous chemotherapy in patients with haematological malignancies (P = 0.0062) led to a depletion of steady‐state PB CD34+ cells. A correlation analysis showed steady‐state PB CD34+ cells (all patients: r = 0.52, P < 0.0001; NHL patients, r = 0.69, P = 0.0003; MM patients: r = 0.66, P = 0.0001) and PB colony‐forming cells can reliably assess the CD34+ cell yield in mobilized PB. In patients with solid tumour a similar trend was observed in mobilization after the first chemotherapy cycle (r = 0.51, P = 0.05) but not if mobilization occurred after the second or further cycle of a sequential dose‐intensified G‐CSF‐supported chemotherapy regimen, when premobilization CD34+ counts were 18‐fold elevated (P = 0.004). When the patients with MM (r = 0.63, P = 0.0008) or with NHL (r = 0.65, P = 0.006) were analysed separately, a highly significant correlation of the steady‐state PB CD34+ cell count to the mean leukapheresis CD34+ cell yield was found, whereas no correlation was observed for patients with a solid tumour. For patients with haematological malignancies estimates could be calculated which, at a specific steady‐state PB CD34+ cell count, could predict with a 95% probability a defined minimum progenitor cell yield. These results enable recognition of patients who mobilize PBPC poorly and may assist selection of patients for novel mobilization regimens.
Clinical significance of cyclin‐dependent kinase inhibitor p27Kip1 expression and proliferation in non‐Hodgkin's lymphoma: independent prognostic value of p27Kip1
Authors: Møller Michael B. , Skjødt Karsten , Mortensen Leif S. , Pedersen Niels T.
Published: March 2003

The cyclin‐dependent kinase inhibitor p27Kip1 is a negative cell cycle regulator linking extracellular growth‐regulatory signals to the cell cycle machinery in G1. We investigated the pattern and prognostic value of p27Kip1 expression in a population‐based group of 203 non‐Hodgkin's lymphoma (NHL) patients. The expression of p27Kip1 was identified by immunohistochemistry and correlated with Ki‐67 expression and clinical features. Correlation with outcome was determined using uni‐ and multivariate analysis stratified by clinical grade. Except for very aggressive NHL, there was a negative correlation between p27Kip1 and Ki‐67 expression. Low expression of p27Kip1, defined as nuclear p27Kip1 expression in <40% of malignant cells, was predictive of poor survival in indolent and aggressive NHL. However, even in this regard, very aggressive lymphomas behaved differently as those with low p27Kip1 expression tended to do better. Likewise, a high proliferation rate (Ki‐67 >40%) was associated with poor survival in indolent and aggressive lymphomas. Multivariate analysis using the proportional hazards model showed that only p27Kip1, and not Ki‐67, maintained independent prognostic significance in indolent and aggressive lymphomas (relative risk = 2.0; P = 0.0095). The low cost and simplicity of this standard immunohistochemistry analysis makes p27Kip1 a promising and suitable prognostic marker in routine diagnostic laboratories in a standard diagnostic panel.
Enhanced adherence of β‐thalassaemic erythrocytes to endothelial cells
Authors: Hovav  , Goldfarb  , Artmann  , Yedgar  , Barshtein
Published: March 2003

The adherence of red blood cells (RBC) to endothelial cells (EC), shown to correlate with microvascular occlusion in sickle cell disease and malaria, is considered a major contributor to microcirculatory disorders. In the present study the adherence to EC was markedly enhanced with RBC from β‐thalassaemia major (TM) patients, and even more so with RBC from β‐thalassaemia intermedia (TI) patients (10‐fold and 25‐fold higher than normal, respectively). It is proposed that enhanced RBC/EC adherence may contribute to the microcirculatory disorders observed in thalassaemia, especially in TI patients who are particularly known to suffer from leg ulcers.
Cost‐minimization analysis of CHOP, fludarabine and rituximab for the treatment of relapsed indolent B‐cell non‐Hodgkin's lymphoma in the U.K.
Authors: Sweetenham John , Hieke Klaus , Kerrigan Matthew , Howard Paul , Smartt Pamela F. M. , McIntyre Ann‐Marie , Townshend Sarah
Published: March 2003

The optimal therapy for patients with relapsed indolent B‐cell non‐Hodgkin's lymphoma is unclear. Combination chemotherapy such as CHOP (cyclophosphamide, doxorubicin, vincristine, prednisolone) or purine analogues including fludarabine are frequently used and the anti‐CD20 monoclonal antibody rituximab has recently been licensed for use. However, no comparative studies of these therapies have been reported. Since relapsed indolent B‐cell NHL is generally regarded as incurable with current therapies, the place of each of these therapies is likely to be determined by their relative efficacy, toxicity and cost. We undertook a literature review and a retrospective analysis of patients receiving combination chemotherapy for relapsed indolent B‐cell NHL at our institution to determine the response rates and the duration of response when treated with CHOP or fludarabine. Reported response rates and median response duration for these regimens are similar, and similar to those reported in phase II studies of rituximab. A cost minimization analysis was therefore conducted.

The per patient costs for the treatment of drug‐related adverse events were £5049 for CHOP, £2953 for fludarabine and £109 for rituximab. When costs of a full course of each treatment were compared, the costs per patient for CHOP, fludarabine and rituximab were £7210 (£5975–8445), £10022 (£8917–11126) and £6080 (£5892–6267) respectively. In this preliminary analysis, rituximab appeared to have a similar efficacy rate to CHOP and fludarabine, but had significantly fewer adverse events and a lower total cost per patient. These data require confirmation in a prospective randomized study with formal assessment of cost‐effectiveness.
Prestorage and bedside leucofiltration of whole blood modulates storage‐time‐dependent suppression ofin vitroTNFα release
Authors: Mynster  , Hammer  , Nielsen
Published: March 2003

Immunosuppression after transfusion may be related to the content of leucocytes in the transfused blood. Therefore we studied the effects of prestorage and bedside leucodepletion by filtration on the suppression by whole blood ofin vitrostimulated tumour necrosis factor alpha (TNFα) release. Nine units of whole blood were leucofiltered prestorage and stored for 35 d. 27 units, 3 × 9, were stored and leucofiltered at the bedside after 7, 21 and 35 d. Supernatants were collected from all units during storage and added to a whole blood assay ofE. coli‐LPS‐stimulated TNFα release. The effects of storage were assessed and compared with supernatants collected immediately after donation as reference. TNFα release was storage time dependently suppressed to: 81%, 74% and 57% by supernatants from non‐filtered blood stored for 7, 21 and 35 d, respectively. Prestorage leucofiltration almost eliminated this effect, but we still observed a storage‐time‐dependent suppression by bedside‐leucofiltered blood to 88%, 78% and 65%, respectively. Prestorage leucofiltration appeared to reduce storage‐time‐dependent suppression ofin vitrostimulated TNFα release induced by plasma from whole blood compared with non‐filtered and bedside‐leucofiltered whole blood.
Circulating platelet–neutrophil complexes represent a subpopulation of activated neutrophils primed for adhesion, phagocytosis and intracellular killing
Authors: Peters M. J. , Dixon G. , Kotowicz K. T. , Hatch D. J. , Heyderman R. S. , Klein N. J.
Published: March 2003

Platelets play a prominent role in linking the processes of inflammation, haemostasis and thrombosis. Recent studies have shown that platelets form heterotypic aggregates with leucocytes via platelet CD62P and leucocyte β2 integrins. These interactions have been observed in vitro in blood taken from healthy volunteers and in clinical conditions in which thrombosis and inflammation are prominent.

This study investigated the properties of platelet–neutrophil complexes (PNCs) in anticoagulated whole blood. At rest, neutrophils in PNCs exhibit a significantly more activated adhesion molecule profile than free neutrophils with increased CD11b expression and activation (increased binding of the CD11b/CD18 ‘activation reporter’ monoclonal antibody 24) and decreased CD62L expression. In addition, neutrophils in PNCs phagocytosed significantly more Neisseria meningitidis and produced more toxic oxygen metabolites than free neutrophils.

Stimulation with the platelet agonist adenosine diphosphate (ADP) led to further increases in CD11b expression and activation, loss of CD62L as well as increased phagocytosis and toxic oxygen metabolite production throughout the whole neutrophil population. When these experiments were repeated with the CD62P blocking antibody G1 the effects were inhibited to a variable extent, dependent upon the parameter under investigation. These results indicate that both soluble and contact‐dependent factors contribute to platelet‐mediated neutrophil activation.

Platelet neutrophil complexes represent a large subpopulation of neutrophils with a more activated adhesion molecule profile, and a greater capacity for phagocytosis and toxic oxygen metabolite production. This study provides further support for a role for PNCs in both health and disease.
Prevalence of TT virus in plasma pools and blood products
Authors: Pisani Giulio , Cristiano Karen , Wirz Maria , Bisso Guillermo , Beneduce Francesca , Morace Graziella , Rapicetta Maria , Gentili Giuliano
Published: March 2003

A high prevalence of TT virus (TTV), a novel virus recently identified in the serum of a patient with post‐transfusion hepatitis of unknown aetiology, has been reported in blood donors worldwide. We investigated the presence of TTV DNA in several lots of blood products and in the corresponding plasma pools. In the process, we determined, from three sets of primers, the one which was most efficient in detecting the viral nucleic acid. This set amplifies the region closest to the 3′‐end of the TTV genome which was proved, by sequence analysis, to be more conserved than the other two regions. Whereas all 10 intravenous immunoglobulin and 21 albumin batches were TTV negative, 4/5 factor VIII concentrates and 4/10 intramuscular immunoglobulin batches were TTV positive. A high prevalence of TTV DNA (70%) was found in the plasma pools that were collected from four different countries. These results confirm the worldwide distribution of this virus and show that TTV is removed with a varying efficiency during the manufacture of blood products.
A combination of megakaryocyte growth and development factor and interleukin‐1 is sufficient to culture large numbers of megakaryocytic progenitors and megakaryocytes for transfusion purposes
Authors: Van Den Oudenrijn Sonja , De Haas Masja , Calafat Jero , Van Der Schoot C. Ellen , Von Dem Borne Albert E. G. KR.
Published: March 2003

Chemotherapy‐induced thrombocytopenia is a major risk factor in cancer treatment. The transfusion of autologous ex vivo expanded megakaryocytes could be a new therapy to shorten the period of thrombocytopenia. Therefore we investigated, in a liquid culture system, the effect of various cytokine combinations composed of pegylated megakaryocyte growth and development factor (PEG‐rHuMGDF), interleukin‐1 (IL‐1), IL‐3, IL‐6, IL‐11 and stem cell factor (SCF) on the proliferation and differentiation of CD34+ cells, in order to define the most optimal and minimum levels of cytokine combinations for megakaryocyte expansion.

Besides PEG‐rHuMGDF, IL‐1 was found to be important for optimal megakaryocyte expansion. Depletion of either SCF, IL‐6 or IL‐11 did not exert a large effect, but the absence of IL‐1 strongly diminished the number of megakaryocytic cells. Addition of IL‐3 to the combination PEG‐rHuMGDF, IL‐1, IL‐6, IL‐11 and SCF significantly reduced the number of megakaryocyte progenitors (CD34+CD41+ cells) and the number of CFU‐Meg. Furthermore, we found a strong correlation between the number of CD34+CD41+ cells and the number of CFU‐Meg obtained after 8 d culture.

Our study shows that optimal ex vivo expansion of megakaryocytes is achieved by the combination of PEG‐rHuMGDF and IL‐1. The numbers of megakaryocytes and megakaryocyte progenitors (CD34+CD41+) obtained in our liquid culture system with the growth factor combination PEG‐rHuMGDF and IL‐1 are suitable for transfusion purposes.
Clonal immunoglobulin light chain variable region germline gene use in AL amyloidosis: association with dominant amyloid‐related organ involvement and survival after stem cell transplantation
Authors: Comenzo Raymond L. , Wally Jeremy , Kica Geraldina , Murray Jessica , Ericsson Thomas , Skinner Martha , Zhang Yana
Published: March 2003

AL (primary or immunoglobulin light chain) amyloidosis (AL) differs from myeloma per se in that tissue deposits of amyloid are found, typically in association with small numbers of clonal plasma cells producing λ light chains, and also in that AL patients typically present with a predominantly dysfunctional organ‐system. This constellation of features — fibrillar deposits comprised of light chains, λ light chain predominance, and organ‐system tropism and dysfunction — remains unexplained. Select patients with AL respond to haemopoietic stem cell transplantation (SCT) with clinical improvement and extended survival, particularly those who do not have cardiac involvement. In order to ascertain whether the organ‐system tropism of AL was associated with immunoglublin light chain variable region (Ig VL) germline gene utilization, we attempted to clone, sequence and assign germline donors to the clonal Ig VL genes of 62 AL patients, all of whom were treated with SCT. We succeeded in 39 cases, identifying clonal AL genes derived from donors of the λI (n = 10), λII (n = 5), λIII (n = 6), λVI (n = 11) and KI (n = 7) subtypes. The majority of the donors (IGLV6S1, DPL5, DPL2,DPL23 and LFVK431) were genes that appear in the expressed repertoire <5% of the time, suggesting an intrinsic propensity to form amyloid under certain conditions. Patients whose clones derived from the λVI IGLV6S1 donor uniformly presented with dominant renal involvement while those with other Vλ or unknown donors often had dominant cardiac or other organ involvement, particularly patients whose clones derived from the λI DPL2 donor. In addition, both early (<3 months) and overall post‐SCT survival were significantly better in λVI IGLV6S1 patients compared to patients with other Vλ donors. These findings indicate that there are important associations in AL amyloidosis among Ig VL gene utilization, organ‐system tropism and post‐SCT survival.
Phenotypic and molecular analysis of six human cell lines derived from patients with plasma cell dyscrasia
Authors: Gooding R. P. , Bybee A. , Cooke F. , Little A. , Marsh S. G. E. , Coelho E. , Gupta D. , Samson D. , Apperley J. F.
Published: March 2003

Cell lines RPMI 8226, JJN3, U266 B1, NCI‐H929 (all EBV−) and ARH77 and HS‐Sultan (both EBV+) have been extensively characterized in this study. EBV− lines expressed the phenotype (CD138−, CD19+, CD20+) whereas EBV+ were (CD138+, CD19−, CD20−). CD56 expression was restricted to EBV− cell lines, with the exception of U266 B1, whereas PCA‐1 was strongly expressed on five of the six cell lines. Only EBV+ cell lines bound peanut‐agglutinin (PNA). However, all cell lines bound the lectin Jacalin that binds the same receptor as PNA, irrespective of the receptors sialylation status. By RT‐PCR and direct sequencing of their IgH V/D/J domains, ARH77 was demonstrated to use the germline sequence VH4‐34/dm1/JH6b, whereas no arrangement was demonstrated for RPMI 8226, suggesting IgH gene deletion or mutation. HLA class I and II antigens were detected using HLA typing on all cell lines warranting their use as suitable targets for HLA‐restricted cytotoxic T cells. By sensitive RT‐PCR, mRNA for IL‐6, IL‐6R and TNFβ was found expressed in all cell lines. IL‐1 mRNA expression was predominantly associated with the EBV+ phenotype. Although mRNA for IL‐3 and GM‐CSF was never detected, transcripts for c‐kit ligand and, more commonly, its receptor were. Likewise GM‐CSF, M‐CSF and erythropoietin mRNA transcripts were detected in the majority of cell lines.
Measurement of endogenous and exogenous alpha‐granular platelet proteins in patients with immune and nonimmune thrombocytopenia
Authors: Hughes Mary , Hayward Catherine P. M. , Horsewood Peter , Warkentin Theodore E. , Kelton John G.
Published: March 2003

Idiopathic thrombocytopenic purpura (ITP) is caused by antiplatelet antibodies and is characterized by increased platelet destruction and elevated levels of IgG (platelet‐associated IgG, PAIgG). Nonimmune thrombocytopenic patients also have elevated levels of PAIgG. In this study we investigated two possible biological explanations for the increased levels of PAIgG in these patients. The first hypothesis suggests that a thrombocytopenic stress causes increased thrombocytopoiesis with increased numbers and content of the platelet alpha granules. The second hypothesis is that for uncertain reasons (immunological or cytokine) there is increased absorption of plasma proteins by either megakaryocytes or by the platelets themselves. To address this issue, we compared the level of megakaryocyte synthesized alpha granular proteins [platelet factor 4 (PF4) and beta‐thromboglobulin (β‐TG)] to plasma‐absorbed alpha granular proteins (albumin, IgG and fibrinogen) in patients with immune (n = 39) and nonimmune (n = 60) thrombocytopenias. Plasma‐absorbed alpha‐granular proteins were elevated in both immune and nonimmune thrombocytopenia with no increase in megakaryocyte synthesized alpha‐granular proteins. These plasma‐derived protein elevations were not attributable to elevated mean platelet volumes or elevated plasma concentrations of the respective protein. We hypothesize that the increased IgG in these platelets is not the result of production of larger platelets, but reflects a selective increase in the endocytosis of plasma‐absorbed alpha‐granular proteins at the megakaryocyte and/or platelet level.
Secondary leukaemia and myelodysplasia after autografting for lymphoma: results from the EBMT
Authors: Milligan Donald W. , Ruiz de Elvira M. Carmen , Kolb Hans‐Jochem , Goldstone Anthony H. , Meloni Giovanna , Rohatiner Ama Z. , Colombat Phillip , Schmitz Norbert
Published: March 2003

Between 1978 and 1996 more than 7500 lymphoma transplants have been reported to the European Bone Marrow Transplantation (EBMT) Lymphoma Registry. This has been examined to establish the incidence of secondary leukaemia and myelodysplasia and to relate this to possible prognostic factors. 131 centres representing 4998 patients responded to a questionnaire. This identified 66 patients with post transplant myelodysplastic syndrome (MDS)/acute myeloid leukaemia (AML). The actuarial risk for MDS/AML at 5 years post‐transplant (±95% CI) was 4.6% (3.1–6.8) for Hodgkin's disease and 3.0% (2.0–4.3) for non‐Hodgkin's lymphoma. Multivariate analysis for all patients demonstrated an effect of age at transplant, radiotherapy at conditioning, number of transplants and interval between diagnosis and transplant as risk factors. For patients with NHL, grade of histology was important (low grade  >  intermediate or high‐grade); for Hodgkin's disease, female sex was identified as a risk factor. These findings suggest that the incidence of MDS/AML may not be greater following an autograft than after conventional chemotherapy.
Bryostatin and CD40‐ligand enhance apoptosis resistance and induce expression of cell survival genes in B‐cell chronic lymphocytic leukaemia
Authors: Kitada Shinichi , Zapata Juan M. , Andreeff Michael , Reed John C.
Published: March 2003

Modulating signal transduction pathways represents a promising approach for altering the biological behaviour of haemopoietic malignancies. B‐cell chronic lymphocytic leukaemia (B‐CLL) cells were treated in vitro with CD40‐ligand (CD40L) (CD154) or the protein kinase C modulator Bryostatin‐1, exploring the effects on: (a) sensitivity to apoptosis induction by chemotherapeutic drugs (fludarabine, dexamethasone) or anti‐Fas antibody; (b) expression of apoptosis‐regulatory proteins (Bcl‐2, Bcl‐X, Mcl‐1, Bax, Bak, BAG‐1, Flip, XIAP); (c) expression of cell surface co‐stimulatory antigens (CD80 [B7.1]; CD54 [ICAM‐1]; CD70); and (d) expression of immune modulatory receptors (CD27, CD40, CD95 [Fas]). CD40L and Bryostatin decreased both spontaneous and drug‐induced apoptosis in most B‐CLL specimens tested. Apoptosis resistance was associated with CD40L‐ and Bryostatin‐induced elevations in the anti‐apoptotic Bcl‐2 family protein Mcl‐1. CD40L also induced striking increases in the levels of the anti‐apoptotic protein Bcl‐XL in B‐CLLs. CD40L stimulated increases in the surface expression of CD40, CD54, CD69, CD70, CD80 and CD95, whereas Bryostatin induced expression of CD40, CD54, CD69 and CD95 but not the co‐stimulatory molecules CD70 and CD80. Despite elevations in the expression of CD95 (Fas), anti‐Fas antibodies failed to induce apoptosis of CD40L‐ and Bryostatin‐treated B‐CLL cells. This Fas‐resistance was associated with increased expression of the Fas‐antagonist Flip in CD40L‐treated, and with elevations in the caspase inhibitor XIAP in Bryostatin‐treated B‐CLLs. The potential anti‐apoptotic properties of CD40L and Bryostatin should be taken into consideration when employing these agents in clinical trials involving patients with B‐CLL.
WASP is involved in proliferation and differentiation of human haemopoietic progenitors in vitro
Authors: Kajiwara Michiko , Nonoyama Shigeaki , Eguchi Mitsuoki , Morio Tomohiro , ImAi Kohsuke , Okawa Hiroji , Kaneko Masafumi , Sako Masahiro , Ohga Shouichi , Maeda Miho , Hibi Shigeyoshi , Hashimito Hisako , Shibuya Atsushi , Ochs Hans D. , Nakahata Tatsutoshi , Yata Jun‐Ichi
Published: March 2003

The Wiskott‐Aldrich syndrome (WAS) is an X‐linked recessive disorder characterized by thrombocytopenia, immunodeficiency and eczema. X‐linked thrombocytopenia (XLT) is a mild form of WAS with isolated thrombocytopenia. Both phenotypes are caused by mutation of the Wiskott‐Aldrich syndrome protein (WASP) gene. In this study we investigated the role of WASP in the differentiation of CD34‐positive (CD34+) cells isolated from the bone marrow of patients with WAS (n = 5) or with XLT (n = 4). Megakaryocyte colony formation was significantly decreased in patients with WAS when compared with normal controls. The formation of granulocyte‐macrophage colonies and erythroid bursts were also decreased in WAS patinets. In contrast, in XLT patients, formation of all these colonies was normal. However, in vitro proplatelet formation of megakaryocytes induced by thrombopoietin was markedly decreased in both XLT and WAS. Electron microscopic examination revealed that megakaryocytes obtained from WAS or XLT patients grown in vitro had abnormal morphologic features, which seemed to be caused by defective actin cytoskeletal organization, including labyrinth‐like structures of the demarcation membrane system and deviated distribution of the α‐granules and demarcation membrane system. These observations indicate that WASP is involved in the proliferation and differentiation of CD34+ haemopoietic progenitor cells probably by its participation in signal transduction and in the regulation of the cytoskeleton.