bluuebunny/crossref_metadata_2025 huggingface dataset:
Cultured marrow stromal cells express common acute lymphoblastic leukaemia antigen (CALLA): implications for marrow transplantation
Authors: Keating Armand , Whalen Connie K. , Singer Jack W.
Published: July 2010

Summary. This study demonstrates the presence of an antigenic determinant associated with the common acute lymphoblastic leukaemia antigen (CALLA), and presumably CALLA itself, on stromal cells in normal human long‐term marrow cultures by using two monoclonal anti‐CALLA antibodies, J‐5 and 24·1. Treatment of cultured stromal cells with antibody and complement resulted in the loss of most flat angulated cells and many of the fat‐containing cells. However, long‐term cultures were generated with cytotoxic antibody‐treated marrow buffy coat cells, and the stromal cells in these cultures were also CALLA‐positive. We conclude that CALLA‐bearing stromal cells arise from CALLA‐negative progenitors. CALLA therefore could be either a differentiation antigen acquired on mature marrow stromal cells or may arise as a proliferation‐dependent antigen. These studies suggest that the generation of long‐term cultures from cytotoxic antibody‐treated marrow may be an appropriate in vitro model for the functional assessment of such marrow prior to its use in autologous transplantation.
Molecular analysis of chromosome 22 breakpoints in adult Philadelphia‐positive acute lymphoblastic leukaemia
Authors: Kurzrock Razelle , Shtalrid Mordechai , Gutterman Jordan U. , Koller Charles A. , Walters Ronald , Trujillo Jose M. , Talpaz Moshe
Published: July 2010

Summary. The Philadelphia (Ph) translocation. t(9;22)(q34:q11). is found in the majority of patients with chronic myelogenous leukaemia (CML) as well as in approximately 20% of adult acute lymphoblastic leukaemia (ALL) patients. The chromosome 22 breakpoint in CML has been localized within a restricted 5.8 kb segment of DNA known as the breakpoint cluster region (bcr). To investigate the chromosome 22 breakpoint in ALL. we analysed five adult Ph‐positive ALL patients for bcr rearrangement. Rearrangement was detected within bcr in two patients. However, in one patient the break occurred 5’to the first exon of bcr and in two patients the bcr region was not involved. We conclude that the identical cytogenetic marker, t(9;22), may yield a different genomic configuration in ALL and CML.
Splenic blood flow and intrasplenic platelet kinetics in relation to spleen volume
Authors: Wadenvik Hans , Denfors Ingrid , Kutti Jack
Published: July 2010

The relationship between the splenic blood flow and the intrasplenic platelet kinetics on the one hand, i.e. the two factors which govern the size of the exchangeable splenic platelet pool, and the spleen size on the other were assessed in 21 patients afflicted with haematologic disorders and variable splenomegaly. The splenic blood flow and intrasplenic platelet kinetics were measured using111In‐labelled platelets and compartmental analysis of their equilibration between circulating blood and splenic pool; the spleen size was determined by scintigraphy using99mTc‐labelled stannous colloid. Significant correlations were recorded between the spleen size and the splenic platelet pool size (r= 0.76;P<0.001) and between the spleen size and the splenic blood flow (r= 0.56;P<0.01). Splenic perfusion decreased significantly with increasing spleen size, but there was no relationship between the spleen size and the intrasplenic platelet transit time. However, an association was present between splenic perfusion and intrasplenic platelet transit time (r=—0.44;P<0.05). It is concluded that the splenic blood flow is the major determinant of the size of the exchangeable splenic platelet pool in splenomegalic states, and that the determination of spleen size using99mTc‐scintigraphy gives a rough estimation of the pool size. Splenic perfusion appears to be one of the factors which determine the intrasplenic platelet transit time.
Assessment of bone marrow blood flow using positron emission tomography: no relationship with bone marrow cellularity
Authors: Martiat Ph. , Ferrant A. , Cogneau M. , Bol A. , Michel C. , Rodhain J. , Michaux J. L. , Sokal G.
Published: July 2010

Bone marrow blood flow has been assessed using positron emission tomography and the 15O‐labelled carbon dioxide steady‐state technique. The measurements were performed at the site of the posterior iliac crest. The bone marrow blood flow was 10.0 ml/min/100 cm3±3.0 (SD) in normal volunteers. It was markedly increased in patients with polycythaemia vera (26.9±4.6), chronic granulocytic leukaemia (25.2 ± 3.9) and myelofibrosis (35.1 ± 7.3). However, bone marrow blood flow did not differ from normal in patients with aplastic anaemia, chronic haemolysis or chronic lymphocytic leukaemia. There was no relationship between bone marrow cellularity and bone marrow blood flow. The data show that bone marrow blood flow is markedly elevated in polycythaemia vera, myelofibrosis and chronic granulocytic leukaemia and suggest that bone marrow cellularity is not a major factor in regulating bone marrow blood flow.
Marrow transplantation from HLA non‐identical family donors for the treatment of leukaemia: a pilot study of 15 patients using additional immunosuppression and T‐cell depletion
Authors: Cahn J. Y. , Herve P. , Flesch M. , Plouvier E. , Racadot E. , Vuillier J. , Montcuquet P. , Noir A. , Rozenbaum A. , Floris R. Leconte des
Published: July 2010

Fifteen patients with high‐risk leukaemia were given T‐cell depleted marrow transplants from HLA non‐identical related donors. They were treated with a combination of total body irradiation (TBI), high‐dose cytosine arabinoside (Ara‐C) and high‐dose melphalan in an attempt to prevent a host‐versus‐graft reaction. Antilymphocyte globulins were given prior to transplantation for additional immunosuppression to 13 patients and in‐vivo monoclonal antibody anti‐human LFA1 to two. Engraftment and chimaerism assessed by HLA typing were achieved in 14 patients. Seven developed acute graft‐versus‐host disease (two fatal), one failed to engraft. Six patients died in complete remission from cytomegalovirus (CMV) interstitial pneumonitis and three remain alive in complete remission 2, 3 and 13 months after transplant. We conclude that aggressive immunosuppression allows for sustained engraftment of T‐cell depleted HLA non‐identical marrow. The incidence and severity of GVHD are acceptable and CMV pneumonitis remains the major problem.
The interaction of alpha thalassaemia and sickle cell–beta° thalassaemia
Authors: Vyas P. , Higgs D. R. , Weatherall D. J. , Dunn D. , Serjeant B. E. , Serjeant G. R.
Published: July 2010

The effects of alpha thalassaemia on sickle cell–beta° thalassaemia have been studied by comparing haematological and clinical features in four subjects homozygous for alpha thalassaemia 2 (2‐gene group), 27 heterozygotes (3‐gene group), and 55 with a normal alpha globin gene complement (4‐gene group). Alpha thalassaemia was associated with significantly higher haemoglobin levels and lower reticulocyte counts independent of the presence of splenomegaly. Contrary to expectation, alpha thalassaemia was associated with small but significant increases in mean cell volume and mean corpuscular haemoglobin concentration. Splenomegaly at age 5 years and episodes of acute splenic sequestration were significantly more frequent in the 4‐gene group. There were no significant differences in painful crises, acute chest syndrome, or other clinical features.
Competition between recipient and donor cells after bone marrow transplantation for chronic myeloid leukaemia
Authors: Frassoni Francesco , Sessarego Mario , Bacigalupo Andrea , Strada Paolo , Repetto Mario , Miceli Salvatore , Occhini Domenico , Defferrari Raffaella , Marmont Alberto
Published: July 2010

The cytogenetic and clinical course of three patients allografted for Ph positive chronic myeloid leukaemia are reported. All patients had a peculiar pattern of relapse. Two out of three patients had donor marrow graft pretreated with monoclonal antibody for graft versus host prevention. The cytogenetic relapse was invariably associated with major morphological changes in the marrow indicating that these were also haematological relapses. However, no changes in the peripheral blood count were observed. When relapse occurred in these patients, Ph positive marrow metaphases and host red blood cells ranged from 75% to 100% of the total cell population: thereafter they spontaneously reverted to complete chimaerism. Therefore the presence of leukaemic cells even in considerable amount was not sufficient, per se, to prevail over normal marrow. In addition these observations indicate that relapse was not associated with elimination of the graft: while haemopoiesis was entirely of recipient origin the donor normal stem cells were present and vital although functionally silent. These data suggest that, although TBI remains the more effective tool for eradicating the majority of leukaemic cells, haemopoietic competition between host and donor marrow may have a major impact on leukaemic relapse.
No detectable malignant B cells in the peripheral blood of patients with multiple myeloma
Authors: Clofent G. , Klein B. , Commes T. , Ghanem N. , Lefranc M. P. , Bataille R.
Published: July 2010

Summary Previous studies have reported the presence of idiotypic B lymphocytes in the blood of patients with multiple myeloma (MM), suggesting that they may belong to the malignant clone. This led us to investigate by southern blot analyses the pesence of tumour‐specific immunoglobulin‐gene (Ig‐gene) rearrangements in the peripheral‐blood mononuclear cells of 21 MM patients. This method was shown to detect clonal cells when they represent as little as 2% of the cell population. B‐cell‐enriched fractions were also studied in nine cases. An occasional contamination by cruclatin malignant plasma cells was carefully evaluated using immunofluorescence. Clonal rearrangements were observed in only two cases, in which a contamination by myeloma cells was evident. In these cases the use of different endonucleases clearly demonstrated that these Igclonal rearrangements involved post‐switched cells. No clonal rearrangement was found when contamination by myeloma cells was absent. Our results demonstrate the absence of detectable B cells involved in the myeloma clone in the peripheral blood of patients with MM.
Human monoclonal anti‐D antibodies: II. THE RELATIONSHIP BETWEEN IgG SUBCLASS, Gm ALLOTYPE AND Fc MEDIATED FUNCTION
Authors: Kumpel B. M. , Wiener E. , Urlbaniak S. J. , Bradley B. A.
Published: July 2010

Summary. Eight monoclonal antibodies (mabs) to the Rh antigen D produced by Epstein‐Barr virus transformed Blymphoblastoid cell lines from two indivbduals have been compared for their behaviour in in vitro cell‐mediated assays. Three IgGl Glm(1.17) and two IgG3 G3m(21) mabs from one donor and three IgGl Glm(3) mabs from another were used. IgG3 anti‐D mabs induced greater adherence and phagocytosis of sensitized red cells by U937 monlocytes than IgGl anti‐D mabs or the polyclonal anti‐D. Minimum sensitization levels for rosetting and phagocytosis by 1393 7 monocytes were 2000 molecules IgG/cell for IgG3 and 5000 molecules/cell for IgGl mabs; maximum rosetting mediated by both IgGl and IgG3 mabs was obtained at 15000‐20000 molecules/cell. The IgG3 anti‐D mabs weire comparable to polyclonal anti‐D in mediating binding of sensitized red cells to γ‐interferon stimulated monocyte‐derived cultured macrophages and were markedly more effective than the IgGl anti‐D mabs. However, in lymphocyte ADCC assays, only anti‐D mabs which were IgGl Glm(3) were effective in mediating high levels of lysis of sensitized red cells, unlike the IgGl Glm (1.17) or IgG3 G3m(21) mabs. Minimum sensitization levels required for this lymphocyte‐mediated red cell lysis were found to be approximately 5000 molecules/cell with one IgGl Glm(3) mab; maximum lysis with this mab was obtained at 10000 molecules/cell. Polyclonal anti‐D containing both IgGl and IgG3 was effective in all three assays. These observations suggest that different isotypes and allotypes of anti‐D antibodies mediate red cell remo or destruction by monocyte or lymphocyte effector ce’through functionally dissimilar Fc receptor interactions.
Spontaneous NBT reduction by monocytes as a marker of disease activity in children with histiocytosis
Authors: Burgio G. Roberto , Aricó Maurizio , Marconi Massimo , Lanfranchi Arnalda , Caselli Désirée , Ugazio Alberto G.
Published: July 2010

In an attempt to define a biological marker of monocyte hyperactivation in the course of infantile histiocytosis, the spontaneous nitroblue tetrazolium (NBT) reduction assay was applied to monocytes from 13 children with Langerhans cell histiocytosis (LCH), familial haemophagocytic lymphohistiocytosis (FHL), juvenile xanthogranuloma or malignant histiocytosis. Significant increase in NBT reduction was observed in the patients with both active LCH and FHL in comparison with control subjects, who were either healthy or affected by different conditions. A close relationship between spontaneous reduction rate and clinical condition of the patients was evident in patients tested at diagnosis, during remission and during the course of disease reactivation. Interleukin‐1 (IL‐1) production by monocytes was also evaluated: the patients with LCH and FHL displayed a significant increase in in vitro IL‐1 production by lipopolysaccharide‐stimulated monocytes. In our experience the spontaneous NBT reduction assay was a sensitive, quite specific, low‐cost and reproducible test for the evaluation of children with histiocytosis. Useful information may be obtained at diagnosis but also during the clinical course of disease by using this marker of monocyte spontaneous activation.
A familial factor XIII subunit B deficiency
Authors: Saito Masanori , Asakura Hidesaku , Yoshida Tomotaka , Ito Keiko , Okafuji Kazuhiro , Yoshida Takashi , Matsuda Tamotsu
Published: July 2010

A 32‐year‐old woman with a bleeding tendency born of a consanguineous marriage, was found to have factor XIII subunit B deficiency. An abnormally low level of factor XIII activity was initially noticed and this finding led to further studies of the proband and her family. The notable features were: undetectable subunit B of factor XIII in the proband and her brother and reduced levels of subunit B, 34–52%, in her parents and children. The proband's brother had a markedly decreased level of subunit A protein. The level of factor XIII subunit A in platelets of the proband was normal. The half‐life of subunit A determined from the disappearance curve of infused factor XIII subunit A concentrate was approximately 3 d and this is the shortest estimate of the half‐life of factor XIII to date. From these results, it is suggested that subunit A is unstable in plasma deficient in subunit B and subunit B stabilizes the A protein.

This is the first report of congenital deficiency of factor XIII subunit B and this disorder is thought to be inherited as an autosomal recessive trait.
Lack of IL‐1 secretion from human myeloma cells highly purified by immunomagnetic separation
Authors: Borset Magne , Helseth Eirik , Naume Bjørn , Waage Anders
Published: July 2010

Summary. This paper describes a method for purification of human myeloma cells. Mononuclear cells from six bone marrow samples and one pleural fluid sample from multiple myeloma patients were incubated with B‐B4. a monoclonal antibody that is specific for plasma cells, and the B‐B4+ cells were isolated using monosized magnetic beads coated with sheep anti‐mouse Ig. With this positive selection method it was possible to achieve primary cultures with more than 99% myeloma cells. The average viability of these cultures was 81%. The B‐B4 antibody did not alter proliferation or cytokine production of the myeloma cell lines U‐266 and JJN‐3. The B‐B4+ myeloma cultures did not produce IL‐1 and made only small amounts of IL‐6 (<93 pg/ml), whereas the cells remaining after extraction of the B‐B4+ cells produced IL‐1 (89–350 pg/ml) and large amounts of IL‐6 (520–17000 pg/ml). This indicates that the B‐B4+ myeloma cells are not directly responsible for the overproduction of these cytokines in multiple myeloma. This separation technique gave higher purity of myeloma cells than has been previously reported for any negative selection method and is recommended when high culture purity is of critical interest.
Fibrin glue in surgery: frequent development of inhibitors of bovine thrombin and human factor V
Authors: Bänninger Hans , Hardegger Theres , Tobler Andreas , Barth Alain , Schupbach Peter , Reinhart Walter , Lammle Bernhard , Furlan Miha
Published: July 2010

Summary. We report on a 34‐year‐old woman whose plasma showed a marked prolongation of thrombin time (TT) (< 200 s) using bovine thrombin. The patient had previously been exposed twice to topical bovine thrombin contained in fibrin glue during cardiac surgery. TT was normal when human thrombin was used as reagent. The patient's purified IgG reacted with bovine prothrombin and bovine thrombin in immunoblotting studies but showed virtually no cross‐reaction with human thrombin. In addition, following surgery, factor V clotting activity (FV:C) was reduced to 9% of normal. The inhibitor of bovine thrombin persisted over a period of more than a year, while the level of FV:C progressively returned to normal within this time period.

Development of thrombin and FV:C inhibitors was also investigated in plasma of 34 consecutive patients who had undergone either cardiac surgery or neurosurgery with use of fibrin glue containing bovine thrombin. Eleven of 24 patients after cardiac surgery and two of 10 patients after neurosurgery presented with TT ≥25 s (normal plasma 15s). Two patients had been re‐exposed to fibrin glue during cardiac re‐operation and showed markedly prolonged TT (> 60 s). All 13 patients who had acquired a thrombin inhibitor also had low FV:C activity (10–60% of normal plasma), whereas FV:C activity remained in the normal range in the 21 patients with normal TT. Our findings indicate that development of inhibitors of bovine thrombin as well as co‐immunization to factor V occurs frequently and is associated with the amount of applied fibrin glue and with the type of operation. Re‐exposure to fibrin glue seems to enhance formation of inhibitors of bovine thrombin and human factor V.
Sustained trilineage response in a patient with ALG‐resistant severe aplastic anaemia after treatment with G‐CSF, erythropoietin and cyclosporin A: association of recovery with marked elevation of serum alkaline phosphatase
Authors: Weide R. , Lyttelton M. , Samson D. , Görg C. , Köppler H. , Pflüger K. H. , Havemann K.
Published: July 2010

Summary. Aplastic anaemia is characterized by multilineage bone marrow failure resulting in pancytopenia. We have successfully treated a young woman with severe aplastic anaemia (SAA) who was resistant to antilymphocyte globulin (ALG) and corticosteroids, with a combination therapy consisting of erythropoietin. cyclosporin A and granulocyte‐colony stimulating factor (G‐CSF). The patient received erythropoietin and CSA for a period of 10 months without success before G‐CSF treatment was started. After 6 weeks of G‐CSF therapy she responded with a sustained trilineage recovery. This suggests that immunosuppression together with haemopoietic growth factors may be an effective treatment in patients with SAA who are ALG resistant and cannot be treated by BMT.
Remobilization of patients who fail to achieve minimal progenitor thresholds at the first attempt is clinically worthwhile
Authors: Watts M. J. , Ings S. J. , Flynn M. , Dodds D. , Goldstone A. H. , Linch D. C.
Published: July 2010

A significant proportion of previously treated patients fail to mobilize sufficient stem/progenitor cells to enable high‐dose therapy and peripheral blood stem cell transplantation to be performed. In this study, the value of remobilizing such patients has been evaluated in 20 patients who all failed to achieve progenitor yields of 1 × 106/kg CD34+ cells and 1 × 105/kg granulocyte–monocyte colony‐forming units (GM‐CFCs) at the first attempt. Most patients remained relatively poor mobilizers at the second mobilization, but the yield of CD34+ cells and GM‐CFCs on the first apheresis was significantly greater with the second mobilization than the first. A total yield (all aphereses from both mobilizations) of > 1 × 106/kg CD34+ cells and > 1 × 105/kg GM‐CFCs was achieved in 14 out of 20 patients. Seven patients have received high‐dose therapy with stem cell infusion; neutrophil recovery was rapid in all patients and platelet independence occurred in < 21 d in five out of seven patients. We conclude that remobilization is worth considering in those patients in whom a chemotherapy‐free interval of several months is possible.
Expression of interleukin 3 and granulocyte–macrophage colony‐stimulating factor receptor common chain βc, βIT in normal haematopoiesis: lineage specificity and proliferation‐independent induction
Authors: Militi Stefania , Riccioni Roberta , Parolini Isabella , Sposi Nadia Maria , Samoggia Paola , Pelosi Elvira , Testa Ugo , Peschle Cesare
Published: July 2010

Interleukin 3 (IL‐3), granulocyte–macrophage colony‐stimulating factor (GM‐CSF) and interleukin 5 (IL‐5) exert their biological activities through interaction with cell‐surface receptors that consist of two subunits, a specific α subunit and a common β transducing subunit (βc). We have evaluated the expression of βc on purified haematopoietic progenitor cells (HPCs) induced to unilineage differentiation/maturation through the erythroid (E), granulocytic (G), megakaryocytic (Mk) or monocytic (Mo) lineage. HPCs displayed low βc expression, which increased during the initial stages of HPC differentiation along the E, G, Mo or Mk lineages. At later stages of differentiation, βc chain expression increased in both G and Mo lineages, was expressed at low levels in the Mk lineage and declined to undetectable levels in the E lineage. Analysis of the full‐length βc and intracytoplasmically truncated βc (βIT) mRNAs showed that the former was predominant in the G and Mo lineages, whereas the latter was prevalent in the E and Mk lineages. The βc induction takes place even in the absence of cell cycling. Thus, incubation of HPCs with graded amounts of IL‐3 showed that the initial induction of βc expression is unrelated to cell proliferation. Furthermore, circulating monocytes and granulocytes exhibit a low level of βc expression that is greatly stimulated following incubation with either IL‐3 or GM‐CSF.
A new polymorphism in the human factor VIII gene: implications for linkage analysis in haemophilia A and for the evolution of int22h sequences
Authors: Bowen Derrick J. , De Brasi Carlos D. , Larripa Irene B. , Collins Peter W.
Published: July 2010

A new polymorphism in the human factor VIII gene has been localized and characterized. It is a biallelic, single nucleotide polymorphism located in intron 22 of the gene, within the 9·5 kb int22h‐1 segment. The allelic forms are G (frequency 0·65) and A (frequency 0·35), giving a predicted rate of heterozygosity of 0·46. The polymorphism occurs within a CG dinucleotide and affects an MspI site (CCGG). Int22h‐1 is duplicated twice extragenically at Xq28; both extragenic copies (int22h‐2 and ‐3) are also polymorphic with respect to MspI. Investigation of 156 MspI [–] alleles, comprising 30 intragenic and 126 extragenic sites, indicated that all were due to A alleles and none had arisen by C to T transition within the CG dinucleotide. The intragenic MspI site (designated MspI A) is located 737 bases downstream of a previously described XbaI restriction fragment length polymorphism. Despite their close proximity, the polymorphisms are not in complete linkage disequilibrium; haplotype analysis in 85 factor VIII genes from a Caucasian population predicts an informativity of approximately 60% in linkage studies using both, compared with an informativity of approximately 47% in studies using either on its own.
Intercellular communication between bone marrow stromal cells and CD34+ haematopoietic progenitor cells is mediated by connexin 43‐type gap junctions
Authors: Dürig Jan , Rosenthal Christoph , Halfmeyer Katja , Wiemann Martin , Novotny Jürgen , Bingmann Dieter , Dührsen Ulrich , Schirrmacher Karin
Published: July 2010

The existence of functional gap junctions between haematopoietic progenitor cells (HPCs) and stromal cells of the haematopoietic microenvironment in the human system is a controversial issue. Primary CD34+ HPCs isolated from leukapheresis products were co‐incubated with the human fibroblastoid bone marrow stromal cell line L87/4 in short‐term liquid culture. Using the highly sensitive double whole‐cell patch‐clamp technique, we found that the majority (91%) of CD34+ HPCs are electrically coupled to L87/4 cells. Importantly, efficient coupling was observed within 1 h of the attachment of CD34+ HPCs to plastic adherent L87/4 cells. By comparison, homologous cell pairs formed by L87/4 cells exhibited a significantly higher electric coupling. Analysis of single‐channel conductances revealed an electric profile characteristic of connexin 43 (Cx43)‐type gap junctions for both homologous and heterologous cell pairs. The Cx phenotype was confirmed using Cx43‐specific monoclonal antibodies in a flow cytometric assay and reverse transcription polymerase chain reaction (RT‐PCR) for the detection of Cx43 mRNA. Finally, the electrophysiological studies were complemented by dye‐transfer experiments using the recently described ‘parachute’ technique that allows the monitoring of dye diffusion without disruption of the plasma membrane. Taken together, our data indicate that functional Cx43‐type gap junctions exist between stromal cells and immature HPCs and, thus, may provide an important regulatory pathway in haematopoiesis.
Parvovirus B19 transmitted by bone marrow
Authors: Heegaard Erik D. , Laub Petersen Bodil
Published: July 2010

We describe a case of symptomatic parvovirus B19 infection transmitted by bone marrow (BM). The infection caused prolonged anaemia, thrombocytopenia, arthralgia and erythema infectiosum in a 16‐year‐old girl with acute myeloid leukaemia receiving a BM transplant (BMT). The BM donor was a 19‐year‐old asymptomatic brother who had parvovirus B19 viraemia at the time of BM harvest. Sequencing of the VP2 gene from the patient and the donor showed a perfect match of DNA sequences, confirming the mode of transmission. Parvovirus B19 represents a potential complicating factor in patients undergoing BMT, but screening by polymerase chain reaction (PCR) of donor BM may reduce the risk of infection.
An Asian variant of intravascular large B‐cell lymphoma: clinical, pathological and cytogenetic approaches to diffuse large B‐cell lymphoma associated with haemophagocytic syndrome
Authors: Murase Takuhei , Nakamura Shigeo , Kawauchi Kiyotaka , Matsuzaki Hiromitsu , Sakai Chikara , Inaba Tohru , Nasu Kaori , Tashiro Kazuhiro , Suchi Taizan , Saito Hidehiko
Published: July 2010

Diffuse large B‐cell lymphoma with haemophagocytic syndrome (BCL‐HS) has been reported mainly in Asia and is regarded as a distinct variant of intravascular lymphoma (IVL). However, it is unclear whether all cases of BCL‐HS fall within the framework of IVL and available clinical information is limited. We analysed 25 cases with BCL‐HS, including 11 autopsied cases (median, 66 years; male–female ratio, 1·1:1). The patients presented with fever, anaemia, thrombocytopenia, hepatosplenomegaly, haemophagocytosis, bone marrow invasion, respiratory disturbance and disseminated intravascular coagulopathy, but usually lacked lymphadenopathy, mass formation, neurological abnormalities and skin lesions. The clinical course was aggressive with a median survival of 7 months. The morphological findings were uniform: large lymphoid cells infiltrated vessels and/or sinusoids of the liver, marrow, lung, kidney and other organs. They were positive for CD19, CD20, CD79a and HLA‐DR, but negative for CD10, CD23 and CD30. CD5 was positive in five out of 17 cases. Our critical review indicates that BCL‐HS is the equivalent of the Asian variant of IVL.
Affordable CD4+ T‐cell counts on ‘single‐platform’ flow cytometers I. Primary CD4 gating
Authors: Janossy G. , Jani I. , Göhde W.
Published: July 2010

Here, we demonstrate the flow cytometric concept of ‘primary CD4 gating’ utilizing three different CD4 monoclonal antibodies (mAbs) conjugated with five different fluorochromes. CD4+ lymphocytes were defined by an autogate in a single histogram of CD4 fluorescence intensity (FI) (y‐axis) vs. side light scatter (x‐axis). A wide range of absolute counts for > 600 individuals, including HIV+ patients, were compared with those obtained by ‘state‐of‐the‐art’ single‐platform flow cytometers such as the volumetric Ortho CytoronAbsolute and the Becton Dickinson FACSCalibur using TruCount beads. The correlation between CD4 counts obtained with primary CD4 gating and the full test panel on the Ortho Cytoron was excellent (R2 = 0·999). Bland–Altman statistics showed a mean difference of −2 cells/mm3[confidence interval (CI) 95% = −3 to −1; limits of agreement −27 to +23]. In addition to absolute CD4 counts, CD4% values and CD4/CD8 ratios are also frequently requested. To obtain these, lymphocytes need to be counted using scatter gates, and a second tube stained with a CD8 mAb to count CD8++ lymphocytes can be incorporated. We conclude that primary CD4 gating on single‐platform volumetric flow cytometers is one of the most economical and flexible technologies for routine cost‐conscious service work, particularly during the follow‐up of patients undergoing anti‐HIV therapy and/or vaccination in the developing world.
Additional chromosomal abnormalities in patients with acute promyelocytic leukaemia (APL) do not confer poor prognosis: results of APL 93 trial
Authors: de Botton Stéphane , Chevret Sylvie , Sanz Miguel , Dombret Hervé , Thomas Xavier , Guerci Agnès , Fey Martin , Rayon Consuelo , Huguet Françoise , Sotto Jean‐Jacques , Gardin Claude , Cony Makhoul Pascale , Travade Philippe , Solary Eric , Fegueux Nathalie , Bordessoule Dominique , San Miguel Jesus , Link Harmut , Desablens Bernard , Stamatoullas Aspasia , Deconinck Eric , Geiser K , Hess U , Maloisel Fréderic , Castaigne Sylvie , Preudhomme Claude , Chomienne Christine , Degos Laurent , Fenaux Pierre , the European Apl Group for
Published: July 2010

In spite of the recent improvement in the outcome of acute promyelocytic leukaemia (APL) with treatment combining all trans retinoic acid (ATRA) and chemotherapy (CT), some patients with this disease still have a poor outcome. The prognostic significance of chromosomal abnormalities in addition to t(15;17) in APL is uncertain. We examined the prognostic significance of secondary chromosomal changes in 292 patients included in a European trial who were treated with ATRA and CT. The incidence of chromosomal abnormalities in addition to t(15;17) was 26% and trisomy 8 was the most frequent secondary change (46% of the cases with secondary changes). No significant differences were seen with regard to age, sex, initial white blood cell count, % of circulating blasts, platelet count, fibrinogen level and incidence of microgranular variants between patients with or without additional rearrangements. Outcome was also similar between patients with t(15;17) alone and patients with t(15;17) and other clonal abnormalities for complete remission (92% vs. 93% respectively), event‐free survival at 2 years (76·1% vs. 78·1% respectively), relapse at 2 years (16·7% vs. 11·6% respectively) and overall survival at 2 years (79·9% vs. 79·5% respectively). Analysis according to the type of induction treatment (ATRA followed by CT or ATRA plus CT) or the type of maintenance treatment (with ATRA, low‐dose CT or both) also failed to show any difference between the two groups. Thus, in a large cohort of APL patients treated with ATRA and CT, additional chromosomal abnormalities had no impact on prognosis.
Recombinant VIIa concentrate in the management of bleeding following prothrombin complex concentrate‐related myocardial infarction in patients with haemophilia and inhibitors
Authors: Hough R. E. , Hampton K. K. , Preston F. E. , Channer K. S. , West J. , Makris M.
Published: July 2010

Prothrombin complex concentrates (PCCs) and, more recently, activated prothrombin complex concentrates (APCCs), are widely used for the treatment of active bleeding in haemophiliacs with inhibitors. Myocardial infarction (MI), associated with the use of these concentrates, is a well‐recognized, but uncommon, complication. We review the 14 previous cases published in the literature and describe two additional patients. MI related to the use of activated and non‐activated PCCs predominantly affects young patients who often have no preceding history of, or risk factors for, MI and tends to be associated with large cumulative doses of concentrate. The most frequent pathological finding is myocardial haemorrhage, with no evidence of coronary artery atheroma or thrombosis. The management of further bleeding in these patients is difficult. We have safely used recombinant factor VIIa to treat bleeding in the immediate and long‐term period following PCC‐related MI.
Haematologists' approaches to the management of adolescents and young adults with acute leukaemia
Authors: Benjamin S. , Kroll M. E. , Cartwright R. A. , Clough J. V. , Gorst D. W. , Proctor S. J. , Ross J. R. Y. , Taylor P. R. A. , Wheatley K. , Whittaker J. A. , Stiller C. A.
Published: July 2010

Approaches to the management of adolescents and young adults with acute leukaemia were investigated by sending a questionnaire to hospitals identified as having diagnosed or treated patients aged 15–29 years. The responses demonstrated the types of hospital treating these patients, the haematologists' perceived practice for entry of patients to Medical Research Council (MRC) leukaemia trials and reasons for non‐entry. Data were linked to MRC trials data to determine the proportion of patients aged 15–29 years at diagnosis in responding hospitals actually treated in MRC leukaemia trials in the 5 years preceding the questionnaire. Eighty‐two per cent of haematologists stated that they entered patients ‘always’ or ‘whenever possible’ for acute myeloid leukaemia (AML) and 76% for acute lymphoblastic leukaemia (ALL), but actual entry rates from the study hospitals were 46% of 239 AML patients and 36% of 182 ALL patients. The reasons most commonly reported for not entering eligible patients to national leukaemia trials were clinician preference for one arm of an MRC trial, a regional study or non‐trial protocol, and concern about workload and ethical approval.
Prevalence of erythrocyte haemoglobin H inclusions in unselected patients with clonal myeloid disorders
Authors: Steensma David P. , Porcher Julie C. , Hanson Curtis A. , Lathrop Cynthia L. , Hoyer James D. , Lasho Terra A. , Tefferi Ayalew , Higgs Douglas R.
Published: July 2010

Patients with clonal myeloid disorders, especially myelodysplastic syndromes (MDS), may acquire α‐thalassaemia. To estimate the prevalence of this erythrocyte phenotype, we examined brilliant cresyl blue‐stained blood smears from 201 patients with neoplastic myeloid disorders and 282 controls (195 non‐clonal anaemia, 62 with medical illnesses without anaemia and 25 healthy persons). Haemoglobin H inclusions were detected in 8/100 patients with MDS (8%) and 2/81 (2.5%) patients with myeloproliferative disorders, but in none of the acute leukaemia patients or controls. We conclude that the emergence of thalassaemic clones may be relatively common in the disordered marrow milieu of MDS.
Beneficial effects of the thymic hormone preparation thymoxtimulin in patients with defects in cell-mediated immunity and chronic purulent rhinosinusitis. A double-blind cross-over trial on improvements in monocyte polarization and clinical effects
Authors: TAS M , LEEZENBERG J A , DREXHAGE H A
Published: July 2010

Twenty patients with chronic purulent rhinosinusitis were treated with TP-1 (Serono; 1 mg/kg body weight), in a double-blind cross-over trial. TP-1 was administered by daily i.m. injections for the first 14 days followed by two injections/week for 6 further weeks. The patients were immunologically special in that they had defects in their cell-mediated immune system. Fourteen showed a decreased chemotactic responsiveness of their peripheral blood monocytes as measured in the polarization assay. This defective function can probably be ascribed to the presence in serum of low molecular weight factors (LMWFs; < 25 kD). As reported earlier, this factor shows a structural homology to the envelope protein of murine and feline leukaemia virus (P15E). Thirteen patients showed a defective delayed-type hypersensitivity (DTH) skin test reactivity towards candidin and/or streptokinase-streptodornase (Sk/Sd) antigen. 14 had a defective MIF production from their peripheral blood lymphocytes towards candidin, Sk/Sd and/or Haemophilus influenzae antigen. Eighteen patients completed the TP-1 trial and showed clinical improvements: 12 out of 15 were feeling better during TP-1 therapy and the nasal mucosa showed on inspection absent mucopurulent secretion in 13 patients. Positive bacterial culture rates for the nose decreased from 14 out of 16 to five out of 15. Placebo treatment had no significant effects. The clinical improvements were accompanied by a better performance of the cell-mediated immune system: the most significant effects were recorded in the monocyte polarization assay. The suppressive P15E-like LMWFs in serum clearly decreased during TP-1 treatment. In vitro TP-1 neutralized the immunosuppressivc effect of the LMWFs. The restoring effects of TP-1 on monocyte polarization and its neutralizing activity of P15E-like LMWFs could explain the beneficial effects of thymic hormone treatment reported in adults with clinical signs of immunodeficiency in the presence of a full T cell repertoire.
Specific lysis of mycobacterial antigen-bearing macrophages by class II MHC-restricted polyclonal T cell lines in healthy donors or patients with tuberculosis
Authors: KUMARARATNE D S , PITHIE A S , DRYSDALE P , GASTON J S H , KIESSLING R , ILES P B , ELLIS C J , INNES J , WISE R
Published: July 2010

The cytolytic capacity of mycobacterial antigen-stimulated peripheral blood mononuclear cells, from healthy Mantoux-positive volunteers and from patients with tuberculosis was investigated. Polyclonal T cell lines induced by 7 days of stimulation in vitro with PPD or a sonicate of Mycobacterium tuberculosis lysed both autologous macrophages and Epstein Barr virus (EBV) transformed B cell lines which had been pulsed with mycobacterial antigens, to a greater extent than unpulsed target cells or target cells pulsed with an irrelevant antigen (streptokinase/streptodornase). The killing of mycobacterial antigen-pulsed macrophages and EBV-transformed B cell line targets was inhibited by monoclonal antibodies to MHC class II antigens but not by antibodies directed against MHC class I antigens. PPD-pulsed EBV-transformed lymphoblastoid cell lines (LCL) competitively inhibited the killing of mycobacterial antigen-pulsed macrophages. whereas natural killer (NK) sensitive K562 cells (with or without antigen pulsing) did not inhibit mycobacterial antigen-dependent cytolysis of macrophages. Patients with tuberculosis showed a spectrum of mycobacterial antigen-induced cytolytic capacity. Those with extensive tissue necrosis (e.g. cavitatory pulmonary tuberculosis or caseous, extrathoracic tuberculosis) had high levels while patients with disseminated (miliary) tuberculosis or disease refractory to treatment showed little evidence of mycobacterial antigen induced cytotoxicity. The ability of mycobacterial antigenstimulated lymphoblasts to kill specific antigen-pulsed autologous macrophages was not significantly different between healthy donors and patients with tuberculosis. However, the‘mycobacterial antigen-specific’ component of this cytolysis was significantly deficient (P < 0·01) in patients. We conclude that mycobacterial antigen-specific cytotoxic T cell responses may play a significant part in the immune response to mycobacterial infection.
Soluble CD8 in patients with rheumatic diseases
Authors: SYMONS J A , WOOD N C , GIOVINE F S DI , DUFF G W
Published: July 2010

An ELISA was used to measure soluble CD8 (sCD8) in the sera and synovial fluids (SF) of patients with rheumatic diseases. Patients with rheumatoid arthritis (RA) had raised levels of sCD8 both in their sera and in their SF compared with patients with osteoarthritis and age-matched healthy controls. In individual RA patients, serial serum sCD8 levels initially fell and then rose preceding clinical improvement. In four patients where serum sCD8 levels rose and clinical improvement occurred, subsequent spontaneous decreases of serum sCD8 level preceded increased clinical disease activity by up to 2 weeks. In general, RA SF mononuclear cells (SFMNC) spontaneously produced high levels of sCD8. In contrast, autologous peripheral blood MNC only produced comparable levels after mitogenic stimulation. Incubation of SFMNC with increasing concentrations of human recombinant tumour necrosis factor alpha resulted in a dose-dependent potentiation of sCD8 release into the supernatant. There was an inverse relationship between the ability of SFMNC to release sCD8 and soluble interleukin-2 receptor, indicating that the CD8+ T cell population may play an important immunoregulatory role in RA.
Immunoglobulin class and subclass restriction of autoimmune responses in secondary syphilis
Authors: VERSALOVIC J , NASH Z-D , CARINHAS R , MUSHER D M , BAUGHN R E
Published: July 2010

The immunoglobulin (Ig) class and IgG subclasses of autoantibodies to commercial VDRL antigen, creatine kinase (CK), and fibronectin (Fn) in the sera of patients with various stages of syphilis were quantified using solid-phase radioimmunoassays (RIA) and ELISA. Sera from patients with active secondary syphilis, initially positive for anti-Fn and anti-CK autoantibodies by RIA, were reevaluated by ELISA using monoclonal antibodies (MoAb) for detection of human Ig class and subclass responses. Results of these assays revealed that anti-Fn and anti-CK responses were not only IgG in nature, but dramatically skewed to IgGl and IgG3 subclasses. While the restricted, co-expression of these isotypes seemingly paralleled anti-treponemal activity, inverse relationships actually existed between the subclass responses to Fn and those to Treponema pallidum. In contrast, anti-VDRL were predominantly IgM in 17 of 22 patients. Of those sera exhibiting detectable anti-VDRL IgG activity, responses appeared to be restricted to IgG1. These results suggest that different control mechanisms may be responsible for regulation of the various autoantibody responses expressed during syphilitic infection.
The CD8+ Leu-7+ subset of T cells in Crohn's disease: distinction between cytotoxic and covert suppressor functions
Authors: DEEM R L , TARGAN S R , NIEDERLEHNER A , SHANAHAN F
Published: July 2010

An expanded T cell subpopulation (CD8 + Leu-7+) has previously been reported in the peripheral blood of patients with Crohn's disease. This subpopulation of T cells was associated with a‘covert suppressor’ function, particularly in patients with mild/early Crohn's disease, suppressing immunoglobulin production in vitro when cultured in the presence of pokeweed mitogen. T cells with the same CD8 + Leu-7+ phenotype have also been shown to exhibit non-major histocompatability complex-restricted cytotoxicity when triggered by anti-CD3 antibodies, and this cytotoxic activity has also been shown to be elevated in patients with Crohn's disease. Because cytotoxic cells can have immunoregulatory properties, we investigated the possible relationship between the cytotoxic and‘covert suppressor’ functions of the CD8+ Leu-7+ subset of T lymphocytes in patients with mildly active Crohn's disease. Although the correlation between T cell cytoloxic activity and the CD8+ Leu-7+ cells was confirmed, no evidence for covert suppressor activity was found; there were no significant differences between the amount of IgM secreted by B cells from normal subjects and patients with Crohn's disease when cultured with T cells at increasing T: B ratios. In addition, IgM production by peripheral blood B cells did not correlate with either the number of CD8+ Leu-7+ cells or with the level of cytotoxic T cell activity. Furthermore, when B cells and CD4+ T cells were co-cultured with increasing numbers of CD8+ T cells, there was no evidence for excessive suppressor T cell activity in Crohn's disease. Although some patients exhibited low levels of IgM production, this was due to diminished B cell function, rather than excessive T suppressor activity or defective T helper activity. We conclude that the CD8+ Leu-7+ T cell subset is associated with cytotoxic but not with enhanced or covert suppressor activity in Crohn's disease. The previously described covert suppressor function attributed to cells with this phenotype in Crohn's disease was not found to account for diminished B cell responsiveness in vitro and is unlikely to be of major pathophysiologic significance in the majority of patients.
Detection of auto-anti-idiotypic antibodies toLol p I(rye I) IgE antibodies in human sera by the use of murine idiotypes: levels in atopic and non-atopic subjects and effects of immunotherapy
Authors: HÉBERT J , BERNIER D , MOURAD W
Published: July 2010

Anti-idiotypic antibodies (anti-Id Abs) are involved in the regulation of a number of immune responses including the IgE antibody production. In atopic patients, the increased synthesis of IgE antibodies could be related to a defective production of regulatory anti-Id Abs. In the present study, we first developed a sensitive assay for measuring the levels of anti-Id Abs directed against antibodies specific for Lol p I, the major allergenic determinant of Lolium perenne (rye grass). In this assay, we used previously described murine monoclonal anti-Lol p I antibodies that were shown to share epitopic specificities with human anti-Lol p I IgE and IgG antibodies, thus short-cutting the need for purification of F(ab′)2 fragments of human IgG Abs and insuring optimal specificity and sensitivity. Levels of anti-Id Abs against two anti- Lol p I monoclonal antibodies (290A-167. 348A-6) were higher in normal volunteers than in untreated atopic patients. Specific immunotherapy increased the levels of anti-Id Abs to those of normal volunteers. These observations suggest a role for the Id-anti-Id network in the regulation of IgE antibody production.