bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of phenotypic assessment and the use of two restriction fragment length polymorphisms in the diagnosis of the carrier state in haemophilia B
Authors: Lillicrap D. P. , Liddell M. B. , Matthews R. J. , Peake I. R. , Bloom A. L.
Published: March 2008

Summary. Carrier detection in haemophilia B has previously involved pedigree analysis and assessment of the coagulation phenotype. At best using these methods, carrier evaluation may be made with about 80% certainty (Orstavik et al, 1981). With isolation and cloning of the factor IX gene, DNA probes are now available to detect intragenic nucleotide changes. This study uses one such probe which detects restriction fragment length polymorphisms with the restriction endonucleases Taq 1 and Xmn 1. Seventy‐eight individuals from eight haemophilia B kindred were initially evaluated for factor IXC and factor IXAg levels. DNA from these individuals was then digested with the two restriction enzymes and after Southern blotting, analysed for restriction fragment length polymorphisms (RFLPs) with the radiolabelled genomic probe. Four kindred proved informative with RFLP linkage assessment. In three families evaluation was possible with both Taq] and Xmn 1 polymorphic markers but in the fourth pedigree only Xmn 1 was informative. Using these techniques five potential carriers have been definitively assigned as either normal or carriers of the haemophilia B gene defect. Problems of phenotypic assessment are well illustrated in pedigree 4 where polymorphic linkage positively identified a carrier whose coagulation data were normal.
A study of lymphocyte subsets in patients with angioimmunoblastic lymphadenopathy
Authors: Pálóczi K. , Surányi P. , Nemes Z. , Szegedi Gy.
Published: March 2008

Summary Immunological marker studies of peripheral blood lymphocytes in 22 patients with typical angioimmunoblastic lymphadenopathy were carried out. There was a decrease of the absolute lymphocyte number, of T‐helper cells and the T4/T8 ratio in the active stage of the disease. The active T‐cells and total Tcells were reduced both in the active and in the inactive state. The number of B‐cells was in both states increased. Implications and possible mechanism of these observations are discussed.
Human granulocytes lack red cell Kx antigen
Authors: Branch Donald R. , Gaidulis Laima , Lazar Gary S.
Published: March 2008

Summary We have investigated the presence or absence of the red cell Kx antigen on human granulocytes by measuring specific uptake of anti‐Kx using three techniques: direct measurement by 125I‐staphylococcal protein A (125ISPA) and avidin‐biotin‐complex (ABC) immunoperoxidase staining and also, an indirect measurement using granulocyte adsorption of anti‐Kx, Our results with all three methods indicate that the Kx antigen is not present on normal human granulocytes. Prior to adsorption of the anti‐Kx serum with purified, pooled, normal human granulocytes, 11 of 21 (53%) of normal granulocytes were non‐reactive by 125I‐SPA and 16 of 20 (80%) by ABC. This pattern of reactivity was shown to be due to contamination of our anti‐Kx serum with an antibody to a granulocyte‐specific antigen unrelated to the Kx antigen. After adsorption, there was no diminution in the reactivity of the adsorbed anti‐Kx compared to the unadsorbed antiserum against red cells which express strong Kx antigen, i.e. Ko and DTT‐modified normal human red cells, by either serologic or 125I‐SPA techniques. Likewise, reactivity with McLeod red cells, which have weak expression of the Kx antigen, was not changed using either the unadsorbed or adsorbed anti‐Kx. The adsorbed anti‐Kx was nonreactive with all 12 normal donors’granulocytes tested by 125I‐SPA and with 10 normal donors’granulocytes tested by ABC. Furthermore, granulocytes from a Ko individual were nonreactive using either unadsorbed or adsorbed anti‐Kx. These studies indicate that Kx antigen is not present on normal human granulocytes. Further, additional adsorption studies using granulocytes from a boy with X‐linked chronic granulomatous disease (CGD) indicated that these granulocytes also do not possess the Kx antigen. In contrast to previous reports, these data suggest that Kx antigen is most probably a red cell‐specific antigen and that the red cell Kx antigen has no direct relationship to the biochemical defect in CGD.
Determination of the plasma cell labelling index with bromodeoxyuridine in a double fluorescence technique
Authors: Lokhorst Henk M. , Boom Saskia E. , Bast Bert J. E. G. , Ballieux Rudy E.
Published: March 2008

Summary. A new technique is described in which plasma cells actively synthesizing DNA can be identified in a heterogeneous cell population such as bone marrow. This method uses bromodeoxyuridine (BrdU) and a fluorescent monoclonal antibody against BrdU in combination with cytoplasmic staining for immunoglobulin (Ig). In 26 patients with monoclonal gammopathy (MG) the plasma cell labelling index (LI) as determined by this immunofluorescent method was compared with the tritiated thymidine (3H‐TdR) LI. No difference in sensitivity was found between the two methods. As the determination of the plasma cell LI with the BrdU/anti BrdU method is easy to perform and results can be obtained within 4 h, this immunofluorescent method seems to be an attractive alternative to the laborious time‐consuming classic 3H‐TdR LI.
Alloantibody‐induced platelet serotonin release is blocked by antibody to the platelet PL antigen
Authors: Duncan J. R. , Rosse W. F.
Published: March 2008

Summary. Anti‐platelet antibodies are known to activate the platelet release mechanism, usually measured by [14C]serotonin secretion. We have made immunoglobulin preparations of antibodies to HLA antigens, blood group A and platelet‐specific P1A1 antigen on the platelet surface and have defined the optimal conditions for [14C]serotonin secretion by washed platelets. For optimal [14C]serotonin release, platelets were washed in apyrase and suspended in Tyrode‐albumin with Ca++ and Mg++ and fibrinogen. Anti‐P1A1 at high concentration did not cause [14C]serotonin release and inhibited serotonin release by other alloantibodies to antigens on the platelet surface. This was shown to be a result of the inhibition of fibrinogen binding to the platelet by bound anti‐P1A1.
Agranulocytosis induced by propylthiouracil: evidence of a drug dependent antibody reacting with granulocytes, monocytes and haematopoietic progenitor cells
Authors: Fibbe Willem E. , Claas Frans H. J. , Van der Star‐Dijkstra Wies , Schaafsma M. Ronald , Meyboom Ronald H. B. , Falkenburg J. H. Frederik
Published: March 2008

Summary. A patient with agranulocytosis and myeloid marrow hypoplasia following a second exposure to propylthiouracil (PTU) was studied for antibodies against mature blood cells and bone marrow precursor cells. During the acute phase of the agranulocytosis, significant growth inhibition of the myeloid committed progenitor cells (CFU‐GM) was found following incubation with complement, indicating the presence of in‐vivo cell bound cytotoxic antibodies. Using immunofluorescence and complement dependent cytotoxicity techniques it was demonstrated that acute phase and recovery phase sera contained circulating antibodies, reactive not only with differentiated granulocytes and monocytes but also with myeloid and erythroid (BFU‐E/CFU‐E) progenitor cells. Complement dependent lysis of the progenitor cells was facilitated by preincubation with PTU. These results indicate that the agranulocytosis was mediated by a PTU dependent antibody that affected both mature blood cells and bone precursor cells.
Increased frequency of DR2 in patients with aplastic anaemia and increased DR sharing in their parents
Authors: Chapuis B. , Von Fliedner V. E. , Jeannet M. , Merica H. , Vuagnat P. , Gratwohl A. , Nissen C. , Speck B.
Published: March 2008

Summary

68 patients with aplastic anaemia, their parents and healthy siblings were typed for HLA‐A, B and DR antigens. Among the patients there is an overrepresentation of DR2. The parents of affected children show an increased compatibility at the DR locus. This situation could favour the expression of recessive gene(s) involved in haematopoiesis and located in the HLA locus.
G6PD‐deficiency infectious haemolysis: a complement dependent innocent bystander phenomenon
Authors: Kasper Michael L. , Miller Wesley J. , Jacob Harry S.
Published: March 2008

Summary

Dramatic haemolysis may accompany viral hepatitis and pneumococcal pneumonia in G6PD‐deficient patients. Since red blood cells (RBCs) are richly endowed with receptors for activated complement, particularly C3b, we hypothesized that bulky, complement‐activating immune complexes (IC) consisting of microbes and antibody might attract granulocytes (PMNs), facilitating oxidative‘innocent bystander’RBC damage. Indeed, opsonization with only two type‐2 pneumococcus (PN3)/anti‐PN3/C3b complexes per RBC caused agglutination of RBC, a phenomenon termed immune adherence. Addition of as few as one PMN per 20 opsonized RBCs caused the glutathione (GSH) levels of co‐incubated G6PD‐deficient RBCs to fall by 30% (from 3.5 to 1.8 ± 0.8 μmoles GSH/g Hb) compared to identically incubated, but non‐opsonized, G6PD‐deficient RBCs. GSH levels remained normal (5.2 ± 0.4 μmoles/g Hb) in PMN‐exposed opsonized normal RBCs. GSH depletion in G6PD‐deficient RBC was directly related to disease severity—falling a mean 33% in RBCs from two Black G6PD A‐ subjects but 59% in two Caucasian G6PD deficient RBCs. Prevention of C3b generation (with 10 mm EDTA) during opsonization abrogated both immune adherence and PMN‐mediated GSH decline in oxidant‐sensitive cells. Similarly, removal of C3b receptors by brief trypsin incubation of RBCs eliminated immune adherence and GSH decline. Thus, both phenomena are dependent on IC complement activation and subsequent binding of the bacterial IC to the RBC complement receptors. Although clearance of IC by RBCs may be beneficial in protecting other tissues from inflammatory damage, G6PD‐deficient RBCs are vulnerable to oxidants generated by juxtaposed phagocytes—cells attracted to, and stimulated by, the immune complex/C3b combination. It is suggested that this‘Good Samaritan’activity of RBCs may lead to haemolysis during periods of exuberant antibody response to microbes.
Biochemical characterization of human myeloperoxidase using three specific monoclonal antibodies
Authors: Morishita Yoshihisa , Morishima Yasuo , Ogura Mlchinori , Nagai Yoshiyuki , Ohno Ryuzo
Published: March 2008

Summary

We have developed three monoclonal antibodies (moAbs), MA1, MA3 and MB1, which react with different antigenic determinants of human myeloperoxidase (MPO). In MPO‐positive culture cell lines, HL‐60 and NKM1, analysis of MPO by pulse‐chase experiments followed by immunoprecipitation with these moAbs revealed that MPO was composed of subunits of 59K, 18K and 14.8K dalton polypeptides which are plausibly derived from the 89 K precursor. MA1 and MB1 react with both the precursor and the mature forms of MPO. MA3 reacts with only the mature forms of MPO. Blocking experiments on MPO‐related functions revealed that the three moAbs could be divided into two groups. MA1 and MA3 inhibit MPO activities such as tetraguaiacol formation, iodide oxidation and luminol‐dependent chemiluminescence, while MB1 shows no such inhibition.
Infective complications of aplastic anaemia
Authors: Keidan A. J. , Tsatalas C. , Cohen J. , Cousins S. , Gordon‐Smith E. C.
Published: March 2008

Summary

Patients with aplastic anaemia have a relatively specific immune defect—leucopenia with neutropenia. We have carried out a retrospective analysis of infective episodes in 11 patients with aplastic anaemia. 5723 follow‐up days accrued and 29 infective episodes were documented. Overall the number of infective episodes was significantly associated with the mean white cell and monocyte counts (r = 0.9, 0.02<P<0.05) but not with length of follow‐up, presentation values of white cell, neutrophil and monocyte counts, or mean neutrophil count. The patients appeared to divide clinically into two groups, those at low risk (seven patients) and those high risk (four patients) of infection. Patients in the high risk group had significantly more infections (P = 0.01) and significantly lower monocyte counts (0.02 <P<0.05) than patients in the low risk group. These results are in contrast to similar studies in patients with chemotherapy‐induced neutropenia; in our patients the overall rate and severity of infection was low, Gram negative infections were uncommon and monocytopenia appeared to be of greater importance than neutropenia in determining susceptibility to infection.
Platelet‐derived growth factor stimulates growth of highly enriched multipotent haemopoietic progenitors
Authors: Michalevicz Rita , Katz Fay , Stroobant Paul , Janossy George , Tindle Robert W. , Hoffbrand A. Victor
Published: March 2008

Summary

Platelet‐derived growth factor (PDGF) has been shown to stimulate growth of normal and malignant fibroblasts, glial cells and smooth muscle cells. A growth promoting effect on human haemopoietic precursors has also been described, but the interpretation of this haemopoietic proliferative response to PDGF has been hampered by the lack of purity of the target population. In this study we show that PDGF promotes growth of early bone marrow haemopoietic progenitors depleted of either monocytes or T lymphocytes which are known to influence haemopoiesis. Moreover, the action of PDGF is even increased on a highly enriched BI‐3C5 early bone marrow population. BI‐3C5 is a novel monoclonal antibody which recognizes an antigen present on all multilineage colony‐forming cells (CFU‐mix) (Tindle et al, 1985). BI‐3C5 positively and negatively sorted fractions were obtained by fluorescence activated cell sorting (FACS) and PDGF was found to stimulate growth of CFU‐mix in the BI‐3C5‐positive fraction (consisting of only 4–6% of the marrow population), the effect being more marked than that on unsorted bone marrow. The results suggest that the product of the cellular proto‐oncogene c‐sis (the putative structural gene for the β chain of PDGF) may play a regulatory role in the in vivo proliferation of multipotent haemopoietic progenitors.
Prediction of response of patients with acute nonlymphocytic leukaemia to remission induction therapy: use of clinical measurements
Authors: Preisler H. D. , Priore R. , Azarnia N. , Barcos M. , Raza A. , Rakowski I. , Vogler R. , Wlnton E. L. , Browman G. , Goldberg J. , Gottlieb A. J. , Grunwald H. , Rai K. , Mlller K. , Brennan J. , Chervenick P. , Joyce R. , Tricot G.
Published: March 2008

Summary

Two hundred patients with acute nonlymphocytic leukaemia received remission induction therapy consisting of cytosine arabinoside and an anthracycline antibiotic. Analysis of the pretherapy characteristics of the patients demonstrated that patient age was the most important factor in determining whether or not the patient would survive remission induction therapy. Assessment of the characteristics of the bone marrow after 6 d of therapy permitted the recognition of patients who were likely to fail to enter remission because of persistent leukaemia. Taken together, these observations demonstrate that it is possible to identify patients for whom conventional chemotherapy is not likely to be of benefit either because it is too intensive or because it is not intensive enough to produce a complete remission.
Production of anti‐cytomegalovirus antibody following T‐cell depleted bone marrow transplant
Authors: Wimperis J. Z. , Berry N. J. , Brenner M. K. , Grundy J. , Hoffbrand A. V. , Griffiths P. D. , Prentice H. G.
Published: March 2008

Summary

The serological status to cytomegalovirus (CMV) was examined in 24 patients with no detectable CMV excretion, following T‐iymphocyte depleted bone marrow transplantation. The results show that seropositive recipients continue to produce CMV antibody regardless of the serological status of the donor but that seronegative recipients fail to produce CMV antibody even when the donor is seropositive. These findings suggest that when an effective CMV vaccine becomes available vaccination of the donor would be unlikely to confer protection on the recipient but that vaccination of the recipient may achieve this.
Diagnostic significance of detecting pseudo‐Pelger‐Huet anomalies and micro‐megakaryocytes in myelodysplastic syndrome
Authors: Kuriyama K. , Tomonaga M. , Matsuo T. , Glnnai I. , Ichimaru M.
Published: March 2008

Summary

To investigate the kind of morphological dysplastic changes important in the diagnosis of myelodysplastic syndrome (MDS), we examined 50 patients with MDS and 86 control subjects. Micromegakaryocytes and pseudo‐Pelger‐Huët anomalies were detected in 46 patients (92%) with MDS, and were also found in the differentiated types of acute myeloid leukaemia and chronic myeloid leukaemia, but not in other haematological disorders, or healthy subjects. We believe that evaluation of these changes would be helpful in diagnosing MDS.
The relationship between chronic lymphocytic leukaemia and prolymphocytic leukaemia
Authors: Melo Junia V. , Catovsky Daniel , Galton David A. G.
Published: March 2008

Summary. A series of 55 patients with B‐cell chronic lymphocytic leukaemia (CLL) and prolymphocytic leukaemia (PLL) was studied in order to investigate the evolution of prolymphocytoid transformation of CLL. Peripheral blood films were assessed for the percentages of prolymphocytes (%PROL) during a follow‐up period of 2 months to 24 years. The majority of patients with typical CLL (≤ 10% PROL) had minor variations or transient increases in %PROL throughout the course of the disease, but in one third of them a steady rise in the proportion of these cells was documented. Patients presenting with 11–55% PROL, referred to as CLL/PL, exhibited three patterns of evolution: half of them showed stability of the PROL counts, one third had unsustained increases in the %PROL and 18% showed definite progression to a PLL‐like blood picture. Patients diagnosed as FLL (> 55% PROL) had maintained a high %PROL during the period of observation. Serial marker studies in some CLL and CLL/PL cases showed that the percentage of M‐rosettes and the intensity of Smlg remained at the same initial levels in all but two cases. These two cases in the CLL/PL group showed a significant decrease in the percentage of M‐rosettes and stronger Smlg staining associated with progressive prolymphocytoid transformation. Patients with CLL and CLL/PL who had a sustained rise in the %PROL responded poorly to treatment compared with those with stable disease.
Different platelet specificities of heparin‐dependent platelet aggregating factors in heparin‐associated immune thrombocytopenia
Authors: Pfueller Sharron L. , David Rosemary
Published: March 2008

Summary. Delayed onset heparin‐associated thrombocytopenia (HAT) is thought to be a result of formation of antiplatelet antibodies which cause platelet aggregation in the presence of heparin. Platelet aggregation in response to serum from patients with HAT has been studied in platelet‐rich plasma (PRP) from a panel of normal blood donors. Heparin‐dependent aggregation with any HAT serum occurred in PRP from only some donors. PRP from the non‐responding donors did, however, aggregate in the presence of heparin with other HAT sera. The same patterns of aggregation or lack of response to HAT sera were seen in washed platelet suspensions. Heparin (0.06–2 U/ml) did not cause aggregation in the presence of normal serum with PRP from these donors. However, in PRP from four of the 17 individuals studied, heparin (0.25–1 U/ml) alone caused rapid platelet; aggregation and some HAT sera heated at 56°C caused platelet aggregation without added heparin. Sub‐aggregating concentrations of adrenaline could replace heparin in promoting aggregation by heated HAT sera in PRP of the other donors. HAT IgG showed the same platelet specificities as the serum in causing either heparin‐ or adrenaline‐dependent aggregation. Thus in HAT, antibodies are directed towards different platelet antigens which are expressed differently in different individuals. Platelet activation by heparin and adrenaline either exposes these antigens or causes aggregation of antibody‐coated platelets.
Expression of HPCA‐1 and HLA‐DR antigens on growth factor‐ and stroma‐dependent colony forming cells
Authors: Watt Suzanne M. , Katz Fay E. , Davis Laura , Capellaro David , Gordon Myrtle Y. , Tindle Robert W. , Greaves Melvyn F.
Published: March 2008

The expression of HLA‐DR and HPCA‐1 antigens (recognized by the L243 and BL3C5 antibodies respectively) on adult human bone marrow cells was examined by fluorescence activated cell sorting and colony assays. Nearly all the (day 14) lineage restricted and multipotential colony forming cells analysed in methylcellulose cultures in the presence of added growth factors express HLA‐DR and HPCA‐1 determinants. Two colour cell sorting reveals that the lineage restricted HLA‐DR positive progenitors express variable levels of BI.3C5 positivity whereas most of the multipotential progenitors, the multi‐CFC or CFU‐GEMM, are highly BI.3C5 positive. The isolated HLA‐DR and BI.3C5 positive populations also contain haemopoietic precursors which adhere to and form colonies on pre‐formed stromal layers. Thus, haemopoietic progenitors assayed in both types of culture system can be analysed and enriched by simultaneous two‐colour sorting using anti‐HLA‐DR and BI.3C5 monoclonal antibodies. Similarities in the antigenic phenotype of such cells, however, precludes the use of these reagents for segregating growth factor‐dependent from stroma‐dependent progenitors.
Improved in vivo and in vitro viability of platelet concentrates stored for seven days in a platelet additive solution
Authors: Holme S. , Heaton W. A. , Courtright M.
Published: March 2008

An additive solution has been developed for storage of platelet concentrates (PC) which sustains improved in vivo and in vitro viability after 7 d of storage in second generation oxygen permeable containers. This platelet additive solution is a protein‐free physiologic salt solution fortified with citrate, bicarbonate and glucose. The in vivo quality of the PC was evaluated by autologous radiolabelling with Indium‐111‐oxine to measure recovery and survival by multiple hit analysis. The in vitro quality was evaluated by total ATP content, hypotonic shock response and extent of shape change with ADP. Ten paired studies were performed with PC from the same donor being stored for 7 d at 22°C in both CPDA‐1 plasma and the additive solution. Mean recoveries and survivals were found to be substantially higher with PC stored in the additive solution than with PC stored in CPDA‐I plasma (51·0±7·8% and 144·1 ± 15·9 h versus 36·6±10·7% and 110·4±31·6 h). The differences were statistically significant (P< 0·001). The results of the in vitro assays described above parallelled the in vivo results, with statistically significantly superior results (P<0·01) for all parameters of PC stored in the additive solution. This study is the first to show that PC quality may be improved and storage extended using an additive solution.
Prothrombin Poissy: a new variant of human prothrombin
Authors: Dumont M.‐D. , Tapon‐Bretaudiere J. , Fischer A.‐M. , Bros A. , Chassevent J. , Aufeuvre J.‐P.
Published: March 2008

A new congenital dysprothrombinaemia is described in a newborn baby girl who presented severe bleeding from the second day of life. Routine coagulation tests showed very prolonged prothrombin time and activated partial thromboplastin time with about 2% prothrombin activity in a one‐stage assay. Staphylocoagulase and Echis carinatus venom prothrombin assays were respectively 35% and 25%. The prothrombin antigen level was 47% and its migration in crossed immuno‐electrophoresis was abnormal. Family study revealed the presence of both normal and abnormal prothrombin in the plasma of three family members: the father, the mother and the brother. Thrombin generation in a system free from natural inhibitors showed that the abnormal prothrombin was slowly and incompletely activated. The propositus is thought to be homozygous for a ‘lazy’ dysprothrombin.
Cianidanol and its metabolites bind tightly to red cells and are responsible for the production of auto‐ and/or drug‐dependent antibodies against these cells
Authors: Salama A. , Mueller‐Eckhardt C.
Published: March 2008

Cianidanol ((+)‐2‐(3,4‐dihydroxyphenyl)‐3, 5, 7‐chromantriol) is a flavonoid which has been associated with severe immune haemolysis by as yet unclear mechanisms. We report six patients who developed haemolysis while receiving the drug. The disorder was episodic in all patients and resolved after discontinuing the drug. The causative antibodies could be demonstrated in all six cases, even when the haemolytic episode was more than 1 year prior to this study. One patient had developed drug‐independent IgG autoantibodies, another simultaneously developed autoantibodies and drug‐dependent antibodies (ddab) of the IgG class, while the remaining four patients had only ddab of the IgM and/or the IgG classes. All ddab were reactive with red blood cells (RBC) in the presence of the drug and/or its metabolites (ex vivo antigens), and, quite unexpectedly, also with RBC coated with the drug (metabolites) in vitro or in vivo. This reactivity did not change either by preincubating the antibodies with the drug or by adding large amounts of the drug to the mixture of drug‐coated cells plus antibody. It seems that the stable association of cianidanol with RBC generates antigenic sites against which a heterogeneous immune response is elicited giving rise to long‐lasting drugdependent antibodies as well as autoantibodies.
Clinical and laboratory features of de novo acute myeloid leukaemia with trilineage myelodysplasia
Authors: Brito‐Babapulle F. , Catovsky D. , Galton D. A. G.
Published: March 2008

Primary myelodysplastic syndromes progress to acute myeloid leukaemia (AML) in about 30% of cases. We have sought evidence of pre‐existing trilineage myelodysplasia (TMDS) using the FAB criteria (1982) in 160 consecutive cases of primary de novo AML. TMDS was found in 24 cases (15%) including two of 33 cases of Ml (6%), four of 40 cases of M2 (10%), none of 18 cases of M3, five of 31 cases of M4 (15%), six of 30 cases of M5 (20%), all of six cases of M6 and one of two cases of M7.

The median presentation bone‐marrow blast‐cell count in the 24 AML/TMDS cases was 53% (30–90%) and 82% (45–100%) in the 136 cases of AML without TMDS. 60% of the AML/TMDS bone‐marrow aspirates contained fewer than 60% of blasts compared with only 11% of those from AML without TMDS (P<0.001). In AML the occurrence of symptomatic cytopenias when the marrow blast‐cell count is below 60% and the peripheral blood blast‐cell count is below 20% is highly correlated with dysplastic haemopoiesis (P<0.001).
Chromosome abnormalities in peripheral T‐cell lymphoma
Authors: Lakkala‐Paranko Taina , Franssila Kaarle , Lappalainen Kimmo , Leskinen Ritva , Knuutila Sakari , de la Chapelle Albert , Bloomfield Clara D.
Published: March 2008

We have studied neoplastic lymph nodes from six patients histologically, immunologically and cytogenetically. Histologically all the cases were classified as peripheral T‐cell lymphomas. These were subclassified as T‐zone lymphomas in three, large cell ‘pale cell’ variant in one. large cell immunoblastic in one, and small cell, mycosis fungoides in one. Two had features of angioimmunoblastic lymphadenopathy (AILD). Immunologically all cases expressed CD2 (OKT11) and CD4 (T4). All six cases had clonal chromosome abnormalities, although in four cases the majority of cells were chromosomally normal. Chromosome 3 was most often involved in abnormalities, occurring in five patients. The most common single chromosome abnormality, trisomy 3, was seen in all three cases classified as T‐zone lymphoma and in no other cases. In the two cases with features of AILD only numerical abnormalities were seen, whereas in the other cases complicated structural rearrangements were present. Recurring structural abnormalities involved bands 1p12or13, 1q32, 3p25 and 14q11. Our data suggest that cytogenetic analysis may assist in diagnosis and classification of the peripheral T‐cell lymphomas.
Identification of blast cells in peripheral blood through automatic assessment of nuclear density: a new tool for monitoring patients with acute leukaemia
Authors: D'Onofrio G. , Mancini S. , Leone G. , Bizzi B. , Mango G.
Published: March 2008

The Technicon H*1 haematology system is provided with a new method for basophil count, neutrophil lobularity assessment and detection of blasts in peripheral blood through automated measurement of nuclear density. We compared the results of the H* 1 blast flag with those of the microscope examination in 131 peripheral blood samples from 43 patients with acute leukaemia in different phases of their disease, to determine the degree of sensitivity and specificity of the system in this setting. In six patients at diagnosis or in overt relapse, all having large percentages of blasts at the manual differential count, a typical deformation of the profile of the mononuclear cell population on the display was consistently observed, regardless of the morphological subtype of leukaemic cell which was involved. Amongst 34 samples with 4–95% morphologically recognizable blasts on the peripheral blood film, the sensitivity of the system was 100%. with no false negatives at all. In 43 samples, on the other hand, the H*1 blast flag was positive in the absence of any morphological evidence of blasts on the smear. These ‘false positives’, however, were always obtained from leucopenic patients who had more than 12% blast infiltration in the bone marrow, compared to the 0–6% value that was found in patients with a negative H*1 blast flag. These results suggest that the H*1 system is a highly sensitive tool for the detection in peripheral blood of even small concentration of leukaemic cells, which escape morphological identification.
Haemolytic ‘efficiency’ of C5b‐9 complexes in drug‐induced immune haemolysis: role of cellular C5b‐9 distribution
Authors: Salama A. , Mueller‐Eckhardt C. , Boschek B. , Bhakdi S.
Published: March 2008

Summary. We report on quantitative analyses of C5b‐9 binding to target red blood cells (RBC) lysed through the action of drug (nomifensine)‐dependent antibodies (ddab) and anti‐RBC antibodies (warm plus cold agglutinins). Immunoradiometric assays showed that, at any given degree of haemolysis, more C5b‐9 was bound to cells sensitized with anti‐RBC antibodies compared to cells lysed via ddab. Thus, C5b‐9 complexes deposited through the action of ddab appeared to be haemolytically more ‘efficient’ than those deposited via anti‐RBC antibodies. However, data obtained from flow‐cytometric assays and immunoelectron micros‐copy demonstrated a major role for the distribution (total number) of C5b‐9 lesions among the cells within a given cell population. Enhanced haemolysis with a given total amount of C5b‐9 complexes appeared to be derived from a more even distribution of these complexes among the cells rather than from higher lytic efficiency of individual complexes. Since ddab bind with low affinity to cells, we suggest that these antibodies diffuse from cell to cell and cause extensive haemolysis through the deposition of relatively few C5b‐9 complexes at each site. In contrast, high affinity antibodies remain largely bound to relatively restricted sites causing deposition of larger amounts of C5b‐9 on, and hence lysis of, numerically fewer cells. With respect to the net intravascular haemolytic effect, our findings may explain why comple‐ment‐activating antibodies of low affinity are often more detrimental than high affinity antibodies.
Detection of the Tn antigen in leukaemia using monoclonal anti‐Tn antibody and immunohistochemistry
Authors: Roxby David J. , Morley Alexander A. , Burpee Marie
Published: March 2008

Exposure of the normally cryptic Tn antigen on haemopoietic cells leading to erythrocyte polyagglutination has been reported in a few cases of malignant or premalignant haemopoietic disorders and has been attributed to a selective deficiency of the enzyme 3–β‐D‐galactosyl‐transferase, A male patient presented with acute myelomonocytic leukaemia with no evidence of Tn expression but, 16 months later, in the terminal stage of the disease, the majority of the erythrocytes were found to be polyagglutinable. Tn expression was confirmed by the use of lectins and by agglutination with a Tn‐specific monoclonal antibody. FBT3. Retrospective studies of stored blood and bone marrow smears were performed by immunocytochemistry using FBT3. Tn positive cells were first detected in the marrow 8 months prior to death. They increased progressively in number and, in the terminal illness, over 90% of erythroid precursors in the marrow and erythrocytes in peripheral blood and 40% of granulocyte precursors of the marrow and 10% of granulocytes in the blood were Tn positive. These observations suggest that Tn expression was present in a subclone of cells which became dominant during the course of the disease and that there may be a relationship between Tn expression and leukaemic progression.
Interpretation of red cell survival data for in vivo compatibility testing: a normal value study
Authors: Bentley S. A. , Collins M. L. , Miller D. T.
Published: March 2008

Short‐term survival studies are sometimes required to determine the compatibility of donor red cells. The results of these studies are generally expressed as per cent survival at 60 min. The present study was undertaken to investigate the potential for more sophisticated data analytical techniques to improve sensitivity.

In one group of eight healthy male volunteers, autologous red cells were labelled with 51chromium and injected immediately, while in a second group, red cells were stored for 5 d prior to injection. In both groups, eight to 10 samples were collected in the first 6 h and another 10–12 samples over the next 4 weeks. Estimation of the 60 min per cent survival was insufficiently sensitive to detect ‘physiological’haemolysis following injection of 5‐d‐old autologous blood. Regression analysis of 6 h survival data, however, demonstrated significantly higher red cell clearance rates in these cases than in those receiving fresh cells, with a mean 24 h loss of 3.3% of activity. The upper limit for the 6 h red cell clearance rate was 1.63%/h after fresh autologous blood and 2.43%/h after 5‐d‐old blood. The significance of these findings is discussed and a protocol suggested for the analysis of short‐term red cell survival data.
Myeloid progenitors from the bone marrow of patients with vitamin D resistant rickets (type II) fail to respond to 1,25(OH)2D3
Authors: Nagler Arnon , Merchav Shoshana , Fabian Ina , Tatarsky Ilana , Weisman Yosef , Hochberg Zeev
Published: March 2008

The active metabolite of vitamin D, 1,25‐dihyd‐roxyvitamin D3 (1,25(OH)2D3), has been shown to enhance the growth of human granulocyte/macrophage haemopoietic progenitors in vitro and to induce these cells to differentiate along the monocyte/macrophage pathway. In order to evaluate the relationship between specific receptors for 1,25(OH)2D3 and the role of 1,25(OH)2D3 in the regulation of haemopoietic cell differentiation, we examined the effect of 1,25(OH)2Di on the in vitro growth and differentiation patterns of marrow myeloid progenitor cells from two patients with 1,25(OH)2D3 resistant rickets, resulting from defective receptors to vitamin D. A significant rise in the frequency of myeloid colonies in control marrow cell cultures was induced by 2×10−9 to 2×10−7 M 1,25(OH)2D3. This rise reached a plateau at 2×10−9–2×10−8 M 1,25(OH)2D3, resulting in a maximal 54±9% increase in colony numbers. In contrast, no stimulatory effect could be detected when 1,25(OH)2D3 was added to cultured marrow cells from the patients with 1,25(OH)2D5 resistance. Analysis of colony composition revealed that 2×10−8 and 2×10−7 M, 1,25(OH)2D3 induced a 50±26% increase in the frequency of colonies composed only of monocytes/macrophages in control, but not in the patients’marrow cell cultures. The effect of 2×10−8 and 2×10−7 M 1,25(OH)2D3 on progenitor cell differentiation towards monocytes/macrophages was also observed in marrow cell suspension cultures. Whereas 1,25(OH)2D3 induced a 81–136% increase in the frequency of monocytes in control marrow cells, no effect could be detected on the generation of mature monocytes in marrow cells of the 1,25(OH)2D3 resistant patients. Our results show that marrow granulocyte/macrophage progenitor cells from patients with 1,25(OH)2D3 resistance fail to respond to 1,25(OH)2D3. We thus demonstrate that the effect of 1,25(OH)2D3 on the proliferation and differentiation of haemopoietic progenitor cells is mediated through its binding to specific cytoplasmic receptors.
Interleukin 2 enhances cytotoxic cell function in vitro after T‐cell depleted marrow transplantation
Authors: Leger Olivier , Drexler Hans G. , Reittie Joyce E. , Secker‐Walker Lorna , Grant Prentice H. , Brenner Malcolm K.
Published: March 2008

After T‐cell depleted marrow transplantation, there is a rapid recovery of cytotoxic effector cells, with activity against targets not susceptible to killing by ‘resting’natural killer cells. These targets include Epstein‐Barr virus transformed B cells and leukaemic cell lines. Activated killer cell function declines by 3 months after transplantation. We find that when CD3 negative effector cells are obtained from these patients and cultured in vitro with interleukin 2 there is a further enhancement of cytotoxic activity against a range of target cells in the early post‐transplant period. and a restoration of high level cytotoxic activity to effector cells obtained 3 months or more after the procedure. These results may have relevance to attempts to reduce the incidence of leukaemic relapse, and EBV + ve lymphoma outgrowth after T‐cell depleted BMT.
Multiple molecular abnormalities in Ph1 chromosome positive acute lymphoblastic leukaemia
Authors: Dreazen Orna , Klisak Ivana , Jones Gary , Ho Winston G. , Sparkes Robert S. , Gale Robert Peter
Published: March 2008

The Ph1 chromosome is present in 95% of patients with chronic myelogenous leukaemia (CML). The Ph1 chromosome also occurs in 5–25% of children and adults with acute lymphoblastic leukaemia (ALL). This observation raises questions as to whether these diseases are similar or identical. In patients with CML the c‐abl and hcr genes are translocated and abnormally expressed. We studied molecular events related to bcr and c‐abl in five patients with ALL to determine its relationship to CML. Four had the Ph1 chromosome; the fifth a probable Ph1 chromosome. c‐abl and bcr abnormalities identical to CML were detected in four suggesting a common molecular basis. One patient with the Ph1 chromosome and c‐abl translocation lacked these molecular changes but had abnormal c‐abl gene transcription apparently unrelated to her. These data suggest that Ph1 chromosome positive ALL is heterogeneous; in some patients the molecular abnormality is identical to CML; in others c‐abl is likewise involved but via a different mechanism.
Neutrophil FC‐receptor expression in cells of the circulating and marginated pools
Authors: Baglin Trevor P. , Pollock Andrew
Published: March 2008

Summary. A method for rosetting neutrophils was developed and optimal rosetting conditions determined. The adhesive properties of neutrophils and their ability to form rosettes were studied for cells in the circulating pool, and following exercise when the circulating pool had been enriched by neutrophils from the marginated pool. We found the marginated pool to be the same as the circulating pool in terms of rosetting subpopulations and the in vitro adhesive properties of these subpopulations, indicating that the two pools can be considered kinetically as one.

We also showed that non‐rosetting neutrophils can develop the ability to form rosettes following adhesion to glass in the presence of autologous serum.
Erythropoiesis during an erythroblastic phase of chronic myeloproliferative disorder associated with monosomy 7
Authors: Kojima Seiji , Mimaya Junichi , Tonouchi Tsutomu , Yokochi Tsunehiro , Kajitani Sachiko
Published: March 2008

Summary. A chronic myeloproliferative disorder associated with monosomy 7 is described in a 3 1/2‐year‐old boy. His presenting features closely resembled those of juvenile chronic myeloid leukaemia (JCML). Cytogenetic study of bone marrow cells showed that all of the metaphases examined had chromosome 7 deletions.

He developed an erythroblastic phase, characterized by anaemia, marked erythroid hyperplasia of bone marrow and the appearance of nucleated red blood cells in the peripheral blood. During the erythroblastic phase, blood transfusion resulted in a suppression of erythropoiesis as evidenced in both the peripheral blood and bone marrow. The in vitro culture studies showed that the erythroid precursor was dependent upon erythropoietin (Ep) for differentiation and proliferation during the erythroblastic phase. However, the Ep dose‐response curve showed that a peak of erythroid colony formation occurred at a lower concentration than in the healthy controls. These findings suggest that although the erythroid precursor remains under the control of Ep, it has an increased sensitivity to Ep during the erythroblastic phase of monosomy 7.
Hybrid leukaemia of T cell and myeloid lineages: cytogenetic distinction from second (induced) malignancy
Authors: Simpson E. M. , Mott M. G.
Published: March 2008

Summary. The therapeutic and prognostic implications of relapse and clonal evolution of leukaemia are substantially different from those of secondary (induced) malignancy. This report documents the case of a patient who presented with apparent acute non‐lymphocytic leukaemia (ANLL) following therapy for acute (T‐cell) lymphoblastic leukaemia (ALL) 4 years previously. Morphologically and cytochemically the cells were of myeloid type, but the cell markers showed a T cell lineage. Cytogenetic studies confirmed that this was a relapse of T cell ALL with a phenotypic change, rather than a second malignancy induced by chemotherapy. A 14q deletion present at initial diagnosis recurred at relapse, with the addition of cells with complete deletion of chromosome 14, indicating clonal evolution.
Abnormal proliferation of T‐colony‐forming cells from peripheral blood of patients with T‐cell acute lymphoblastic leukaemias and lymphomas in complete remission: potential prognostic value
Authors: Allouche Michéle , Georgoulias Vassilis , Bourinbaiar Aldar , Dupuy Isabelle , Clemenceau Corine , Gaget Hélène , Mathe Georges , Jasmin Claude
Published: March 2008

Summary. Peripheral blood T‐colony‐forming cells (T‐CFC) from most patients with T‐cell acute lymphoblastic leukaemias (T‐ALL) and T‐cell non‐Hodgkin's lymphomas (T‐NHL), can proliferate in vitro in methylcellulose in the absence of added growth factors or mitogens. We now report that spontaneous T‐cell colonies could also be obtained during complete remission of 13 out of 21 patients with T‐ALL and T‐NHL, but none of eight patients with common (pre‐B) ALL (cALL). Colony cells were mainly E+T3+, with a variable expression of other T cell markers. Spontaneous T‐CFC did not possess self‐renewal capacity in the absence of added growth factors. Moreover, incubation of spontaneous colonies with colchicine yielded mitoses in only two out of seven patients, with one normal and one abnormal karyotype. In five patients tested, recombinant interleukin 2 (IL2) could also induce the proliferation of some T‐CFC. Both spontaneous and IL2‐induced colonies were inhibited by an anti‐IL2 receptor monoclonal antibody, suggesting that interaction of IL2 with its receptor may be involved in the proliferation of some T‐CFC from these patients.

A study of 14 T‐ALL patients tested during their first remission indicated that patients who developed no or few spontaneous colonies during their first remission (less than 20 colonies/105 mononuclear cells) seemed to relapse later and to have a significantly longer survival than patients with a high number of spontaneous colonies. These data suggest that the spontaneous proliferation capacity of T‐CFC might be of prognositic value in the clinical evaluation of T‐ALL.
Grey platelet syndrome: studies on platelet alpha‐granules, lysosomes and defective response to thrombin
Authors: Srivastava P. C. , Powling M. J. , Nokes T. J. C. , Patrick A. D. , Dawes Joan , Hardisty R. M.
Published: March 2008

Summary. The platelets of a young man with the grey platelet syndrome were severely depleted of all seven α‐granule proteins assayed as well as partially deficient in α‐mannosidase and α‐fucosidase; four other lysosomal enzymes were present in normal concentrations. Total platelet 5‐hydroxy‐tryptamine (5HT) and adenine nucleotides were normal, and14C‐5HT uptake reached normal levels only slightly more slowly than a control. Aggregation and dense body secretion occurred normally in response to ADP, adrenaline, collagen, PAF‐acether, sodium arachidonate, A23187, lonomycin, TPA and U44069, but were very delayed in response to thrombin. The increase in cytosolic free calcium in response to thrombin was very slow and much reduced in amplitude, whether in the presence or absence of extracellular Ca2+. These defects in response to thrombin were not corrected by the separate addition of purified α‐granule proteins or by a whole releasate from normal platelets. It is suggested that these platelets, in addition to their α‐granule deficiency, may have a specific defect of thrombin receptor‐mediated activation of phospholipase C.
Detection of circulating human platelet fragments by using monoclonal antibodies against platelet glycoprotein IIb‐IIIa complex
Authors: Vos J. J. E. , Huisman J. G. , Modderman P. W. , Kr. A. E. G. , Borne Von Dem
Published: March 2008

Summary. A radioimmunoassay (RIA) for the detection of platelets and platelet fragments was developed. A sandwich of two monoclonal antibodies directed against the platelet‐specific glycoprotein complex IIb‐IIIa (GP IIb‐IIIa) was used in this assay. A discontinuous 7·5‐20% (v/v) albumin gradient was applied to separate platelets and their fragments of various sizes. In platelet suspensions fractionated in this way, we observed that particles smaller than normal platelets still carried the GP IIb‐IIIa antigens. This procedure enabled us to detect platelet‐derived particles in platelet‐rich plasma from thrombocytopenic patients.
Anticardiolipin antibodies associated with HTLV‐III infection
Authors: Canoso Rosa T. , Zon Leonard I. , Groopman Jerome E.
Published: March 2008

Summary. Anticardiolipin antibody levels were determined in 73 homosexual men. Thirty of these patients had acquired immunodeficiency syndrome (AIDS), 16 patients had the AIDS‐related complex (ARC) and 27 were healthy. Antibodies to human T‐lymphotropic virus type III (HTLV‐III) were detected in all patients with AIDS and ARC and in 11 of the healthy homosexuals. Eight patients with positive fluorescent treponemal antibody absorption test were excluded from the study. High levels of IgG‐anticardiolipin antibodies were present in 23 of the 28 patients with AIDS; 12 of the 14 with ARC; five of the 10 HTLV‐III positive healthy homosexuals; and none of the 13 HTLV‐III negative healthy homosexuals. High levels of IgM‐ACA were detected only in four patients with AIDS. The IgG‐anticardiolipin levels were higher in the HTLV‐III positive patients than in the HTLV negative group (P=0·013). None of the patients with anticardiolipin antibodies exhibited venous thrombosis.