bluuebunny/crossref_metadata_2025 huggingface dataset:
Two human myeloma cell lines, amylase‐producing KMS‐12‐PE and amylase‐non‐producing KMS‐12‐BM, were established from a patient, having the same chromosome marker, t(11;14)(q13;q32)
Authors: Ohtsuki Takemi , Yawata Yoshihito , Wada Hideho , Sugihara Takashi , Mori Masaharu , Namba Masayoshi
Published: March 2008

Two human myeloma cell lines, KMS‐12‐PE and KMS‐12‐BM, were established from a 64‐year‐old woman with a non‐producing type of multiple myeloma. The KMS‐12‐PE line originated from the pleural effusion and the KMS‐12‐BM from the bone marrow. These two lines showed the same chromosome marker, t(11;14)(q13;q32). However, their phenotypes of surface markers differed from each other. KMS‐12‐BM cells were positive to CD20, CD38 and PCA‐1, showing the plasmacytoid (immature plasma cell) stage of B‐cell differentiation, while KMS‐12‐PE cells were positive to CD38 and PCA‐1, but not to CD20, indicating the terminal differentiated stage of B‐cells. As seen in the pleural effusion of the patient, KMS‐12‐PE cells ectopically produced a salivary type of amylase, but KMS‐12‐BM cells did not. Interestingly, the chromosome abnormality of del(1)(p22→pter) near the region of Ip21, where the amylase gene was assigned, was noticed in as many as 76% of KMS‐12‐PE cells.
Identification of H‐ and L‐ferritin subunit binding sites on human T and B lymphoid cells
Authors: Anderson G. J. , Faulk W. Page , Arosio P. , Moss D. , Powell L. W. , Halliday J. W.
Published: March 2008

Monoclonal antibodies to the H and L subunits of ferritin were used in indirect immunofluorescent studies to investigate the subunit specificity of ferritin binding to transformed T and B lymphoid cell lines. Pre‐incubation of cells with recombinant H ferritin followed by antibodies to the H subunit produced a labelling pattern in which all cells showed strong fluorescent stippling. Corresponding experiments with liver ferritin and an antibody to the L subunit revealed that most cells were lightly stippled, but 3–5% of the cells were intensely labelled. These two labelling patterns were distinct as assessed by cross‐blocking experiments with recombinant H and liver ferritins. These results imply that different binding sites exist for each subunit.
An immunological cross‐reactant of erythropoietin in serum which may invalidate EPO radioimmunoassay
Authors: Cotes P. Mary , Tam R. C. , Reed P. , Hellebostad M.
Published: March 2008

Atypically high radioimmunoassay estimates of erythropoietin in serum samples from two out of a series of 130 normal Norwegian children have been found to be invalid. Invalidity was due to non‐parallelism of radioimmunoassay (RIA) response dilution lines given by these two sera when compared with the second International Reference Preparation (IRP). In contrast, immunoreactive erythropoietin in other sera usually shows parallelism with the IRP. Fractionation by gel filtration of the sera showing non‐parallelism in the RIA showed them to contain an erythro‐poietin cross‐reactant which was eluted before both an 125I recombinant human erythropoietin marker and normal erythropoietin.

In family studies, non‐parallelism of RIA response dilution lines and the large molecular size erythropoietin cross‐reactant was found in serum from one sibling but not in five other family members of one of the two index cases.

Radioimmunoassays based on two different antisera differed in recognition of the abnormal cross‐reactant of erythropoietin.
Enhancement of monoclonal antibody dependent cell mediated cytotoxicity by IL2 and GM‐CSF
Authors: Bianchi Alessandra C. , Heslop Helen E. , Veys Paul , Macey Marion , Holland Mark , Prentice H. Grant , Brenner Malcolm K.
Published: March 2008

Rodent monoclonal antibodies (MAb) directed against cells of the immune system may be used in vivo for applications including conditioning prior to marrow transplantation and treatment of lymphoid malignancies. Although some MAb may lyse targets by complement fixation, MAb dependent cell mediated cytotoxicity (MAb‐DCC) appears to be an important additional effector mechanism. We have investigated the cellular basis of the phenomenon and the response of the effector cells to recombinant cytokines in an attempt to maximize the efficacy of MAb‐DCC and thereby increase the therapeutic potency of MAbs. Blood mononuclear cells (PBM) coated with CAMPATH 1G (pan lymphocyte reactive rat IgG2b) MAb were used as targets (T) and autologous lymphocytes or granulocytes were used as effector (E) cells. We studied function in normal donors as well as patients 1 week before and 3‐6 weeks after bone marrow transplantation (BMT). In the absence of CAMPATH 1G, specific 51Cr release from autologous PBM was < 1% in all groups, even after pre‐incubation of the effector cells with granulocyte‐macrophage. colony stimulating factor (GM‐CSF) or interleukin‐2 (IL2). In the presence of 5 μg/ml of MAb, and at an E: T ratio of 50:1, lymphocytes from normal donors induced a low level (6.5%) of 51Cr release from auto PBM rising to 9% after pre‐incubation of effector cells with IL2 (P= 0.02). Granulocytes had greater activity inducing 10%51CR release (range 2‐23%) which rose to 21.6% with GM‐CSF (range 12‐48%) (P0.001). Pre‐BMT, killing by lymphocytes and granulocytes was not significantly different from normal, and responded to IL2 and GM‐CSF. In contrast, granulocyte killing after BMT was significantly impaired (51Cr release 3%) and showed no rise with GM‐CSF. Killing by lymphocytes, however, remained normal, as did their IL2 response. Loss of granulocyte mediated MAb‐DCC coincided with significant post‐BMT impairment of oxidative metabolism: expression of Fc receptors II and III, however, was normal. Optimum therapeutic effect of MAb‐DCC is likely to be achieved when MAb are given together with appropriate cytokihes. the choice of which will depend upon the clinical circumstances.
Polyclonal rearrangements of the T‐cell receptor β‐chain in fatal angioimmunoblastic lymphadenopathy
Authors: Knecht Hans , Odermatt Bernhard F. , Hayoz Daniel , Kühn Lukas , Bachmann Fedor
Published: March 2008

Genomic rearrangement of germline T‐cell antigen receptor (TcR) and immunoglobulin (Ig) genes was studied by Southern blot analysis in seven patients with angioimmunoblastic lymphadenopathy (AILD). In three cases clinically suspected of transformation into malignant lymphoma, hybridization with the TcRβ probe showed markedly dimished intensity in the 11.5 kb germline band after Eco RI digestion and normal germline configuration after Hind III and Bam HI digestion, indicating polyclonal T cell rearrangements. A clonal rearrangement of the TcRβ gene was detected in only one case at initial biopsy. No monoclonal rearrangement of Ig genes was observed. These data show that in some cases of AILD disease progression is indicated by polyclonal TcR rearrangements and not by outgrowth of a malignant clone, supporting the concept of AILD as an immunoregulatory disorder.
Decreased stability and structural heterogeneity of the residual platelet glycoprotein IIb/IIIa complex in a variant of Glanzmann's thrombasthenia
Authors: Modderman P. W. , van Mourik J. A. , Berkel W. van , Cordell J. L. , Morel M. C. , Kaplan C. , Ouwehand W. H. , Huisman J. G. , von dem Borne A. E. G. Kr.
Published: March 2008

A patient is described with a disturbance of platelet function comparable to that in Glanzmann's thrombasthenia. Platelet aggregation and binding of fibrinogen to the patient's platelets were defective and thrombin‐induced clot retraction was absent. The platelet fibrinogen content was only moderately reduced. As measured by monoclonal antibody binding in the presence of divalent cations, the platelets contained about 15% of the normal amount of GPIIb and GPIIIa and only 6% of the normal amount of intact GPIIb/IIIa complex. The residual GPIIb/IIIa complex exhibited a decreased stability as shown by the lack of binding of a complex‐dependent anti‐GPIIb/IIIa antibody to platelets incubated with ethylene diamine tetraacetic acid (EDTA) at 22°C. Crossed immunoelectrophoresis (CIE) in the presence of divalent cations showed partial dissociation of GPIIb/IIIa as well as the presence of two forms of the residual intact GPIIb/IIIa complex. In addition, both CIE in the presence of the EDTA and two‐dimensional sodium dodecyl sulphate (SDS) gel electrophoresis showed the presence of two forms of GPIIb. This form of thrombasthenia is characterized by a defective platelet function, a marked reduction of GPIIb and GPIIIa, decreased stability of the residual GPIIb/IIIa complex and structural heterogeneity of GPIIb.
Blood from healthy animals and humans contains nondiscocytic erythrocytes
Authors: Simpson L. O.
Published: March 2008

Blood samples (three drops) from healthy animals and humans were fixed in a 2.5% solution of glutaraldehyde in 0.1 m cacodylate buffer at pH 7.4 within seconds of being drawn. Dehydrated and gold‐coated preparations were photographed at × 600 in a scanning electron microscope, then printed at × 1200. Erythrocytes were classified according to their shape and surface features into six classes, namely, normal red cells, flat cells, cells with surface changes, early cup forms, late cup forms, cells with altered margins. The size of each shape‐determined class was expressed as a percentage of the total number of cells counted. Data relating to 23 healthy females and to a mouse, rat, rabbit and horse are reported. Although the presence of nondiscocytic erythrocytes in peripheral human blood has been described, the presence of similar cells in animal blood has not been reported previously.
CD1 expression on B‐CLL lymphocytes
Authors: Merle‐Beral Héalégne , Boumsell Laurence , Michel Ariane , Debréa Patrice
Published: March 2008

Summary In a previous report we showed that D47 CD1a monoclonal antibody positively labelled B‐CLL cells of some patients. Six cases were selected to further characterize CD1 antigenic expression at the leukaemic cell surface using flow cytometry with a battery of six CD1a, two CD1b and two CD1c monoclonal antibodies. CLL cells were positively labelled with CD1a antibodies except OKT6 and NA1/34: in all cases they were CD1b negative and CD1c positive. These results suggested that CD1a, c epitopes can be detected on leukaemic B cells in addition to other T cell differentiation antigens. In view of recently published data demonstrating the presence of CD1c molecule in normal B cell subsets, our results further question the relationship of B‐CLL with a restricted subpopulation of B cells.
In vitro studies of ways to overcome resistance to VAMP‐high dose melphalan in the treatment of multiple myeloma
Authors: Millar B. C. , Bell J. B. G. , Maitland J. A. , Zuiable A. , Gore M. E. , Selby P. J. , McElwain T. J.
Published: March 2008

Summary Myeloma colonies (MY‐CFUc) from 7/24 patients undergoing treatment with VAMP (vincristine, adriamycin and methyl prednisolone) and high dose melphalan (HDM) were melphalan‐resistant. It was not possible to conclude that VAMP induced melphalan resistance in MY‐CFUc, but that resistance is endogenous in some myeloma cell populations. In 12/13 of the same patients of whom four had MY‐CFUc which were melphalan resistant, the sensitivity of MY‐CFUc and GM‐CFUc to busulphan was similar. Thus resistance of MY‐CFUc to melphalan did not confer resistance to busulphan.

MY‐CFUc from 1/7 of a second group of patients were adriamycin‐resistant. This resistance was removed when the cells were treated with a combination of verapamil (3 μg/ml) and adriamycin. Verapamil also enhanced the toxicity of adriamycin to MY‐CFUc, from two patients where there was no evidence for adriamycin resistance. In these three patients the sensitivity of both MY‐CFUc and GM‐CFUc was similar after treatment with verapamil.

Verapamil did not affect the uptake or efflux of 3H‐daunorubicin in sensitive and resistant RPMI‐8226 cells (myeloma) and peripheral blood mononuclear cells from a normal donor; neither did it affect the binding of 3H‐daunorubicin to nucleic acid.

It is concluded that verapamil may be a useful adjuvant to VAMP chemotherapy and that busulphan may provide an alternative to melphalan in patients whose myeloma cells are melphalan resistant.
Hairy cell leukaemia: observations on natural killer activity in different clinical stages of the disease
Authors: Demeter J. , Paloaczi K. , Lehoczky D. , Benczuar M.
Published: March 2008

Summary Follow‐up studies of natural killer (NK) cells, NK activity and antibody dependent cellular cytotoxicity (ADCC) in the course of hairy cell leukaemia (HCL) were carried out in a series of patients affected by the disease. NK activity against K562 targets was found to be high in all the patients with non‐symptomatic stable disease. On the other hand, absent or extremely low NK activity was found only in patients with symptomatic progressive disease. NK activity determined in the transitional stages showed values between these two extremes. Antibody dependent cellular cytotoxicity (ADCC) was less closely correlated with the clinical stage than NK activity. Our findings suggest that investigation of in vitro NK cell cytotoxicity might serve as a useful adjunct in determining clinical stage in HCL.
A recurrent missense mutation (Arg → Gln) and a partial deletion in factor VIII gene causing severe haemophilia A
Authors: Bernardi F. , Volinia S. , Patracchini P. , Gemmati D. , Boninsegna S. , Schwienbacher C. , Marchetti G.
Published: March 2008

Summary The presence of gene lesions in coagulation factor VIII (FVIII) gene was investigated in 70 Italian patients severely affected by haemophilia A. cDNA probes specific for exons 14–26 of the FVIII gene were used. In two related patients a gene deletion removes exon 26, a gene lesion similar to that described previously in a British haemophiliac. In exon 24 a C to T transition in the reverse complement strand causes a missense mutation in the coding strand (CGA → CAA, 2209 Arg → Gln). The mutation is located in a very conserved FVIII homology region and severely reduces FVIII activity. By restriction analysis and hybridizations with oligonucleotide probes this gene alteration was found in two unrelated haemophiliacs and in their relatives.
Late onset bone marrow failure associated with proximal fusion of radius and ulna: a new syndrome
Authors: Dokal I. , Ganly P. , Riebero I. , Marsh J. , Steed A. , Kendra J. , Drysdale C. , Hows J.
Published: March 2008

Summary Members of two unrelated families, one from England and the other from Portugal, presented with aplastic anaemia. Both families had clinical and laboratory features in common. The proposita presented with progressive bone marrow failure in adult life involving all three haemopoietic cell lines. In both families individuals with bone marrow failure also had proximal fusion of the radius and ulna bilaterally which segregates as an autosomal dominant trait. No cytogenetic defect was identified in myeloid or lymphoid cells, sampled from the marrow and peripheral blood.

Proximal fusion of the radius and ulna is uncommon and its co‐existance with late onset bone marrow failure may indicate an association. Growth failure, non‐skeletal congenital malformations and cytogenetic abnormalities were absent suggesting that this syndrome is distinct from Fanconi's anaemia. The onset in adult life of generalized bone marrow failure involving all three haemopoietic cell lines associated with proximal fusion of the radius and ulna also makes this syndrome distinct from the thrombocytopenia with absent radii (TAR) syndrome.
Unusual immunophenotypes in acute leukaemias: incidence and clinical correlations
Authors: Bradstock K. F. , Kirk J. , Grimsley P. G. , Kabral A. , Hughes W. G.
Published: March 2008

Summary The incidence and clinical implications of unusual patterns of expression of leucocyte differentiation antigens in acute leukaemia were assessed on 568 newly diagnosed paediatric and adult cases undergoing immunophenotyping with a panel of monoclonal antibodies at a single centre. Among patients with the precursor B (common) form of acute lymphoblastic leukaemia (ALL), the major variant seen was the group of 15 cases with expression of myeloid surface antigens. 4·5% of ALL cases tested with antibody to CD‐11b were positive, 5·1% were CD‐13+, and 10·8% CD‐33+. All 15 patients achieved a complete remission with chemotherapy, with six of eight children and four of seven adults remaining disease free. A smaller proportion (1–5%) of precursor B ALL patients showed expression of the T lineage marker, CD‐7. The only significant variant seen in the precursor T‐ALL group was expression of HLA‐DR antigen, which was found in five of 3 5 cases; although all responded to treatment, only one remains a disease‐free survivor. Among patients with acute myeloid leukaemia (AML), expression of the lymphoid markers terminal transferase (TdT) and CD‐7 were commonly seen (22·2% and 28·4% respectively of cases tested). Other lymphoid markers detected on AML cases were CD2 (11·1%), CD‐10 (1%) and CD‐19 (4·4%). These results confirm that examples of lineage infidelity are regularly seen in large series of patients with acute leukaemia. Prospective studies using uniform treatment protocols are required to establish whether these patients have significantly different disease outcomes.
Immunoglobulin heavy chain gene rearrangements in chronic lymphocytic leukaemia: correlation with clinical stage
Authors: Rechavi Gideon , Mandel Mathilda , Katzir Nurit , Brok‐Simoni Frida , Hakim Itzhak , Holtzman Fanny , Biniaminov Miriam , Givol David , Ben‐Bassat Isaac , Ramot Bracha
Published: March 2008

Summary A search for a correlation between the clinical stage of chronic lymphocytic leukaemia (CLL) and the pattern of immunoglobulin heavy chain gene rearrangements was undertaken. DNA samples from the leukaemic cells of 38 CLL patients were analysed by Southern blot hybridization. Using probes for the immunoglobulin heavy chain J (JH) and Cμ regions a marked heterogeneity of the hybridization patterns was observed in both regions. The number of JH hybridization bands varied from one to four and more than two were found in 58% of the patients. In 42% of the patients no germline JH genes were found. One to three additional Cμ bands were observed in 34%, but the germline was preserved in all samples. There was no correlation between the clinical stage and the number of hybridizing JH bands; however, a significant correlation was found between the loss of JH germline band or a Cμ multiband pattern and advanced stage of the disease. The genetic events in the immunoglobulin genes observed in advanced CLL patients are assumed to result from clonal evolution and tumour progression.
Complete recovery of marrow function after treatment with anti‐lymphocyte globulin is associated with high, whereas early failure and development of paroxysmal nocturnal haemoglobinuria are associated with low endogenous G‐CSA‐release
Authors: Nissen Catherine , Moser Yolanda , Carbonare Verena Dalle , Gratwohl Alois , Speck B.
Published: March 2008

Summary 24 patients who were treated with antilymphocyte globulin (ALG) for severe aplastic anaemia (SAA) were tested for endogenous release of granulocyte colony stimulating activity (G‐CSA) prior to, and at various intervals after treatment. CSA‐production in vitro was induced with auto‐ logous serum as a source of ‘releaser’ activity, avoiding the use of plant mitogens. Before treatment, G‐CSA‐release was highly variable. Though mean values were higher in the 17 patients who subsequently responded to ALG treatment than in the six non‐responders, this difference was not statistically significant. In the 17 responders, G‐CSA‐release strongly increased prior to improvement of peripheral blood counts. In one responder patient tested before, and at regular intervals after ALG, CSA‐release was high before, abnormally low at 7 d and increased again to high values before the onset of bone marrow reconstitution. In six patients who did not respond to ALG‐treatment, G‐CSA release decreased after treatment, and a second course of ALG was ineffective when given during this low CSA‐phase. Five of the 24 patients developed paroxysmal nocturnal haemoglobinuria (PNH) at 9 months to 3 years after ALG‐treatment. In all, the onset of PNH was associated with very low G‐CSA‐release, whether it had been high or low before treatment.

We conclude that low‐CSA‐release after ALG treatment is a poor prognostic sign. It either indicates progression of marrow failure or heralds PNH. Such patients may be candidates for early bone marrow transplantation or treatment with G‐CSF or GM‐CSF.
Lipid peroxidation of human granulocytes (PMN) and monocytes by iron complexes
Authors: Hoepelman I. M. , Bezemer W. A. , van Doornmalen E. , Verhoef J. , Marx J. J. M.
Published: March 2008

Summary Because the phagocytic function of non‐stimulated human polymorphonuclear granulocytes (PMN) is impaired after incubation with either polynuclear Fe(III) or Fe(II), we decided to study lipid peroxidation of PMN and monocytes by these iron complexes. Lipid peroxidation was assessed by measuring thiobarbituric acid reactive substances and fluorescent compounds. In this study we report that monocytes, isolated in the standard way (EDTA‐mediated detachment), release significantly more thiobarbituric acid reactive substances after incubation with iron than PMN. Monocytes, however, isolated without EDTA release the same amount after incubation with iron as PMN. The iron complexes shown to impair phagocyte function also stimulated the peroxidation of membrane lipids. Ascorbic acid in high concentrations enhanced iron‐induced lipid peroxidation. Lipid peroxidation induced by ferrous ascorbate (1:20) could be inhibited by catalase, the iron chelators deferoxamine and transferrin, and the hydroxyl radical scavenger thiourea. Mononuclear complexes (ferric citrate 1:20) did not impair granulocyte function and did not induce lipid peroxidation. Our results suggest that impaired phagocyte function in patients with iron overload may be due to non‐transferrin bound iron‐mediated peroxidation of membrane lipids. However, the lowest amount of ferrous ascorbate (1:20) capable of inducing lipid peroxidation (50 μm) was significantly higher than that which impaired phagocyte function (10 μm).
Chemosensitivity testing of fresh leukaemia cells using the MTT colorimetric assay
Authors: Twentyman Peter R. , Fox Norma E. , Rees John K. H.
Published: March 2008

We have determined the feasibility of using the MTT colorimetric assay for measuring the sensitivity of fresh leukaemia cells to cytotoxic agents. A linear relationship was seen between the number of leukaemic cells plated and the resulting absorbance in the MTT assay at day 4. For 2 × 105 cells/well from CLL peripheral lymphoctes, the mean absorbance (measured at 540 nm) was 0.41. Similar curves were obtained with the assay carried out between days 3 and 7. ID50 values for CLL lymphocytes exposed to adriamycin (ADM) were in the range of 0.02‐0.29 μg/ml, a similar range to that given by the DiSC dye exclusion assay. Sensitization of leukaemia cells to ADM by verpamil or cyclosporin A could be demonstrated in some patients but not in others. The lymphocytes from one patient with prolymphocytic leukaemia were abnormally resistant to ionizing radiation. These cells also showed resistance to melphalan and adriamycin but not to vincristine. The MTT assay appears to offer an attractive option for in vitro chemosensitivity testing in leukaemia, determination of cross‐resistance profiles and studies of resistance modifiers.
A translocation involving a specific breakpoint (q35) on chromosome 5 is characteristic of anaplastic large cell lymphoma (‘Ki‐1 lymphoma')
Authors: Rimokh Ruth , Magaud J.‐P. , Berger Francoise , Samarut J. , Coiffier B. , Germain D. , Mason D. Y.
Published: March 2008

Two cases of non‐Hodgkin's lymphoma are reported in which a chromosomal translocation was observed involving the same site (q35) on the long arm of chromosome 5. The other breakpoint involved in the translocation was chromosome 2 (p23) in one case and chromosome 3 (q12) in the other. Both cases were large cell lymphomas expressing CD30 antigen (‘Ki‐1 lymphoma'). One was clearly of T lymphoid origin, the other probably B cell derived. One other case of a Ki‐1 lymphoma with 2;5 translocation (involving the same breakpoint on chromosome 5) has been reported previously, and it is suggested that this cytogenetic abnormality may be specifically associated with Ki‐1 lymphoma. The literature contains a further eight cases of lymphoid neoplasms with a translocation involving a breakpoint at q35 on chromosome 5. They have all been described as cases of‘malignant histiocytosis’, but the present findings make it likely that these cases were in reality also examples of Ki‐1 lymphoma. The breakpoint at the q35 region on chromosome 5 is close to the position of the fms proto‐oncogene, suggesting that an abnormality affecting this gene might possibly play a causal role in ‘Ki‐1 lymphoma’. However, DNA restriction fragment analysis of the present cases showed no evidence that the breakpoint on chromosome 5 involves the fms gene or its immediate vicinity.
Mortality and causes of death in Dutch haemophiliacs, 1973–86
Authors: Rosendaal F. R. , Varekamp I. , Smit C. , Bröcker‐Vriends A. H. J. T. , Van Dijck H. , Vandenbroucke J. P. , Hermans J. , Suurmeijer T. P. B. M. , Briët E.
Published: March 2008

Mortality figures were calculated for a group of 717 Dutch haemophiliacs over the period 1973‐86. Follow‐up was on average 10.9 years; no patients were lost to follow‐up. The data were compared to the general male population by actuarial methods and patient‐year analysis. Forty‐three patients died, while 20 deaths were expected in a hypothetical group of non‐haemophiliacs of the same age distribution. Hence, overall mortality was 2.1 times higher than in the general population. This leads to a calculated life expectancy of 66 years, as compared to 74 years in the general male population. Mortality did not differ much by severity of haemophilia. A possibly beneficial effect of prophylaxis on longevity was observed. Haemorrhage occurred in half of all deaths and among these traumatic bleeding was the most prevalent. The number of deaths due to ischaemic heart disease was significantly lower (80% reduction) than expected and therefore the authors conclude that haemophilia offers protection against ischaemic heart disease. Cancer mortality was significantly higher (2.5 times) than expected.
Anti‐idiotypes to factor VIII antibodies and their possible role in the pathogenesis and treatment of factor VIII inhibitors
Authors: Moffat E. H. , Furlong R. A. , Dannatt A. H. G. , Bloom A. L. , Peake I. R.
Published: March 2008

Studies on the production and characterization of anti‐idiotype antibodies (AId) to monoclonal factor VIII antibodies (McFVIIIAb) are reported. Two AIds were produced and one of these exhibited cross‐reactivity with two other McFVIIIAb but showed no reactivity with haemopholic and non‐haemophilic FVIIIAb. This AId was also active against McFVIIIAb which bound immunologically active forms of factor VIII but it did not neutralize McFVIIIAb directed against procoagulant factor VIII. The in vitro effect of therapeutic pooled human IgG concentrates upon haemophilic and non‐haemophilic FVIIIAb was also assessed. Approximately 25% of non‐haemophilic FVIIIAb were inhibited following incubation with human IgG whereas the latter had no effect upon haemophilic FVIIIAb activity. Studies on haemophilic sibling pairs of FVIIIAb and non‐FVIIIAb producing individuals indicated FVIIIAb neutralizing activity in the non‐FVIIIAb producing sibling's IgG fraction which may be of anti‐idiotypic origin. These findings lend further support to suggestions that anti‐idiotypes have a regulatory role in FVIIIAb production and are of potential therapeutic value.
Severe Hb S‐β°‐thalassaemia with a T → C substitution in the donor splice site of the first intron of the β‐globin gene
Authors: Gonzalez‐Redondo J. M. , Stoming T. A. , Kutlar F. , Kutlar A. , McKie V. C. , McKie K. M. , Huisman T. H. J.
Published: March 2008

Through direct sequencing and dot‐blot analyses with synthetic oligonucleotide probes of amplified DNA, a new nucleotide substitution was discovered in a Black teenager with severe Hb‐S‐β°‐thalassaemia. The substitution involved a T → C replacement at the second position of the donor splice site of the first intervening sequence of the β‐globin gene. The clinical and haematological observations made in Black subjects with Hb S‐β°‐thalassaemia, with different types of thalassaemia, suggest severe disease resembling sickle cell anaemia. Only an occasional patient had a milder clinical course, perhaps because of a greatly increased production of fetal haemoglobin.
In vivo regeneration of red cell 2, 3‐diphosphoglycerate following transfusion of DPG‐depleted AS‐1, AS‐3 and CPDA‐1 red cells
Authors: Heaton Andrew , Keegan Thais , Holme Stein
Published: March 2008

Regeneration of 2, 3‐diphosphoglycerate (DPG) was determined following transfusion of DPG‐depleted group O red cells into group A recipients. Blood from five donors was stored in the adenine‐containing solutions CPDA‐1, AS‐1 or AS‐3 for 35 d at 4°C. Post‐transfusion red cell DPG and ATP were measured in separated group O red cells over a 7 d period. The studies confirmed rapid in vivo DPG regeneration with ≥ 50% of the maximum level being achieved within 7 h. An average of 95% of the recipients’ pre‐transfusion DPG level was achieved by 72 h and by 7 d mean (±SEM) DPG levels relative to recipient's pre‐transfusion DPG averaged 84% (± 13%), 92% (± 17%) and 84% (± 21%) for CPDA‐1, AS‐1 and AS‐3 red cells, respectively. Results were comparable to those previously reported for blood stored in ACD for 15‐20 d (Valeri & Hirsch, 1969; Beutler & Wood, 1969). The immediate regeneration rate, V, closely approximated first order regeneration kinetics with AS‐3 red cells exhibiting double the rate of CPDA‐1 red cells (P<0·001). AS‐1 red cells exhibited an intermediate rate of regeneration which was not significantly different compared to either CPDA‐1 or AS‐3 (P>0·05). V exhibited a significant (P<0·05) positive correlation with ATP levels 5‐7 h post‐infusion. ATP regeneration of the infused cells was rapid with a mean increase of 1·2 μmol/g Hb above post‐storage levels being achieved 1 h following transfusion.
Effect of cell growth and cell differentiation on 1‐β‐d‐arabinofuranosylcytosine metabolism in myeloid cells
Authors: Chiba P. , Tihan T. , Eher R. , Köller U. , Wallner C. , Göbl R. , Linkesch W.
Published: March 2008

Summary. Resistance of leukaemic blasts to 1‐β‐D‐arabinofuranosylcytosine (ara‐C) has been shown to be associated with changes in the metabolism of this drug. However, effects of cell growth and maturation stage on ara‐C metabolizing enzymes have to be excluded as a possible cause of different enzyme activities in leukaemic blasts between nonresponders and patients achieving complete remission. We evaluated the effects of cell cycle phase and cell differentiation on the activity of cytidine deaminase, deoxycytidylate deaminase and deoxycytidine kinase in myeloid cell lines. Our data indicate that different enzyme profiles in nonresponders might not only be caused by the emergence of mutator phenotypes but may also reflect the growth and maturation stage of leukaemic blasts.
Identification of normal B‐cell counterparts of neoplastic cells which secrete cold agglutinins of anti‐I and anti‐i specificity
Authors: Stevenson F. K. , J. North G. J. Smith , Hamblin T. J. , Glennie M. J.
Published: March 2008

Summary. A monoclonal antibody, which recognizes a cross‐reacting idiotypic determinant present on human cold‐reactive autoantibodies with anti‐I or anti‐i binding activity, has been found to specifically inhibit the cold agglutination of red cells. This suggests that an epitope close to the binding site of such autoantibodies is being recognized.

The antibody has been used to identify tumour cells in the blood of three patients with cold haemagglutinin disease, and to analyse the heterogeneous nature of the neoplastic B‐cell clone present in the bone marrow of one of the patients. Using S‐phase analysis, it was found that cell proliferation was occurring in the bone marrow but not in the blood, and that the major proliferating population was that of lymphoplasmacytoid cells containing large vesicular inclusions of idiotypic IgM.

It has also been possible to locate normal B‐cells which are recognized by the anti‐idiotypic antibody. Such cells have been found throughout the normal adult lymphoid tissue where they account for 2.9–10.8% of the B lymphocyte population. They are also present in fetal spleen at 15 weeks gestation, indicating that immunoglobulins bearing this sequence form part of the immature B‐cell repertoire.
Clinical stage of chronic granulocytic leukaemia and BCR breakpoint location in South‐East Asian patients
Authors: Dyck Jacqueline A. , Bosco John J.
Published: March 2008

Summary. Forty‐six Malaysian patients with chronic granulocytic leukaemia were found to be rearranged in the breakpoint cluster region (BCR) of chromosome 22, molecular evidence of Philadelphia chromosome (t9,22) translocation. Through the use of a 1–2 kb 3’BCR probe and two restriction enzyme digests, patients’breakpoints could be localized either to 5’or 3’regions of the BCR. Breakpoint site localization at the time of DNA sampling did not show any positive statistical association to clinical status defined as chronic phase, chronic phase with less than 6 months to blast crisis, accelerated phase and blast crisis. This was in contrast to earlier reports which indicated that patients with breakpoint at 3′ site were at a higher biologic risk for entering blast crisis.
Dyserythropoiesis and ineffective erythropoiesis in Plasmodium vivax malaria
Authors: Wickramasinghe S. N. , Looareesuwan Sornchai , Nagachinta Bussarin , White N. J.
Published: March 2008

Summary. Nine Thai adults with P. vivax malaria were investigated. Light and electron microscope studies of marrow aspirates revealed morphological evidence of dyserythro‐poiesis in six of them, Dyserythropoiesis was most marked in the four most anaemic patients. In these four patients the electron microscope also revealed the presence of erythro‐blasts at various stages of degradation within the cytoplasm of macrophages. Neither the dyserythropoiesis nor the ineffective erythropoiesis could be attributed to a deficiency of vitamin B12, folate or iron. The abnormalities of erythropoiesis seemed to result from the P. vivax infection itself. Other bone marrow reactions seen in this infection included macrophage hyperplasia, plasmacytosis and increased eosinophil granulocytopoiesis. Unlike in severe P. falciparum malaria, the microvasculature of the marrow was not obstructed by parasitized red cells.
Prognostic factors in chronic lymphocytic leukaemia: the importance of age, sex and response to treatment in survival
Authors: Catovsky D. , Fooks J. , Richards S.
Published: March 2008

Summary.We report the analysis of prognostic factors in a cohort of 660 patients entered in the first Medical Research Council trial in chronic lymphocytic leukaemia (CLL) between 1978 and 1984. The majority (94%) of patients were aged 50 or over and the number of men (M) was almost twice that of women (F) with an M: F ratio of 1·8: 1. The M: F ratio was lower, 1·5:1, in patients aged 70 or over. Stage A CLL was the most common, and stage C the least common, among women of all ages, in contrast to men for whom stage A only predominated in the older age group. As the majority of CLL patients are elderly we have examined the causes of death in great detail. 29% of deaths were unrelated to CLL, mainly other cancers (12%) and cardiovascular complications (16%). The majority of deaths in patients presenting with stages B and C were from CLL‐related causes, whilst almost half of the deaths in patients presenting with stage A were not obviously related to CLL. Univariate analysis disclosed that the A, B, C staging system was the most important factor considered; stratified and multivariate analysis showed that age and response to treatment were the main prognostic factors after stage. Women always fared better than men and this was independent of stage and age. This and other features documented in the trial suggest a major biological difference between the sexes which has not been widely recognized. The significant influence of treatment response on patients’ survival suggests that the search for better treatments in CLL may be rewarding. The improved median survival of stage C patients recorded in this trial, 41 months, compares favourably with previous reports and may have resulted from better treatment.
Surface marker expression in adult acute myeloid leukaemia: correlations with initial characteristics, morphology and response to therapy
Authors: Campos Lydia , Guyotat Denis , Archimbaud Eric , Devaux Yves , Treille Danielle , Larese Alain , Maupas Jean , Gentilhomme Odile , Ehrsam Alain , Fiere Denis
Published: March 2008

Summary.The clinical significance of surface markers was investigated in 145 cases of acute myeloid (AML) or undifferentiated leukaemia (AUL), using a panel of six monoclonal antibodies directed to NHL‐30.5 antigen (expressed on poorly differentiated myeloid cells), CD13, CD14, CD15, CD33 and CD34 antigens. Expression of CD14 was correlated with higher leucocyte count, higher serum lactate dehydrogenase level and presentation with extramedullary disease. There was no strict correlation with the French‐American‐British classification. However, the expression of CD14 was associated with monocytic subtypes. CD15 was mainly expressed in M2 and M3 subtypes, and NHL‐30.5 and CD34 antigens in AUL and M1 leukaemias. All patients were treated with the same intensive induction treatment. Staining by three antibodies had a prognostic value. The complete remission (CR) rates were 38% (26/68) in NHL‐30.5‐positive versus 75% (62/77) in NHL‐30.5‐negativecases(P<10−5), 50%(37/74) in CD34‐positive versus 72% (51/71) in CD34‐negativecases (P= 0.007) and 70% (77/110) in CD15‐positive versus 31% (11/35) in CD15‐negative cases (P<10−4). Expression of NHL‐30.5 and CD34 antigen was associated with shorter survival (P< 10−3andP< 10−2respectively), whereas survival was longer in CD15‐positive cases (P< 10−3). In multi‐ variate analysis, expression of NHL‐30.5 antigen, absence of CD15, and high LDH level were associated with poor survival. CR duration was not influenced by any of the factors studied, including antigen expression. These results suggest that leukaemias with less differentiated phenotype have a lower response rate to induction treatment.
Interferon alpha‐2b as treatment for Philadelphia‐negative chronic myeloproliferative disorders with excessive thrombocytosis
Authors: Lazzarino M. , Vitale A. , Morra E. , Gagliardi A. , Bernasconi P. , Torromeo C. , Inverardi D. , Burgio V. , Castello A. , Bernasconi C. , Mandelli F.
Published: March 2008

Summary. We treated 32 patients with Ph1‐negative chronic myeloproliferative disorders (CMD) with excessive thrombocytosis with Interferon α‐2b (IFN α‐2b): 26 had essential thrombocythaemia, ET (18 previously untreated, eight pretreated); one thrombocythaemia after treatment for Hodgkin's disease (HD); two thrombocythaemia associated with non‐Hodgkin's lymphoma (NHL); three stage II idiopathic myelofibrosis (IM). IFN was given at daily doses of 1−4 × 106 IU. Twenty‐seven patients (84%) responded, 17 (53%) achieved complete haematologic response after a median time of 12 weeks, and 10 (31%) partial haematologic response. Median platelet levels declined in complete haematologic response patients from 1190 to 335 × 109/I. Normalization of megakaryocyte (MK) levels was observed in 8/17 complete haematologic response patients treated for 9–12 months, with decreased bone marrow (BM) cellularity. Side effects requiring dose reduction or discontinuation of treatment occurred in 28% of cases with IFN doses of 2 or 4 × 106 IU. After 1 year of continuous IFN treatment, responses were maintained with conventional chemotherapy or low‐dose IFN. This study demonstrates that IFN has definite therapeutic activity in CMD with excessive thrombocytosis. This biological agent, either alone or in combination with other antineoplastic treatment, may represent a new therapeutic approach for these disorders.
18q21 rearrangement in diffuse large cell lymphoma: incidence and clinical significance
Authors: Offit Kenneth , Koduru Prasad R. K. , Hollis Rachel , Filippa Daniel , Jhanwar Suresh C. , Clarkson Bayard C. , Chaganti R. S. K.
Published: March 2008

Summary. Cytogenetic, molecular genetic and clinical information was collected for 102 cases of diffuse large cell lymphoma (DLCL) ascertained in a diagnostic laboratory over a 3‐year period. Nineteen cases showed evidence of either a t(14;18) or a rearrangement of one of three genomic probes for breakpoints at 18q21. Clinical and histologic evidence of transformation from follicular lymphoma or chronic lymphocytic leukaemia was available in six cases. Except for age, prognostic clinical variables (LDH, stage, extranodal involvement) were similar between the 18q21 rearranged patients and DLCL patients without 18q21 rearrangement. At a median follow‐up in excess of 2 years for both groups, there was no difference in overall survival between the 18q21 rearranged group compared to DLCL patients lacking this genetic abnormality. The median disease‐free survival for the 18q21 rearranged group, however, was significantly shorter and survival in partial remission longer. The propensity for extended survival of the 18q21 rearranged DLCL patients with residual or recurrent disease resembled the clinical behaviour of nodular lymphoma patients with t(14;18). These results suggest that cytogenic or molecular genetic identification of a chromosome 18q21 translocation may be of prognostic significance in the analysis of treatment protocols for patients with DLCL.
Defective propeptide processing and abnormal activation underlie the molecular pathology of factor IX Troed‐y‐Rhiw
Authors: Liddell M. B. , Lillicrap D. P. , Peake I. R. , Bloom A. L.
Published: March 2008

Summary. Affected members of a South Wales haemophilia B family (from Troed‐y‐Rhiw) were shown by Western blotting and immunoperoxidase detection to have a factor IX molecule of higher than normal molecular weight which also shows impaired calcium binding. Gene cloning and DNA sequencing revealed the same arginine to glutamine mutation at position ‐ 4 of the propeptide that has been found in two previously described factor IX variants which circulate with the propeptide still attached. The mutation also abolishes a HaeIII restriction enzyme recognition site. A potential carrier was shown to be normal both by Western blotting and DNA studies. The way in which the attached propeptide interferes with normal factor IX function was investigated by activation studies with crude normal and patient factor IX Troed‐y‐Rhiw preparations using Western blotting and detection with iodinated immunopurified polyclonal anti‐factor IX serum. We demonstrate that the −4 mutation appears to block cleavage between the Arg 145‐Ala146 peptide bond in the activation peptide, thus preventing the normal activation of factor IX Troed‐y‐Rhiw. A small amount of normally processed factor IX is produced, implying that the −4 mutation does not completely prevent propeptide cleavage, thus accounting for the low levels of factor IX activity measured in the plasma of affected family members.
Factor IX Kawachinagano: impaired function of the Gla‐domain caused by attached propeptide region due to substitution of arginine by glutamine at position −4
Authors: Sugimoto M. , Miyata T. , Kawabata S. , Yoshioka A. , Fukui H. , Iwanaga S.
Published: March 2008

Summary. Factor IX Kawachinagano (KWC) is a mutant factor IX protein initially recognized in a patient with severe haemophilia B, who had 46% of normal factor IX antigen and no detectable clotting activity. Previous studies indicated that factor IX KWC was not activated by factor XIa in the presence of Ca ions. In the present study, we purified and analysed factor IX KWC at a structural level in an attempt to clarify the nature of the impaired reaction with factor XIa. Kinetic studies showed that activation of factor IX KWC by factor X activator from Russell's viper venom (RVV‐X) was normal, whereas activation by factor XIa was defective. Amino acid sequence analysis of tryptic peptides and direct analysis of the NH2‐terminal sequence of factor IX KWC demonstrated that this mutant factor IX retained the propeptide region of 18 amino acids due to a substitution of arginine ‐(‐4) by glutamine. These data suggested that the Gla‐domain of factor IX KWC was dysfunctional, although the total γ‐Gla content, measured by alkaline‐hydrolysis, was normal. We assumed that this attached propeptide region of the molecule directly interferes with the adjacent NH2‐terminus and prevents the metal‐induced conformational changes which are essential for biological activity of normal factor IX.
Congenital severe protein C deficiency in adults
Authors: Melissari E. , Kakkar V. V.
Published: March 2008

Summary. Two unrelated families are described, of which four adult members were found to be suffering from severe protein C deficiency (≤ 5% of normal plasma level). Newborn deaths were reported in the first family but the second family had no history of neonatal purpura fulminans and infant death. Thrombotic symptoms developed mainly in their early twenties, consisting chiefly of recurrent superficial and deep iliofemoral vein thromboses and pulmonary emboli. Other clinical features included generalized peritonitis due to massive mesenteric vein thrombosis, thrombosis of the cavernus sinus, renal vein thrombosis and priapism. In the second family, five members (all aged approximately 40 years) died of intravascular abdominal thrombosis. Massive thromboembolic episodes were associated with a compensated disseminated intravascular coagulation syndrome as evidenced by high concentrations of D‐dimer (mean levels 5000 ng/ml plasma) and by a moderate reduction in platelet count and fibrinogen concentrations. D‐dimer levels in non‐crisis periods were also raised above normal (mean levels 400 ng/ml). The thrombotic problems of these patients were controlled satisfactorily by long‐term administration of low molecular weight heparin alone or in combination with low dose warfarin.
Interferon therapy for chronic non‐A non‐B and chronic delta liver disease in haemophilia
Authors: Lee Christine A. , Kernoff P. B. A. , Karayiannis P. , Thomas H. C.
Published: March 2008

Summary. The case histories of a carrier of haemophilia A with chronic post transfusion non‐A non‐B hepatitis and a severe haemophiliac with chronic delta hepatitis are described. Therapy with lymphoblastoid alpha interferon resulted in improvement of NANB and HDV related chronic hepatitis and resolution of HIV related thrombocytopenia. Interferon may modulate replication of more than one transfusion transmitted virus in the haemophiliac.