bluuebunny/crossref_metadata_2025 huggingface dataset:
T cell function in children with acute lymphoblastic leukaemia
Authors: Nash K. A. , Mohammed G. , Nandapalan N. , Kernahan J. , Scott R. , Craft A. W. , Toms G. L.
Published: March 2008

Summary. Modern intensive chemotherapy has dramatically improved the prognosis of acute lymphoblastic leukaemia (ALL) in children. However, once remission has been established, quality of life and even survival may be threatened by exacerbation of viral infections in the prolonged period of continuation therapy necessary to prevent relapse. Often the viruses involved in the most severe infections are from the herpesvirus and paramyxovirus groups, suggesting that patients suffer from a defect in the cellular immunity thought essential to control such cell‐associated infections. This may result from a T cell defect and, in this study, T cell responsiveness of patients under therapy for leukaemia has been investigated.

In vitro proliferative responses of peripheral blood leucocytes (PBL) to the T cell mitogen phytohaemagglutinin (PHA) were impaired in children with ALL before treatment and in the induction of remission. Impairment was attributable to reduced T cell numbers, the presence of inhibitors in the patient's serum and direct damage to lymphocytes. On achieving remission, proliferative responses to PHA of both CD4+ and CD8+ T cell subsets quickly returned to normal levels with the switch to continuation chemotherapy. Proliferative responses to Herpes simplex virus antigens were also apparently normal in the majority of patients tested in remission.

Further investigations, however, have suggested a persisting defect in CD8+ lymphocyte function. Gamma interferon secretion by PHA‐stimulated PBLs was severely reduced for children with ALL in remission when compared with control children of similar age. Further, cytotoxic T lymphocyte responses to allogeneic cells could only be induced in PBL isolated from two of 13 children in remission from ALL whilst all control children of similar age and adults produced anti‐allogenetic responses.
High IL‐6 levels in ascitic fluid correlate with reactive thrombocytosis in patients with epithelial ovarian cancer
Authors: Gastl G. , Plante M. , Finstad C. L. , Wong G. Y. , Federici M. G. , Bander N. H. , Rubin S. C.
Published: March 2008

Summary. Non‐haematopoietic malignancies are commonly associated with thrombocytosis. The aetiology of tumour‐associated thrombocytosis is still unclear but may be related to tumour‐derived thrombopoietin‐like factors. Epithelial ovarian tumour cells have been shown to release IL‐6 in vitro and high IL‐6 levels have been identified in ascites of patients with ovarian cancer. Since IL‐6 is a potent stimulator of megakaryocytopoiesis we examined IL‐6 production at the tumour site and its relationship to serum IL‐6 levels and circulating platelet counts in patients with ovarian cancer. Forty patients undergoing exploratory laparotomy for epithelial ovarian cancer [stage I + II: 6 (15%); stage III: 25 (62.5%); stage IV: 9 (22.5%)] and 24 women with benign ovarian conditions were evaluated. Sera were available from 39 cases with ovarian cancer and from 19 cases with benign ovarian tumours. Ascites was obtained from 35 patients with ovarian cancer. IL‐6 activity in serum and ascitic fluid was determined by the standard B9 proliferation assay (detection level: 1 pg/ml). IL‐6 bioactivity was detectable in 22 (56%) sera from patients with ovarian cancer, but in only five (26%) of the serum samples obtained from benign cases (P < 0.001). Serum IL‐6 levels in patients with ovarian cancer were significantly higher (median 3 pg/ml; range < 1 to 1221 pg/ml) than in patients with benign ovarian conditions (median 0 pg/ml; range < 1 to 4 pg/ml) (P < 0.001). However, much higher concentrations of IL‐6 were measured in malignant ascites specimens (median 22 100 pg/ml; range < 1 to 182 600 pg/ml). IL‐6 bioactivity in serum and ascites samples was completely inhibited by a neutralizing goat antihuman IL‐6 antiserum. Thrombocytosis (platelet counts > 400 × 109/l) occurred in 25 (62.5%) of the 40 patients with ovarian cancer, but in only two (8%) of the 24 cases with benign ovarian tumours. In eight (20%) cases with malignant disease platelet counts ranged between 600 × 109/l and 1060 × 109/l. IL‐6 bioactivity in ascitic fluid correlated significantly with circulating platelet counts (r = 0.5916; P < 0.001). Maximum IL‐6 bioactivity in ascites and highest platelet counts occurred in patients with undifferentiated ovarian adenocarcinoma or advanced disease. In conclusion, these observations strongly suggest a role for IL‐6 in the development of tumour‐associated thrombocytosis.
Sickle cell disorders and chronic intravascular haemolysis are associated with low plasma heparin cofactor II
Authors: Porter J. B. , Young L. , Mackie I. J. , Marshall L. , Machin S. J.
Published: March 2008

Thrombotic events are known to be increased in patients with sickle cell syndromes and a variety of abnormalities of coagulation or endothelial function have been described, although the relevance of these findings either to the pathogenesis of vaso‐occlusive phenomena or the risk of thrombosis are unclear. Heparin cofactor II (HCII) and antithrombin III are circulating inhibitors of thrombin and low plasma levels have been associated with an increased risk of thrombosis in otherwise healthy individuals. We describe for the first time abnormally low plasma levels of HCII in patients with sickle cell syndromes. 45 adult patients with sickle cell syndromes (31 SS, 10 SC, 4 SβThal) were compared with 61 age matched control patients for HCII in plasma. There was a highly significant reduction in HCII in SS patients irrespective of crisis or transfusion state 0.68 ± 0.15 U/ml (mean ± SD) compared with controls 1.00 ± 0.19 U/ml (P < 0.001). HCII antigen was also significantly reduced (0.53 ± 0.19 U/ml) compared with controls (1.02 ± 0.23 U/ml, P < 0.0001). By contrast there was no reduction in antithrombin III in this group. HCII (0.63 ± 0.13 U/ml, P < 0.001) and HCII antigen (0.54 ± 0.08 U/ml, P < 0.001) are also significantly reduced in SC patients HCII levels increased towards control values during sickle cell crises, in patients taking the contraceptive pill, or with regular blood transfusion; however, plasma HCII concentrations were not increased acutely by exchange transfusion. HCII was also decreased in thalassaemia intermedia and pyruvate kinase deficiency, suggesting that intravascular haemolysis may be the cause of reduced HCII levels.
Transient and persistent expansions of large granular lymphocytes (LGL) and NK‐associated (NKa) cells: the Yorkshire Leukaemia Group study
Authors: Scott Colin Stephen , Richards Stephen J. , Sivakumaran M. , Short M. , Child J. A. , Hunt Kay M. , McEvoy M. , Steed Andrew J. , Balfour I. C. , Parapia L. A. , McVerry B. A. , Bynoe A. G. , Galvin M. C. , Norfolk Derek R. , Roberts Bryon E.
Published: March 2008

Summary. A survey of 870 different adult blood samples (primarily from patients with non‐haematological disorders) found that 269 (31%) had increased proportions (>25%) and/or absolute numbers (>1.0 × 109/l) of morphologically‐defined large granular lymphocytes (LGL), and/or pheno‐typically‐defined NK‐associated (NKa) cells. Of these, 112 were re‐analysed at least 6 months after initial presentation and were classified as‘persistent’(92/112) or ‘transient’(20/112) according to whether or not the original abnormality was still present. Lymphocyte counts in most patients with persistent abnormalities were within normal limits (18/92) or slightly increased (68/92), with only six having a lymphocytosis exceeding 10.0 × 109/l. With the exception of five persistent LGL expansions in which the granular lymphocytes did not express NKa determinants (designated LGL+NKa‐), the remaining 87 cases could be phenotypically grouped according to their primary abnormality as CD8+NKa+ (n = 33), CD4+ NKa+ (n=14), CD8dim+NKa+ (n=7) or CD8−NKa+ (n=33). TCR genotypic studies in 58 patients showed that the 16 patients with rearranged TCR components were restricted to the CD8+NKa+ group and that, in most of these, the CD8+ fraction showed abnormal relative CD16/CD56 expression. Persistent neutropenia (n=15) also appeared to be associated with primary abnormalities of CD8+NKa+ cells (12/15). with 10 of these additionally showing rearranged TCR genes. In contrast, persistently increased CD8dim+NKa+ and CD8–NKa+ components did not appear to phenotypically differ from their corresponding ‘counterparts’in normal bloods or in patients with transient LGL/NKa+ abnormalities. This survey has therefore established that persistent LGL/NKa+ abnormalities are considerably more common than suggested in published work, that a high proportion of patients with expanded CD8+NKa+ components, with quite diverse clinical histories, show evidence of clonal lymphoid populations, and that the clonal nature of such disorders appears to be associated with abnormal NKa phenotypic patterns.
Blood film features of sickle cell–haemoglobin C disease
Authors: Bain Barbara J.
Published: March 2008

Summary. The blood films of patients with sickle cell‐haemoglobin C disease (SC) were compared with those of patients with related haemoglobinopathies in order to establish the most characteristic features. The blood films from SC patients generally permitted the distinction from sickle cell anaemia (SS) but not necessarily from haemoglobin C disease (CC) or C/β thalassaemia. About half of the SC films showed characteristic SC poikilocytes and a minority also had some cells containing haemoglobin C crystals. In addition, SC differed from SS in having fewer classical sickle cells and nucleated red cells, more irregularly‐contracted cells and less polychromasia and evidence of hyposplenism.
CD2+ CD19+ acute lymphoblastic leukaemia in 16 children and adults: clinical and biological features. The Groupe d'Etude Immunologique des Leucémies (G.E.I.L.)
Authors: Lenormand B. , Vannier J. P. , Bene M. C. , Falkenrodt A. , Bayle C. , Favre M. , Garand R. , Griveau A. M. , Briquet M. T. , Reman O. , Boutard P. , Tilly H. , Faure G. , Monconduit M.
Published: March 2008

A small population of CD2+CD19+ lymphoid cells have been suggested to be common lymphoid progenitors. CD2+CD19+ biphenotypic ALL account for less than 2% of ALL. We analysed the clinical and laboratory features of a series of 16 patients with CD2+CD19+ ALL.

The incidence of tumoural syndrome was comparable to a previously published series of pre B‐ALL but significantly different from that of T‐ALL. The mean age of the 11 children of this series was 101 ±46 months, and differed significantly from that of children with pre B‐ALL (P<0.01). Complete remission was obtained for all patients except two adults. Only three relapses have been observed.

Regardless of the presence of CD2+, the 16 ALL could be classified as pre B‐ALL, according to the nomenclature used by the GEIL. Nine samples could be analysed by Southern blotting. Seven had rearranged IGH genes, usually on both chromosomes. IGK rearrangement was observed in three cases. Only one case had rearranged both TCRG and TCRβ. The patterns observed here and those reported previously follow that of the pre B‐ALL which confirms the engagement of most CD2+CD19+ biphenotypic ALL in the B‐lineage.
Ultraviolet‐B irradiation of platelets induces a dose‐dependent increase in the expression of platelet activation markers with storage
Authors: Grijzenhout M. A. , Aarts‐Riemens M. I. , Akkerman J. W. N. , Nieuwenhuis H. K. , Weelden H. van , Prooijen H. C. van
Published: March 2008

Summary. Ultraviolet B (UVB) irradiation of platelet concentrates (PCs) has been proposed as a novel technology to prevent HLA sensitization. We have recently reported that 2 J/cm2 of UVB radiation abolishes alloreactive lymphocyte responses in vitro. In order to increase the efficacy of UV irradiation for the prevention of HLA sensitization, we exposed PCs to 4 or 8 J/cm2 of UVB and evaluated the effect of UV radiation on platelet integrity during storage. We report here that UV exposed platelets show a progressive increase in the expression of activation markers P‐selectin (GMP‐140; CD62) and LIMP‐CD63 (GP‐53; CD63) on the platelet membrane over time in a dose‐dependent manner compared to age‐matched controls. Platelet metabolism was also enhanced as evidenced by significant changes in lactate and pH during post‐irradiation storage. Based on these findings we transfused PCs within 4 h after UV irradiation. PCs exposed to 4 J/cm2 showed normal post‐transfusion recoveries and haemostatic functions, while poor platelet recoveries were found after administration of PCs exposed to 8 J/cm2. We hypothesize that the rapid expression of P‐selectin on platelets exposed to the higher dose of UVB leads to an increased binding of these platelets to leucocytes in the circulation resulting in poor platelet recoveries.
Decay‐accelerating factor and CD59 expression in peripheral blood cells in aplastic anaemia and report of a case of paroxysmal nocturnal haemoglobinuria secondary to aplastic anaemia
Authors: Fujioka Seitoku , Yamada Teruo
Published: March 2008

Summary. Using flow cytometry, decay‐accelerating factor (DAF) and CD59 have been measured in peripheral blood cells of 11 patients with aplastic anaemia and 34 healthy controls. Ten of the patients had monophasic fluorescence profiles similar to those of the controls. However, one patient had small DAF‐ and CD59‐negative populations in granulocytes and monocytes, but not in erythrocytes. One year after the first assay, a second flow cytometric study revealed that all peripheral blood cell species, including erythrocytes, contained DAF‐ and CD59‐deficient populations. At this time. a sucrose haemolysis test was positive. This is the first reported case of aplastic anaemia‐PNH syndrome in which DAF‐ and CD59‐negative cells appeared first in granulocytes and monocytes and later in all types of peripheral blood cells.
Transcriptional and post‐transcriptional regulation of IL‐1β, IL‐6 and TNF‐α genes in chronic lymphocytic leukaemia
Authors: Rambaldi Alessandro , Bettoni Stefania , Rossi Vincenzo , Tini Maria‐Laura , Giudici Giovanni , Rizzo Virginia , Bassan Renato , Mantovani Alberto , Barbui Tiziano , Biondi Andrea
Published: March 2008

The present study was designed to define the mechanisms of interleukin‐1β (IL‐1β), interleukin‐6 (IL‐6) and tumour necrosis factor (TNF‐α) gene regulation in chronic lymphocytic leukaemia of B cell origin (B‐CLL). By nuclear run‐on analysis, all B‐CLL cases displayed high levels of nuclear transcription of the IL‐6 and TNF‐α genes, whereas IL‐1β gene transcription was only barely detectable. Upon in vitro culture for 1 h, B‐CLL cells from different patients were substantially heterogeneous in terms of expression of steady state mRNA levels of IL‐1β, IL‐6 and TNF‐α even though the pattern of nuclear transcription of these cytokines was only marginally affected by in vitro culture. mRNA stability was then examined and cytokine gene transcripts showed a half life of more than 2 h in cultured B‐CLL cells and treatment with cycloheximide (CHX) did not affect cytokine transcript levels in B‐CLL cells. These results indicate that: steady state levels of each mRNA do not reflect the rate of nuclear transcription of these cytokines in fresh or cultured B‐CLL cells, that purification and in vitro culture of leukaemic cells may amplify cytokine gene expression in B‐CLL, and that cytokine gene transcripts are relatively stable in B‐CLL.
CD3 down‐regulating factor in sera and culture supernatants of leukaemic cells from patients with adult T cell leukaemia
Authors: Matsuda Mitsuhiro , Maeda Yasuhiro , Shirakawa Chikashi , Masaki Hideyuki , Koyama Atsuko , Horiuchi Fusanari , Hamazaki Hiroyuki , Fujimoto Takuya , Irimajiri Kiyohiro , Horiuchi Atsushi
Published: March 2008

Immunological abnormality of T lymphocytes in patients with adult T cell leukaemia (ATL) is characterized by abnormal expression of the 55 kD chain of the receptor for interleukin 2 (IL‐2R/p55) (Tac), and the down‐regulation of CD 3 expression. Using serum and culture supernatants of leukaemic cells from ATL patients (Group A) whose CD 3 expression was down‐regulated and those (Group B) whose CD 3 was not low, the possible mechanism of CD 3 down‐regulation on ATL cells was discussed. When PBMC from normal individuals were cultured with sera from ATL patients for 24 h, CD 3 expression revealed by mean fluorescent intensity (MFI) was down‐regulated by sera from ATL patients in Group A (MFI: Pt 1 = 51.6 ± 4.5, Pt 2 = 48.0 ± 6.9, control = 96.5 ± 6.6), not by sera from patients in Group B (MFI: Pt 3 = 105.5 ± 7.9, Pt 4 = 102.5 ± 8.3, control = 96.5 ± 6.6). When normal PBMC were cultured with supernatants of leukaemic cells from ATL patients in Group A, this CD 3 down‐regulating activity was also detected (MFI: Pt 1 = 78.0 ± 10.2, Pt 2 = 70.6 ± 8.7, control = 94.0 ± 6.6). By using gel‐chromatography, the fractionated supernatants from ATL patients in Group A decreased CD 3 expression of normal PBMC significantly (MFI: Pt 1 = 22.9 ± 5.8, Pt 2 = 28.8 ± 7.4, control = 92.1 ± 9.6). This CD 3 down‐regulating activity in fractionated supernatant was not inhibited by any lymphokine antibodies, anti‐IL‐1α antibody (Ab), anti‐IL‐1B Ab, anti‐IL‐2 Ab, anti‐IL‐3 Ab, anti‐IL‐4 Ab, anti‐IL‐6 Ab, anti‐TNF‐α Ab and anti‐IFN‐γ Ab. Any known cytokines (IL‐1, IL‐2, IL‐3, IL‐4, IL‐6, TNF‐α and IFN‐γ) could not modulate CD 3 expression of normal PBMC. These findings suggested that there are novel factor(s) with CD 3 down‐regulating activity in the serum and culture supernatant of ATL patient and those factor(s) are involved in progression of ATL.
C‐fms protein expression by B‐cells, with particular reference to the hairy cells of hairy‐cell leukaemia
Authors: Till K. J. , Lopez A. , Slupsky J. , Cawley J. C.
Published: March 2008

Although the hairy cells (HCs) of hairy cell leukaemia (HCL) are now thought to be a form of activated B cell, they have long been known to possess certain monocytoid characteristics. Since the proto‐oncogene c‐fms is a feature of cells of the monocyte/macrophage lineage, we examined HCs for c‐fms expression. We found that approximately 80% of peripheral blood HCs expressed the c‐fms protein (8/8 cases). Expression of the 150 kD protein by HCs was shown using three different techniques, APAAP, immunofluorescence and immunoprecipitation, using two different antibodies. Other mature B cell lymphoproliferative disorders examined (PLL, CLL and multiple myeloma) did not express c‐fms. We also examined the c‐fms expression of normal B‐cells: both the in vivo activated (low density) fraction of tonsil B cells and tonsil B cells activated in vitro with SAC plus IL‐2 expressed the c‐fms protein. As in the case of monocytes c‐fms expression by HCs was shown to be down regulated by its ligand M‐CSF, and by TNFα, both caused a decrease in the receptor expression from 80% to 30% and in the intensity of staining from 6 to 3 x 104 molecules/cell. However, as for monocytes, GM‐CSF treatment of HCs had no effect on the expression of c‐fms: αIFN also had no effect. M‐CSF treatment of HCs also induced phosphorylation of c‐fms, and a number of other proteins, on tyrosine. However, M‐CSF was unable to induce HC proliferation either alone or in combination with IL‐2, IL‐4 or IL‐6; in addition it had no effect on HC proliferation induced by SAC, anti‐μ or TNFα. In addition, M‐CSF either alone, or in combination with the above cytokines, had no effect on the differentiated state of HCs as shown by both immunoglobulin secretion and surface antigen expression. M‐CSF also had no effect on the morphology or long‐term survival of HCs in culture. This study therefore demonstrates that both HCs and activated B‐cells express c‐fms, and that M‐CSF and binds to and activates its receptor on HCs. Although c‐fms and several other proteins were shown to be phosphorylated in response to M‐CSF, the functional consequences of this phosphorylation remain unclear.
Immunophenotyping of low‐grade B‐cell lymphoma in blood and bone marrow: poor correlation between immunophenotype and cytological/histological classification
Authors: Schwonzen M. , Pohl C. , Steinmetz T. , Seckler W. , Vetten B. , Thiele J. , Wickramanayake D. , Diehl V.
Published: March 2008

Results of immunophenotypic examinations of peripheral blood and/or bone marrow (BM), involved in low‐grade B‐cell non‐Hodgkin's lymphomas, were compared with the results of cytomorphological and histopathological examinations in 133 adult patients. 69 cases of chronic B‐lymphocytic leukaemia (B‐CLL), 16 centrocytic (CC) lymphomas, 14 centroblastic‐centrocytic (CB/CC) lymphomas, 15 immunocytomas (IC), 10 cases of hairy cell leukaemia (HCL), four prolymphocytic leukaemias (PLL), two B‐CLL in transformation, one splenic lymphoma with villous lymphocytes (SLVL), one hairy cell leukaemia variant (HCL‐V), and one lymphocytic lymphoma (LC) were classified according to the Kiel and /or FAB classification. Leukaemic disease was found in 105 cases. The following markers were used for immunocytology (APAAP technique) of blood and/or BM smears: CD 19, CD 5, CD 10, CD 11c, CD 14, CD 21, CD 22, CD 23, CD 25, CD 38 and TdT. All cases tested showed CD 19, but no TdT expression. Every case of HCL had a distinct phenotype with expression of CD 11c, CD 22 and CD 25 and the lack of CD 5 and CD 23 antigens. In all other NHL cases a very heterogenous expression of CD‐antigens with no significant correlations to the cytomorphological subtypes was found. The expression of CD 5 is a frequent but inconstant finding in lymphoproliferative diseases other than B‐CLL, so 50% of CB/CC, 75% of CC and 80% of IC were CD 5 positive. Our results indicate that, with the exception of HCL, the diagnostic relevance of immunophenotyping for the classification of cytomorphologically and histopathologically defined subtypes in blood and/or BM is of very limited value.
Cyclophosphamide and etoposide therapy with GM‐CSF for VAD‐resistant multiple myeloma
Authors: Dimopoulos Meletios A. , Delasalle Kay B. , Champlin Richard , Alexanian Raymond
Published: March 2008

Few effective regimens are available for patients with advanced multiple myeloma resistant to or relapsing after both alkylating agents and VAD. We treated 52 patients with advanced and refractory multiple myeloma with the combination of cyclophosphamide (3.0 g/m2) and etoposide (900 mg/m2) followed by GM‐CSF at a daily dose of 0.125 mg/m2 until recovery of granulocytes. 42% of patients responded with a median time of 19 d for recovery of granulocytes to 0.5 x 109/1 and a 4% mortality rate. Eight responding patients received a second myeloablative treatment supported by either autologous bone marrow (six patients) or blood stem cells (two patients). The median survival time for all patients was 11 months and the median remission time for responding patients was 8 months. The combination of cyclophosphamide and etoposide provided an effective rescue treatment for many patients with advanced multiple myeloma resistant to conventional therapies. This programme also allowed early marrow or blood stem cell collection in support of subsequent myeloablative therapy for selected patients.
Cisplatin‐induced platelet activation requires mononuclear cells: role of GMP‐140 and modulation of procoagulant activity
Authors: Yen Tina , Walsh John D. , Pejler Gunnar , Berndt Michael C. , Geczy Carolyn L.
Published: March 2008

Cytotoxic drugs may potentiate the thrombotic complications in patients with malignancies and platelet function abnormalities have been reported after initiation of cisplatin therapy. This report describes a prolonged activation of platelets over 6‐24 h co‐culture with peripheral blood mononuclear cells (PBM) by pharmacological doses of cisplatin. Cisplatin had no direct effect on platelets and depended on PBM to produce aggregation which was apparently not mediated by products of the cyclooxygenase or lipoxygenase pathways, by platelet activation factor (PAF) or by thrombin. Although platelet aggregation normally involves the binding of fibrinogen to the β3 integrin, GP IIb‐IIIa, on activated platelets, the cisplatin‐dependent platelet aggregation observed in the co‐culture experiments was not inhibited by anti‐GP IIb‐IIIa monoclonal antibody which blocks fibrinogen‐dependent aggregation nor by an adhesive peptide containing the RGDS integrin recognition sequence. Rather, aggregation appeared to involve a novel 140 kD granule membrane protein (GMP‐140) mediated mechanism since aggregation was almost completely blocked by Fab fragments of antibody to GMP‐140 and was inhibited by fluid‐phase GMP‐140. At concentrations of cisplatin, adriamycin, and LPS that induced equivalent levels of tissue factor of blood monocytes, prothrombinase activity was significantly greater in cultures containing cisplatin. Prothrombinase activity was dependent on the presence of platelets and the rate of thrombin formation was enhanced by factor Xa generated by the tissue factor‐factor VIIa complex. These studies suggest that the vascular and thrombotic complications associated with cisplatin therapy are mediated, at least in part, by platelet activation and aggregation and monocyte procoagulant activity.
The fibrinolytic response to ancrod therapy: characterization of fibrinogen and fibrin degradation products
Authors: Prentice C. R. M. , Hampton K. K. , Grant P. J. , Nelson S. R. , Nieuwenhuizen W. , Gaffney P. J.
Published: March 2008

Ancrod is a purified coagulant venom which renders blood incoagulable by cleaving fibrinopeptide A (FPA) from fibrinogen, but the mechanism involved in the clearance of fibrin from the circulation is unknown. To investigate the fibrinolytic response to ancrod, and to increase understanding of clearance mechanisms, six patients with peripheral vascular disease causing claudication were infused with ancrod at 2 u/kg over 6 h followed by 2 u/kg at 12 h intervals for 38 h. Venous blood samples were taken at time 0, 3, 6, 25 and 49 h for assay of fibrinogen (Fbg), fibrinopeptide A (FPA), total fibrin(ogen) degradation products (TDP), fibrin degradation products (FbDP), fibrinogen degradation products (FgDP), cross‐linked fibrin degradation products (XL‐FDP), tissue plasminogen activator (tPA), urinary type plasminogen activator (u‐PA), plasminogen, α2 antiplasmin (α2AP) and plasminogen activator inhibitor‐1 (PAI‐1). Fibrinogen (median and range) was 2.3 (1.4–3.90) g/l at time 0 and thereafter was undetectable. FPA rose from 2.5 (1.8–3.6) to 600 and 188 pmol/l at 3 h and 6 h and remained elevated. TDP, FbDP and FgDP increased greatly following ancrod while there was no evidence of XL‐FDP. The surprising increase in FgDP during defibrination suggests either that fibrinogen is digested following its incorporation into circulating fibrin protofibrils or that some of the fibrin subunits in the photofibril retain one of the two fibrinopeptide A's. tPA and uPA remained unchanged. Plasminogen fell from 125 (100–155)% to 79 (40–118)% at 49 h and α2AP fell from 91 (75–107)% to 24 (10–35)% at 49 h. The level of PAI‐1 was depressed during defibrination, with the exception of the 6 h data. The results demonstrate that ancrod removes FPA from fibrinogen to produce non‐cross‐linked (soluble) fibrin. This is cleared from the circulation without evidence of an increase in the circulating activities of the plasminogen activators, tPA or UK, but with evidence of plasminogen activation and consumption.
A new platelet alloantigen, Tua, on glycoprotein IIIa associated with neonatal alloimmune thrombocytopenia in two families
Authors: Kekomäki R. , Jouhikainen T. , Ollikainen J. , Westman P. , Laes M.
Published: March 2008

We describe immunization of two mothers against a new platelet alloantigen, designated Tua, in association with thrombocytopenia in their first born children. The platelet‐specific antibodies were identified by a glycoprotein‐specific platelet protein assay with husband's platelets. Monoclonal antibodies against glycoprotein complex IIb/IIIa (AP2) and against glycoprotein IIb (SZ22) could be used to immobilize the antigen bearing protein. When monoclonal antibodies against glycoprotein Ib/IX (FMC25) or Ia/IIa (Gi9) were used, no platelet‐specific antibodies were detectable. The previously described alloantigens on the glycoprotein IIb/IIIa complex (HPA 1,3,4, Sra and Vaa) were not responsible for the reaction. Immunochemical analysis by an immunoblot assay showed that the Tua antigen resides on GPIIIa but the antigen was destroyed by reduction of the protein. Altogether 10 individuals belonging to three unrelated families were shown to carry the antigen. The family studies within three generations indicated autosomal codominant inheritance. Thus the Tua antigen is apparently different from all previously published platelet alloantigens. One Tua positive blood donor was identified in a population study of approximately 150 individuals. This indicates a low frequency in the Finnish population. Extended population studies will be required to determine a more exact frequency of Tua antigen.
Microenvironmentally dependent effects on murine haemopoiesis by a prolonged interleukin‐1 treatment
Authors: Haan G. de , Dontje B. , Loeffler M. , Nijhof W.
Published: March 2008

Summary. We administered recombinant human IL‐lβ (400 ng/d, s.c.) for 10 d to normal C57B1 mice and determined daily granuloid and erythroid parameters in marrow, spleen and blood.

In the marrow CFU‐GM numbers were not affected but later granuloid cell stages were moderately enhanced (170%). In the spleen, however, CFU‐GM numbers were sharply increased (1600%). whereas the granuloid precursors only doubled. Blood granulocytes increased transiently to 275% on day 5. In the marrow all erythroid parameters were severely reduced. This reduction was partially compensated by the spleen where initially only BFU‐E and with some delay also more mature erythroid cells accumulated. At the end of the treatment mice were slightly anaemic. When mice were treated with IL‐1 and erythropoietin (10 U/d) simultaneously, the inhibitory effects on erythropoiesis were less severe. In agreement with in vivo results, IL‐1 inhibited in vitro colony growth of CFU‐E from normal bone marrow and spleen but spleen CFU‐E from 5 d IL‐1 treated mice were insensitive.

We conclude that IL‐1 can induce stimulation or inhibition of haemopoietic progenitor cells depending on their micro‐environment.
Partial engraftment of donor bone marrow cells associated with long‐term remission of haemophagocytic lymphohistiocytosis
Authors: Landman‐Parker J. , Deist F. Le , Blaise A. , Brison O. , Fischer A.
Published: March 2008

Summary. We used polymerase chain reaction amplification of minisatellite sequences or of a Y chromosome‐specific sequence and Southern blotting to analyse long‐term engraftment (12–82 months) after bone marrow transplantation (BMT) for familial haemophagocytic lymphohistiocytosis (FHL). Six children aged from 1 to 18 months were transplanted with bone marrow from an HLA‐identical sibling in five cases and from an HLA‐nonidentical related donor (one mismatched HLA antigen) in one. The conditioning regiment included VP 16–213 (900 mg/m2), busulfan(16 mg/kg), cyclophosphamide (200 mg/kg) and, in one case. aracytine (2 g/m2).

Four patients are alive without therapy more than 3 years after BMT; the other two relapsed 1 year after BMT. DNA was extracted from separated polymorphonuclear cells and mononuclear cells, as well as from separated E + and E — cells in one case and CD16 + (natural killer) and CD16 — cells in two cases. Engraftment was partial in the four long‐term survivors. Recipient cells were largely predominant in three of them as well as in one of the patients who relapsed (the donor also developed FHL 18 months after BMT). E +, E‐, CD16 + and CD16— cells presented the same pattern of chimaerism. Engraftment failed to occur in the patient who received an HLA‐nonidentical bone marrow. These results indicate that partial engraftment is compatible with longterm remission of FHL and that the presence of a small proportion of cells of donor origin can prevent FHL‐related lymphocyte and macrophage activation.
Chronic neutropenia of childhood: frequent association with parvovirus infection and correlations with bone marrow culture studies
Authors: McClain Kenneth , Estrov Zeev , Chen Hong , Mahoney Donald H.
Published: March 2008

Summary. Children with neutropenia of more than 3 months duration often have evidence of immune‐mediated destruction of mature neutrophils and variable abnormalities of myeloid precursors in their bone marrow. These patients often have anti‐neutrophil antibodies which persist for several months. To further investigate the aetiology of neutropenia in such patients, bone marrow cells were evaluated for the presence of common viruses. Fifteen of 19 patients tested had evidence for parvovirus infection by PCR amplification of bone marrow DNA with parvovirus specific primers. Of these 15, six also had serologic evidence of parvovirus infection. Anti‐neutrophil antibodies were identified in nine of 12 patients with parvovirus infection. Bone marrow culture studies done on six patients revealed varying degrees of myeloid and erythroid inhibition by patient plasma. These studies indicate that parvovirus may be a common cause of immune‐mediated neutropenia in children.
Serum stem cell factor concentration in patients with myelodysplastic syndromes
Authors: Bowen David , Yancik Sue , Bennett Larry , Culligan Dominic , Reasser Kenneth
Published: March 2008

Summary. Stem cell factor (SCF) is characterized by its capacity to synergize dramatically with other haemopoietic growth factors in in vitro erythroid, myeloid, and lymphoid progenitor culture systems. We have measured serum SCF concentrations by enzyme immunoassay in 85 patients with myelodysplasia (MDS). Serum samples were taken in 1988–89 and in 1991–92 and stored at ‐20°C. Mean serum SCF concentration in the MDS patients was 2.81 ng/ml (range 0.6–8.0). This was significantly lower (P=0.0001) than the values for 234 normal subjects: mean 3.30 ng/ml (range 1.3–8. 0). No significant relationship between SCF concentration and peripheral blood counts, bone marrow parameters, red cell transfusion status, survival or FAB subtype was found, although a trend of decreasing SCF concentration from refractory anaemia through sideroblastic anaemia and chronic myelomonocytic leukaemia to refractory anaemia with excess blasts was seen. The reduced SCF serum concentration in some patients with myelodysplasia suggests a rationale for therapy with recombinant SCF in these patients.
Recombinant interferon alfa 2 a in the treatment of patients with early stage B chronic lymphocytic leukaemia
Authors: McSweeney E. N. , Giles F. J. , Worman C. P. , Jewel A. P. , Tsakona C. P. , Hoffbrand A. V. , Mehta A. B. , Newland A. C. , Cawley J. C. , Galvani D. , Singer C. R. J. , Grant I. R. , Norfolk D. R. , Goldstone A. H.
Published: March 2008

Summary. 18 patients with early stage, previously untreated H‐CLL were given interferon alfa (IFNα) 2a, 3 MU thrice weekly, subcutaneously. The peripheral lymphocyte count decreased in all patients. Response was delayed in three patients until they had received a median of 5 months therapy, one of whom had an initial transient increase in lymphocytes. Two patients normalized their blood lymphocyte counts, but neither achieved complete remission (CR). Responses were transient in eight patients lasting a median of 5 months (3–21). Binding anti‐IFNa antibodies were present in 9/17 patients tested (53%). Low titre binding antibodies (< 533 IBU/ml) were not associated with LHR, but high titre antibodies (>4401 IBU/ml) were. Two of 12 patients assessed had a > 3 g/1 increase in baseline serum IgG levels during IFNα therapy, one of whom reverted to pretreatment levels in association with LHR. Haematological toxicity was moderate, other than in two patients, one of whom developed autoimmune haemolytic anaemia and the other thrombo‐cytopenia. We conclude that IFNα lowers the lymphocyte count in early stage CLL, that the response may be delayed and that anti‐IFNcc antibodies may play a role in a proportion of those in whom the response is transient.
Impaired neutrophil chemotaxis in patients with thalassaemia major
Authors: Matzner Yaacov , Goldfarb Ada , Abrahamov Ayala , Drexler Ruth , Friedberg Aviva , Rachmilewitz Eliezer A.
Published: March 2008

Summary. Random and directed migration, O‐2 production. degradation and adhesion were studied in neutrophils obtained from patients with homozygous β‐thalassaemia and iron overload, in the presence or absence of thalassaemic serum. The only significant defect found was an impairment in directed chemotaxis, further depressed after addition of thalassaemic serum. The chemotactic defect was encountered in all the patients that have suffered from pyogenic infections except one, and was not correlated with the severity of the iron overload. It is suggested that the described neutrophil migration impairment may contribute to the tendency towards infection in certain patients with homozygous β‐thalassaemia.
Very early analysis of graft establishment after allogeneic bone marrow transplantation using the polymerase chain reaction
Authors: O'Keilly J. , Meyer B. , Stoner M. , Erber W. , Herrmann R. , Davies J.
Published: March 2008

Summary.We have used minisatellite polymorphisms flanking the apolipoprotein B locus and PCR to demonstrate the emergence of donor specific alleles in peripheral blood at 3–7 d post allogeneic bone marrow transplantation (BMT). This technique affords a very early indication of the establishment of stable engraftment, and may identify patients at risk of graft failure or leukaemic relapse. Patterns of T‐cell chimaerism in the first 7–14 d following transplantation may be closely associated with the development of graft‐versus‐host disease (GVHD), the graft versus leukaemia (GVL) effect and graft rejection and future application of this technique to the investigation of early T‐cell chimaerism may give further insights into these immunologicaly mediated events.
An interstitial deletion in the rearranged T‐cell receptor γ chain locus in a case of T‐cell acute lymphoblastic leukaemia
Authors: Taylor J. J. , Rowe D. , Reid M. M. , Middleton P. G.
Published: March 2008

Summary. This report describes the cloning and sequencing of the breakpoint of a deletion of approximately 166 bp in the 5′ region of a rearranged T‐cell receptor γ (TCRG) Vγ2 gene from the disease cells of a patient with T‐cell ALL. This abnormal rearrangement was not detected in a biopsy taken during clinical remission. Sequence analysis indicated that the deletion breakpoint occurred at a position immediately upstream of sequences found in the germline Vγ2 gene that are closely related to known heptamer and nonamer recombination signal sequences. Furthermore, the rearrangement was found to have non‐germline nucleotides (N‐region) in between otherwise intact V and J segments. These data indicate that this structure may be the result of an aberrant rearrangement event in common with the frequently occurring chromosomal abnormalities found in T‐cell ALL. This event could either be one directly associated with the leukaemic transformation or one occurring during normal lymphocyte development but which is coincidental with leukaemic transformation. This represents the first molecular genetic evidence for an abnormality specifically involving the TCRG locus in ALL.
Prognostic value of a novel circulating serum 90K antigen in HIV‐infected haemophilia patients
Authors: Longo G. , Natoli C. , Rafanelli D. , Tinari N. , Morfini M. , Rossi‐Ferrini P. , D'Ostilio N. , Iacobelli S.
Published: March 2008

Summary. We evaluated the prognostic role of a novel tumour‐associated antigen, termed 90K, in a cohort of HIV + asymptomatic haemophilia patients with known duration of seropositivity and median follow‐up of about 7 years. The circulating levels of 90K are higher in HIV + asymptomatic patients than HIV — controls. The antigen levels remain quite stable over time in non‐progressing patients, while they steadily rise in patients evolving to ARC/AIDS. Baseline high 90K levels are predictive of faster progression to ARC/AIDS and shorter survival. We conclude that an elevated 90K serum level is a predictor of poor prognosis in HIV+ asymptomatic haemophiliacs.
Life‐threatening thrombotic and haemorrhagic problems associated with silent myeloproliferative disorders
Authors: Vadher B. D. , Machin S. J. , Patterson K. G. , Sukhu C. , Walker H.
Published: March 2008

Summary. Myeloproliferative disorders are well recognized as being associated with haemorrhage and thrombosis. We describe two cases, one of life‐threatening haemorrhage and the other of thrombosis, in patients with normal peripheral blood counts and films, both of whom went on to develop overt manifestations of myeloproliferative disorders (CGL and essential thrombocythaemia) more than a year after their first presentation.
Respective effects of soluble interleukin‐1 receptor and tumour necrosis factor receptor on IL‐1 and TNF‐α‐induced DNA synthesis of common acute lymphoblastic leukaemia blasts in vitro
Authors: Carter Anna , Haddad Nuhad , Draxler Ilana , Israeli Ella , Raz Batya , Tatarsky Ilana
Published: March 2008

This study investigates the capacity of human recombinant interleukin‐1α (IL‐1α), IL‐1β and tumour necrosis factor‐α (TNF‐α) to induce DNA synthesis of highly purified blasts from nine adult common acute lymphoblastic leukaemias (cALL) in 7 d liquid culture.

IL‐1α, IL‐1β and TNF‐α stimulated 3H‐TdR uptake in leukaemic blasts in a dose‐dependent fashion. The IL‐1‐induced DNA synthesis of cALL cells could not be prevented by the addition of neutralizing antibodies against IL‐3, GM‐CSF, IL‐6 or TNF‐α. Similarly, the TNF‐α‐stimulated 3H‐TdR incorporation of leukaemic blasts was not affected by the addition of antibodies towards IL‐1α, IL‐1β, IL‐3, GM‐CSF or IL‐6. These observations suggest that IL‐1 as well as TNF‐α stimulated growth could not be attributed to the endogenous production of factors, corresponding to the antibodies used in these experiments. Both IL‐1 as well as TNF‐α mediate their action through interaction with specific cell surface receptors.

Recently two distinct types of IL‐1 receptors (IL‐1‐Rs), IL‐1R (p80) and IL‐1‐R (p65), as well as two distinct types of TNF‐receptors (TNF‐Rs), TNF‐R (p55) and TNF‐R (p75) have been identified. Both types of TNF‐Rs exist also in soluble forms (sTNF‐Rs), while soluble IL‐1‐Rs (sIL‐1‐Rs) have not yet been found naturally. In this study we show that sIL‐1‐R as well as sTNF‐R modulate the effects of their corresponding cytokine in a dose‐dependent bimodal fashion: at lower concentrations they augmented while at higher concentrations they inhibited the cytokine‐stimulated DNA synthesis of cALL blasts in vitro. It may therefore be concluded from this study that soluble receptors for both IL‐1 and TNF. at least in vitro, are functional and interfere with their corresponding cytokine bioactivity.
Immunotoxins containing saporin linked to different CD2 monoclonal antibodies: in vitro evaluation
Authors: Tazzari Pier Luigi , Bolognesi Andrea , de Totero Daniela , Lemoli Roberto M. , Fortuna Alessandra , Conte Roberto , Crumpton Michael J. , Stirpe Fiorenzo
Published: March 2008

In this study we describe immunotoxins prepared with different CD2 monoclonal antibodies (mAbs) and a ribosome‐inactivating protein, saporin. The CD2 immunotoxins were tested on different models. Anti‐CD2–saporin conjugates inhibited protein synthesis by a neoplastic CD2+ cell line (SKW‐3) and by an interleukin 2 dependent polyclonal CD2+ lymphoid cell culture (T lymphoblasts), with IC50s ranging from 10‐13m to 10‐11m (as saporin). Similar results were obtained with proliferation inhibition tests (3H‐thymidine incorporation) on phytohaemagglutinin (PHA) driven lymphoid cultures and on mixed lymphocyte culture activated lymphocytes. Moreover a CD2–ricin A chain conjugate was less effective than an analogous immunotoxin containing the same CD2 mAb and saporin in inhibiting lymphocyte proliferation induced by PHA (IC50 approximately 10‐9m as ricin A chain versus 10‐12m as saporin). The conjugates were not toxic on bone marrow stem cells. These results suggest that CD2–saporin immunotoxins could represent an effective tool for CD2+ lymphomas or leukaemias, and for T‐dependent immune disorders, such as transplanted organ rejection and graft‐versus‐host disease.
Influence of clinical status on the efficiency of stored platelet transfusion
Authors: Norol F. , Kuentz M. , Cordonnier C. , Beaujean F. , Haioun C. , Vernant J. P. , Duédari N.
Published: March 2008

The efficiency of stored platelet transfusion was evaluated in terms of clinical status in 141 thrombocytopenic patients. In a paired prospective study in which fresh platelets were used as controls, clinical efficiency was assessed on the basis of the ability to increase platelet count (recovery) and the time to the next transfusion (D). In 48 clinically stable patients, recovery of fresh and stored platelets was similar (47% and 41%, respectively) and the interval to the next transfusion was D4 and D3. In contrast, 27 patients who had bacterial infections showed significantly different recoveries (24%/5%) and the interval to the next transfusion was D3/D1 for fresh and stored platelets respectively. Similarly, in 16 patients who were treated concurrently with amphotericin B, 18 other patients with graft‐versus‐host disease, nine with splenomegaly and four with veno‐occlusive disease (VOD), fresh platelets performed better than stored platelets, showing recoveries of 27%/18%, 29%/15%, 16%/3% and 15%/2%. Furthermore, the need for retransfusion within 24 h was significantly increased with stored platelets. In 19 patients with anti‐HLA alloimmunization who were transfused with HLA‐matched fresh and stored apheresis platelet concentrate (APC), efficiency was similar (38%/36% and D4/D3). This study indicates that the storage induces an impressive decrease in the in‐vivo platelet recovery and survival in patients with certain clinical conditions.
Use of granulocyte colony‐stimulating factor (G‐CSF) for mobilizing peripheral blood stem cells: risk of mobilizing clonal myeloma cells in patients with bone marrow infiltration
Authors: Vora Ajay J. , Toh Cheng Hoc , Peel Janet , Greaves Michael
Published: March 2008

Peripheral blood stem cells have been used for autologous reconstitution of haemopoiesis after high dose cytotoxic therapy and produce similar disease response rates as autologous bone marrow transplants. Peripheral blood stem cell transplants are an especially attractive option for patients in whom marrow harvest is not feasible due to bone marrow damage or infiltration. Recombinant growth factors mobilize adequate numbers of stem cells from the marrow but their effect on tumour cell circulation kinetics is not known. We report a patient with multiple myeloma and bone marrow infiltration in whom the use of G‐CSF for stem cell mobilization led to release of plasma cells into the peripheral circulation and contamination of the stem cell harvest.
Expression of fos proto‐oncogene product by monoclonal antibody FO‐120 in smouldering adult T‐cell leukaemia (ATL)
Authors: Iwahashi Masahito , Otsuka Maki , Shimotakahara Shigemi , Uozumi Kimiharu , Hanada Shuichi , Arima Terukatsu , Matsumoto Makoto
Published: March 2008

We investigated the expression of c‐fos gene product in peripheral blood mononuclear cells of patients with smouldering adult T‐cell leukaemia (ATL) and healthy human T‐lymphotropic virus type‐I (HTLV‐I) carriers by an immunofluorescence assay, using a mouse monoclonal antibody (FO‐120) specific for fos gene product. Peripheral blood mononuclear cells derived from healthy HTLV‐I carriers were rarely positive for FO‐120, less than 2% of the cells weakly reacted with FO‐120, whereas positive cells were detected in more than about 10% of cells from patients with smouldering ATL.

FO‐120 appears to be a useful tool for detecting smouldering ATL in asymptomatic HTLV‐I infected people without using molecular techniques.