bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of Chloroquine‐Treated Granulocytes and Platelets in the Diagnosis of Immune Cytopenias
Authors: Metcalfe P. , Minchinton R.M. , Murphy M.F. , Waters Prof.A.H.
Published: March 2009

A chloroquine modification of the fluorescent antiglobulin technique has been used to demonstrate cell‐specific antibodies in the presence of HLA antibodies. This is of particular value in the diagnosis of alloimmune neonatal thrombocytopenia and neutropenia, post‐transfusion purpura, and in the investigation of febrile non‐haemolytic transfusion reactions and of patients refractory to platelet transfusions.
Macroaggregate Formation in Optimal Additive Red Cells
Authors: Robertson M. , Boulton F. E. , Doughty R. , MacLennan J. R. , Collins A. , McClelland D. B. L. , Prowse C. V.
Published: March 2009

The formation of macroaggregates weighing up to 9 g was observed in optimal additive red cells. Such aggregates, with a mean wet weight of 3 g, formed progressively during storage and were present in up to 85% of units. They were composed of leucocyte and platelet debris, together with some fibrin. Macroaggregate formation was halved by less stringent centrifugation during preparation and was reduced by use of an optimal additive system in which the additive solution contained citrate. Extra mixing during transfer of the additive solution only delayed aggregate formation. Partial leucocyte depletion or addition of 200,000 KIU of the enzyme inhibitor aprotinin did not prevent macroaggregate formation.
A New Red Blood Cell Cold Autoantibody (Anti‐Me)
Authors: Salama A. , Pralle H. , Mueller‐Eckhardt C.
Published: March 2009

A new red blood cell (RBC) cold autoantibody (anti‐Me) is presented which was detected in a patient with autoimmune hemolytic anemia of ‘cold type’ associated with Waldenström's macroglobulinemia. The autoantibody was of the IgM (Kappa) class, had a moderate titer, and reacted with cord as well as with adult RBC. Its hemagglutinating activity against RBC was markedly increased in the presence of preheated human milk, but not in the presence of simple sugars or preheated casein. The data suggest that the antibody is directed against a new (glycolipid) antigen common to human RBC termed Me antigen (‘milk enhanced’).
Current Issues in Neonatal Transfusions
Authors: Strauss Ronald G.
Published: March 2009

Abstract. Two controversial issues of neonatal transfusion practices, erythrocyte ‘booster’ transfusions and granulocyte transfusions, are critically reviewed, and current recommendations for transfusion practices are made. Infants should receive erythrocyte transfusions to treat congestive heart failure caused primarily by anemia. It is customary to maintain the hematocrit at > 40% in neonates with severe respiratory disease, although the efficacy of this practice has not been firmly established. Erythrocyte transfusions seem to be indicated for infants with anemia plus recurrent apnea, poor weight gain or the syndrome of tachycardia, tachypnea, dyspnea and poor feeding for which no other cause can be found. Granulocyte transfusions are likely to benefit seriously ill neonates exhibiting all three of the following: strong evidence of bacterial sepsis, neutropenia (compared to age‐related normal values) and a diminished marrow neutrophil storage pool. Granulocyte transfusions for septic infants expressing only one or two of these features should be considered to be experimental therapy.
An Lu(a‐b‐) Phenotype Caused by an X‐Linked Recessive Gene
Authors: Norman P.C. , Tippett P. , Beal R. W.
Published: March 2009

Abstract. The family of a male Lu(a‐b‐) propositus whose red cells showed some characteristics of the dominant and some of the recessive type of Lu(a‐b‐) phenotype was studied. The investigation revealed a third genetic background for this rare phenotype, an X‐borne suppressor recessive in effect. The notation proposed is XS1 for the normal gene and XS2 for the suppressor gene which affects the expression of Lutheran and other antigens.
An Infant Girl with Severe Autoimmune Hemolytic Anemia: Apparent Anti‐Vel Specificity1
Authors: Becton David L. , Kinney Thomas R.
Published: March 2009

Abstract. Autoantibodies directed against the common red‐cell antigen Vel are unusual, with only 2 previous cases reported. This report describes an infant girl with steroid‐resistant autoimmune hemolytic anemia whose serum contained an antibody with apparent specificity for Vel antigen. Transfusion with Vel‐positive cells resulted in a massive hemolytic transfusion reaction. Survival of labelled Vel‐negative cells was normal up to 8 h following transfusion, and no transfusion reaction followed subsequent use of Vel‐negative cells. There was no evidence of a collagen vascular disease, an immunodeficiency syndrome, or a malignancy. Although rare, anti‐Vel should be considered in unexplained autoimmune hemolytic anemia.
RASM, a ‘New’ Low‐Frequency Blood Group Antigen
Authors: Brown A. , Plantos M. , Moore B.P.L. , Jones T.
Published: March 2009

Abstract. A ‘new’ low‐frequency red cell antigen, RASM, was found in 3 generations of a Caucasian family. It is inherited as a Mendelian dominant character. The propositus, a newborn, had a positive direct antiglobulin test; overt haemolytic disease of the newborn was not present.
Filterability of ACD‐Stored Red Cells
Authors: Galea George
Published: March 2009

Abstract. The deformability of stored erythrocytes was studied using the Myrenne filtrometer MF4. Blood was stored in ACD under usual blood banking conditions and filterability studies were done at various intervals during a 3‐week period. Marked reductions in stored erythrocyte filterability were noted as early as day 5 of storage. These results show that ACD is not a good ‘rheological’ storage medium and indicate that further studies are required, using different storage media to assess the importance and significance of this rheological lesion.
Continuous Monitoring of Platelet Morphology during Small‐Scale in vitro Storage
Authors: Trenchard P.M.
Published: March 2009

Abstract. A commercially available aggregometer was mechanically and electronically modified to enable automatic repeated measurements of platelet morphology for platelet suspensions undergoing prolonged storage within a sealed perspex cuvette, at any temperature. The modifications are described in sufficient detail to enable others to readily produce the same or similar apparatus. Representative experiments with such apparatus are described, and indicate its potential usefulness in defining the rate of change of platelet morphology occurring during storage at any given temperature, e.g. progressive alteration of shear‐induced light transmission change values.
A Human‐Human Monoclonal Anti‐D by Direct Fusion with a Lymphoblastoid Line1
Authors: Lowe A.D. , Green S.M. , Voak D. , Gibson T. , Lennox E.S.
Published: March 2009

Abstract. The human lymphoblastoid cell line W1‐L2–729‐HF2 has been fused with B cells from a plasmaphoresed anti‐D donor immunized with D+ cells. A stable monoclonal antibody‐producing cell line has been produced which yields culture supernatant of good titre without the need for concentration. The production and use of this reagent as an alternative Rh D typing reagent for use by saline and enzyme manual tests and automated tests in a Technicon 16C machine is discussed. Du red cells are detected in enzyme enhanced tests.
Thermal Inactivation of Human Immunodeficiency Virus in Lyophilised Blood Products Evaluated by ID50 Titrations
Authors: Tersmette Matthys , de Goede Ruud E. Y. , Over Jan , de Jonge Egge , Radema Horst , Lucas Cornells J. , Huisman Han G. , Miedema Frank
Published: March 2009

Abstract. Inactivation of human immunodeficiency virus (HIV) in lyophilised small pool cryoprecipitate, factor VIII concentrate, prothrombin complex and C1 ‐esterase inhibitor concentrate by prolonged heat treatment (72 h, 60° C) was studied. Plasma products, inoculated prior to lyophilisation, had infectious titres ranging from 107 to 1010.5. Residual infectivity (TCID50) was assessed by multiple titrations on H9 cells in a macro system and subsequent detection of virus replication by determining reverse transcriptase activity. Kinetics of inactivation showed a biphasic pattern: during the first 8 h a variable TCID50 reduction up to 104.3 was observed, followed by an additional loss of 101–102.7 during the next 64 h. Heat treatment for 72 h resulted in a mean TCID50 reduction of 105. It is concluded that prolonged heat treatment may lead to the adequate prevention of HIV transmission by lyophilised plasma products.
Measurement of Platelet Surface‐Bound IgG by a Monoclonal125I‐Anti‐IgG Assay
Authors: Court Wayne S. , LoBuglio Albert F.
Published: March 2009

Abstract.We have described the use of a monoclonal125I‐labeled anti‐IgG (125I‐MA) to assay IgG antibody displayed on the surface of platelets from normal and immune thrombocytopenic patients and reported levels of IgG 10–100‐fold lower than previous studies. This report describes the immunologic characteristics of the125I‐MA and the assay for surface IgG. The125I‐MA has a high binding affinity for surface‐displayed IgG (2.22 times 109M‐1), reacts equally well with all four subclasses of IgG and not at all with IgM or IgA. In our assay, the binding ofI25I‐MA was found to be ≥99% specific for IgG (no nonspecific association ofI25I‐MA with platelets) and the binding ratio ofI25I‐MA to IgG displayed on the cell surface was 0.91 (close to unity). Finally, platelet lysates were found to contain large amounts of IgG protein (39,597 ± 27,418 molecules/platelet) as compared to surface‐displayed IgG (124 ± 86 molecules/platelet). This assay has excellent characteristics for quantitation of IgG on platelets and the discrepancy with other techniques may, in part, be due to intentional or inadvertent lysis of platelets during assay conditions.
Occurrence and Characterization of Lympho‐Agglutinins in Indian Plants
Authors: Arora Jatinder S. , Sandhu Rajindar S. , Kamboj Sukhdev S. , Chopra Sushil K.
Published: March 2009

Abstract. Lympho‐agglutinins have been detected and characterized in 31 plant species. Out of these, 14 agglutinated only the neuraminidase‐treated cells. The lectin‐rich genera included Crotalaria and Erythrina (Fabaceae), Amaranthus (Amaranthaceae), Artocarpus (Moraceae) and Clerodendron (Verbenaceae). The new lectins varied in their potency and biological action spectra. The 3 Artocarpus species were found to be exceptionally potent and specific for melibiose, an α‐D‐galactoside. Among the most effective sugar inhibitors for other lectins were N‐acetyl‐galactosamine, lactose, galactose and asialofetuin/fetuin.
An Animal Model for the Detection of Hypotensive Side Effects of Immunoglobulin Preparations
Authors: Bleeker W. K. , Agterberg J. , Rigter G. , Rossen A. de Vries‐van , Bakker J. C.
Published: March 2009

Abstract. Intravenous administration of certain immunoglobulin preparations may cause severe adverse reactions, especially in immunodeficient patients. These reactions are generally assumed to be related to the anticomplementary activity of the preparations, caused by IgG aggregates. Because the exact mechanism of the adverse reaction is unknown, we investigated the reactions induced in anesthetized rats on rapid intravenous administration of different human immunoglobulin preparations. The most conspicuous observation was a severe, long‐lasting hypotension, induced by standard immunoglobulin preparations (for intramuscular use), which appeared to be independent from the concomitant complement and neutrophil activation. The long‐lasting hypotension was not related to the presence of prekallikrein activator, which induced a transient hypotensive reaction only after sensitizing the rats to bradykinin. The reactions appeared to be associated with IgG aggregates. It was found that certain aggregates induced mainly complement activation, whereas others had mainly a hypotensive effect or no effect at all. It was concluded that the rat model provides a sensitive and reproducible test system for vasoactive properties of immunoglobulin preparations for intravenous administration that cannot be predicted from in vitro measurements, such as anticomplementary activity, aggregate content or prekallikrein activator activity. It is suggested that the test may also be used for other plasma products for intravenous administration.
Human Monoclonal Antibodies to Rho(D)
Authors: Foung Steven K.H. , Blunt Judith A. , Wu Paul S. , Ahearn Patricia , Winn Linda C. , Engleman Edgar G. , Grumet F. Carl
Published: March 2009

Abstract. B lymphocytes from Rh negative donors with serum anti‐D antibodies were isolated and fused with the mouse‐human heteromyeloma, SBC‐H20, to produce hybridomas secreting IgM or IgG1 human monoclonal antibodies to D antigen. The IgM antibody in hybridoma supernatant agglutinates all normal D positive cells at the immediate spin phase of reactivity. Using concentrated IgM hybridoma supernatant of approximately 50 μg/ml, Du cells were also agglutinated. The IgG1 antibody reacts by indirect hemagglutination with all D and Du cells. Against Rh mosaics, different reactivity was noted for each antibody. Furthermore, D positive cells precoated with the IgG1 antibody inhibit the IgM direct hemagglutination, suggesting that the antibodies identify closely associated epitopes. These human monoclonal antibodies will be useful diagnostic reagents and, ultimately, should be useful in the prevention of Rh hemolytic disease of the newborn.
Competitive Enzyme‐Linked Immunoassay for the Quantitation of Platelet‐Associated Immunoglobulins (IgG, IgM, IgA) and Complement (C3c, C3d) with Polyclonal and Monoclonal Reagents1
Authors: Kiefel V. , Jäger Sigrun , Mueller‐Eckhardt C.
Published: March 2009

Abstract. A competitive enzyme‐linked immunoassay (CELIA) was developed for the quantitation of platelet‐associated immunoglobulins and complement proteins. The use of unlabeled polyclonal rabbit or monoclonal antibodies to human immunoglobulins and enzyme‐labeled anti‐mouse or anti‐rabbit IgG (double‐step technique) resulted in a higher sensitivity compared to the single‐step technique using only enzyme‐labeled anti‐human immunoglobulin antibody preparations. Sensitivity and results obtained by both techniques were compared. The range of normal values for platelet‐associated IgG, IgM, IgA, C3c and C3d was assessed upon a large number of normal blood donors. When platelet‐associated IgG was concomitantly assayed with polyclonal and monoclonal anti‐IgG by the double‐step technique on platelets obtained from normal donors and thrombocytopenic patients, identical results were obtained with both reagents. Problems related to the quantitation of immunoglobulins on platelets with different assays and antibody preparations are discussed.
Radio‐Immunoassay of the Functional Activity of Anti‐D (Rh) Preparations using a Human Monoclonal 125I‐Labelled Anti‐D
Authors: Hughes‐Jones N.C. , Dawes B. , Gorick B.D. , Melamed M.D. , Pepper R. , Thompson K.M.
Published: March 2009

Abstract. The prophylactic effect of anti‐D in the prevention of maternal immunization to the D antigen of the Rh system depends in the first instance on its ability to bind to fetal Rh‐positive red cells. The functional activity of the injected anti‐D in this respect is dependent on both the plasma concentration and functional affinity constant of the antibody, and both factors should be taken into account in assessing IgG anti‐D preparations for clinical use. A radio‐immunoassay utilizing 125‐I‐labelled human monoclonal anti‐D has been developed which measures the amount of antibody bound to red cells after the addition of an aliquot of the test anti‐D preparation. The assay is thus a functional test in that it measures the ability of the anti‐D to react with the antigen. The functional activity of the test anti‐D is compared to that of the international standard, and the concentration is then expressed in terms of μg‐equivalents, that is, its equivalence in functional terms to the stated amount of international standard. The radio‐immunoassay was compared to the currently used agglutination assay, and the results were found to give good agreement in 18 out of the 21 samples assayed.
Preparation of Leukocyte‐Poor Platelet Concentrates from Buffy Coats
Authors: Pietersz R.N.I. , Reesink H. W. , Dekker W.J.A.
Published: March 2009

Abstract. Six citrate phosphate dextrose (CPD)‐saline adenine glucose mannitol (SAG M) quadruple systems were evaluated for the preparation and storage of leukocyte‐poor platelet concentrates (PC) from buffy coats. The platelet storage bags examined were manufactured from normal polyvinylchloride (PVC) or special‐type plastics. Biotest supplied PVC 76 (n= 14) and PVC 763 (n=16) NPBI supplied PSV 3277 (n= 15) and DPL‐110 (n= 14) and Terumo supplied Teruflex (n=18) and molded Teruflex (n=14). The PC were stored for 7 days at 22 °C on a horizontally shaking platform. Cell counts, pH, PO2, PCO2, morphology score and swirling patterns were monitored at 5, 72, 120 and 168 h. The plasma volumes averaged 63 ml and ranged from 39 to 81 ml, the overall mean ±SD platelet concentration was 0.89±0.33 × 109/ml. None of the PC had a leukoyte count higher than 10× 106 per unit. After storage for 168 h, the pH ranged from 6.56 to 7.40 for all brands. The PO2 remained stable and even rose significantly (p<0.05) during storage in the NPBI PSV 3277 and Terumo molded Teruflex bags. The PCO2 decreased equally in all bags. Morphology scores were well maintained in 98% of all PC for up to 120 h, and in 83% at 168 h. A swirling pattern score of 2.5 or greater predicted with a specificity of 100% a good morphology score in the PC. We conclude that PC prepared from buffy coats can be stored at 22 °C for up to 7 days. It is postulated that because the PC were leukocyte‐poor, the quality of the platelets was not influenced by the brand, or type of plastic in which they were stored.
Use of Polyester Filters for the Preparation of Leukocyte‐Poor Platelet Concentrates
Authors: Snyder Edward L. , DePalma Louis , Napychank Paula
Published: March 2009

Abstract. We studied the usefulness of two polyester leukocyte removal depth filters for the preparation of leukocyte‐poor platelet concentrate. The Sepacell R500A removed approximately 92% of the white cells and 98% of the platelets from stored concentrate. In contrast, the Travenol 4C2423 polyester depth microaggregate filter removed < 1 % of the platelets but only 14% of the leukocytes. These findings were obtained with stored (5‐day‐old) random donor platelets as well as with 16‐hour‐old platelets collected by apheresis. We conclude that neither of these two leukocyte removal depth filters are suitable for the preparation of leukocyte‐poor platelet concentrate; the Sepacell because it removes too many platelets and the 4C2423 because it removes too few leukocytes. The efficiency of cell removal by polyester filters is likely related to alteration of the polyester filter element during fiber manufacture.
The Transfused Population of Canterbury, New Zealand, and Its Mortality
Authors: Whyte Gordon S.
Published: March 2009

Abstract. All 367 recipients and 1,385 donations transfused over a 40‐day period were traced and compared by sex, age and discharge diagnosis with the 5,955 concurrent discharges from all general and obstetric hospitals in the area. 6% of discharged patients had been transfused. 21% of transfused patients were dead within 1 year, and at least 30% will die within 5 years. 228 donations of platelet concentrates were required for 7 recipients, 6 of whom died within 2 years. The mortality of blood recipients is overestimated if tracked from donors of multiple components.
Inactivation and Partition of Human Immunodeficiency Virus during Kistler and Nitschmann Fractionation of Human Blood Plasma1
Authors: Hénin Y. , Maréchal V. , Barré‐Sinoussi F. , Chermann J.‐C. , Morgenthaler J.‐J.
Published: March 2009

Abstract. We studied the inactivation of the etiologic agent of the acquired immunodeficiency syndrome, human immunodeficiency virus, in the course of the Kistler and Nitschmann cold ethanol fractionation of human blood plasma. By measuring reverse transcriptase activity and viral infectivity, we have shown that the virus load is reduced by a factor of 104 during the initial and at least a factor of 106 during the subsequent steps of the fractionation procedure. This loss of virus may be observed in the absence or in the presence of antibodies against human immunodeficiency virus and is due to a combination of chemical inactivation, physical partition, and injury caused by repeated freezing and thawing. The laboratory data therefore further confirm epidemiological studies which indicate that immunoglobulin preparations obtained by ethanol fractionation do not transmit human immunodeficiency virus.
Alloimmunization after Leukocyte‐Depleted Multiple Random Donor Platelet Transfusions
Authors: Brand A. , Claas F.H.J. , Voogt P.J. , Wasser M.N.J.M. , Eernisse J.G.
Published: March 2009

Abstract. In a prospective study we investigated the development and the course of alloimmunization after leukocyte‐depleted red cell and multiple random donor platelet transfusions in 335 patients. Of these 335 patients, who had a negative antibody screening on admission and a negative transfusion history, 69 (21%) developed either transien (n = 18) or permanent (n=51) lymphocytotoxic antibodies, but only 31 patients (9%; 95% confidence limits 6–12%) developed multispecific alloantibodies necessitating HLA‐matchedplatelet transfusions. There was no difference with regard to the development of antibodies and platelet refractoriness between leukemia patients receiving cytostatic treatment and patients with aplastic anemia receiving prednisone and antithymocyte globulin. Females with previous pregnancies developed platelet refractoriness with an increased incidence(x213.38; p < 0.001) compared to females without previous pregnancies, males, and children.
Studies on the Procurement of Blood Coagulation Factor VIII:
Authors: Griffin B. , Bell K. , Prowse C.
Published: March 2009

Abstract. To determine the viability of blood cells collected in half‐strength (0.5 CPD.A2) and full‐strength citrate anticoagulants, paired crossover autologous survival studies were performed in normal volunteers using 51Cr‐labelled red cells after 35 days storage and 111In‐labelled platelets after 5 days storage. For both studies, viability was better maintained in 0.5 CPD.A2 anticoagulant. This was significant for red cells (24‐hour survival: 80 ± 6 vs. 52 ± 21 %) but not for platelets by either linear or multiple‐hit models (recovery 51 ± 13 vs. 38 ± 14%, survival 5.1 ± 1.4 vs. 3.7 ± 1.5 days). The poor viability of red cells after storage in full‐strength anticoagulant was associated with low cellular adenosine triphosphate levels and was confirmed in follow‐up studies.
A Fatal Transfusion Reaction Associated with Blood Contaminated with Pseudomonas fluorescens
Authors: Scott J. , Boulton F.E. , Govan J.R.W. , Miles R.S. , McClelland D.B.L. , Prowse C.V.
Published: March 2009

Abstract. A fatal transfusion reaction due to contamination of platelet‐depleted whole blood with Pseudomonas fluorescens is reported. Routine sterility testing on blood products and environmental microbiological monitoring suggested no source for the contaminating organism, as has been the case for the majority of reported incidents of this type. The value of routine sterility testing in the prevention and investigation of such incidents is discussed.
Delayed Haemolytic Transfusion Reaction Due to Simultaneous Appearance of Anti‐Fya and Anti‐Fy5
Authors: Bowen D. T. , Devenish A. , Dalton J. , Hewitt Patricia E.
Published: March 2009

Abstract. We report here the development of anti‐Fy5 in a young Negro female with sickle cell disease. The antibody was responsible for a delayed haemolytic transfusion reaction. We believe this is the first report of such an antibody in Europe.
Large‐Scale Production and Properties of a Solvent‐Detergent‐Treated Factor IX Concentrate from Human Plasma
Authors: Michalski C. , Bal F. , Burnouf T. , Goudemand M.
Published: March 2009

Abstract. A human solvent‐detergent (SD)‐treated factor IX concentrate has been produced from cryoprecipitate‐poor plasma using DEAE‐Sepharose CL‐6B and heparin‐Sepharose CL‐6B chromatography. The DEAE eluate was incubated with an SD mixture [0.3% tri(n‐butyl) phosphate‐1% Tween 80, 6‐h at 24°C] which was found to inactivate, in less than 1 h, more than 3.8 logio of vesicular stomatitis virus and more than 4.8 logio of Sindbis virus; the SD was removed by a subsequent heparin adsorption step. The specific activity of the concentrate was 10.9 ± 1.3 IU factor IX: c/mg protein (n= 15). The factor IX coagulant to antigen ratio was 0.7 ± 0.1. The concentrate was essentially free of factors II, VII and X, and protein C. The usual major contaminants of prothrombin complex concentrate (PCC) were absent: the concentrate contained about 94% alpha‐1 proteins, and only 4 major proteins were resolved by SDS‐PAGE (respective apparent molecular weight: 130, 86, 76 and 69 kilodaltons), and by crossed immunoelectro‐phoresis against an anti‐PCC serum. The nonactivated partial thromboplastin time was equivalent to that of PCC; the product was devoid of factor IXa, of other activated procoagulant factors and of coagulant‐active phospholipids (removed with SD in the heparin breakthrough fraction). Animal studies using the Wessler test and acute‐toxicity test in rabbits revealed no adverse side effects. SD treatment could thus be used to inactivate viruses in factor IX concentrate and improve the safety of replacement therapy in hemophilia B.
The Bottom and Top System: A New Technique for Blood Component Preparation and Storage
Authors: Högman Claes F. , Eriksson Lars , Hedlund Kenneth , Wallvik Jonas
Published: March 2009

Abstract. A new, automated technique for the preparation of blood components is described. A system of 3 or 4 integrally connected plastic containers (Optipac®) is handled by a new type of extractor (Optipress®). The container in which the blood is collected has an outlet at the top and another at the bottom. After normal centrifugation to obtain separation of the blood components, these are squeezed out from the top and bottom simultaneously under control of a photocell. The primary separation step results in three components: a leukocyte‐poor red‐cell suspension in SAGM medium, CPD plasma, and a buffy‐coat preparation. The system has been tested in two laboratories (lab A and lab B). A ‘heavy‐spin’ centrifugation to obtain a maximum yield of cell‐poor plasma gave the best removal of leukocytes from the red cells; the remaining leukocyte content was 0.46 ± 0.25 (lab A) and 0.5 ± 0.4 (lab B)x 109/red‐cell unit. Platelet concentrates can be prepared either the normal way via platelet‐rich plasma or from buffy coat. Red‐cell 24‐hour autologous posttransfusion survival using labeling with 51Cr was 87.5 ± 4.1% (lab A) after 35 days, and 84.2 ± 4.2% (lab A) and 77.5 ± 1.5% (lab B) after 42 days. Red‐cell morphology and fluidity compared favorably to previous studies using the same additive solution in traditional plastic‐bag systems. The total adenine nucleotide concentration was maintained normal for 42 days. Storage hemolysis after 6 weeks was slightly higher after a heavy spin, 0.52 ± 0.19% (lab A) and 0.23 ± 0.10% (lab B), than after a light spin, 0.31 ± 0.15 and 0.20 ± 0.09%, respectively. The formation of fibrinopeptide A was very low during the first 2 weeks and then increased to 15 ± 15 nmol/1 after a heavy spin and to 33 ± 11 nmol/1 after a light spin. Kallikrein and spontaneous proteolytic activity increased from day 14 on in the light‐spun units but not in the heavy‐spun ones. Clinical studies were made in two hospitals. A total of 1,492 transfused blood components prepared with the new system were compared with 1,169 components prepared with a traditional four‐container system for making buffy‐coat poor red‐cell suspensions. No unexpected transfusion reactions occurred. The frequency of febrile reactions was 3/516 (new method) and 2/466 (controls) in a group of patients with predominantly myeloproliferative diseases. The system appears to be a significant improvement for automated preparation of high‐quality blood components.
Maternal‐Fetal Immune Relationship: Another Point of View
Authors: Vliet J. Husson‐van , Druyts‐Voets E.
Published: March 2009

Abstract. Placental transfer of maternal IgG is known to be an active process resulting in a concentration on the fetal side of the circulation ‐ at the apparent expense of that in the mother ‐ at full term. However, when this apparent decrease in maternal IgG is related to the actual hemodilution occurring in parallel, the total inventory of the mother's IgG appears to be constant throughout pregnancy. Therefore, conclusions regarding the variations in IgG concentrations cannot be drawn without simultaneous information about plasma volume fluctuations.
A New Platelet‐Specific Antigen, Naka, Involved in the Refractoriness of HLA‐Matched Platelet Transfusion
Authors: Ikeda Hisami , Mitani Takako , Ohnuma Miki , Haga Hiroshi , Ohtzuka Sayuri , Kato Toshiaki , Nakase Toshie , Sekiguchi Sadayoshi
Published: March 2009

Abstract. Serum from a thrombocytopenic patient who was refractory to the transfusions of HLA‐matched platelets contained a platelet‐specific alloantibody, anti‐Naka. Immunofluorescence analyses revealed that the Naka antigen defined by the serum was expressed exclusively on platelets and its distribution was different from PlA1, Baka, Yuka or Yukb. Analysis by Dr. von dem Borne's group revealed the Naka was also different from Koa, Kob or Zwb. Family studies showed that the Naka antigen was inherited as an autosomal codominant trait. Its antigen frequency in the Japanese population was over 97%. The results of the enzyme immunoassay using monoclonal antibodies for antigen immobilization showed that the Naka epitope did not appear to reside on GPIIb/IIIa or Ib. The transfusions of Naka‐compatible platelets improved the patient's thrombocytopenia.
Characterization of an IgG2, G2m(23) Anti‐Rh‐D Antibody
Authors: Dugoujon J. Michel , de Lange Gerda G. , Blancher Antoine , Alie‐Daram Simone , Marty Yvonne
Published: March 2009

Abstract. IgG2, G2m(23) anti‐Rh‐D antibodies were detected in the serum of a donor. It has been characterized by Gm typing and the use of polyclonal and mouse monoclonal anti‐IgG subclasses (which had proved to be satisfactory for agglutinaton of sensitized red blood cells). This anti‐D constitutes a good reagent for G2m(23) typing.
Identification, by Immunoblotting, of the Structures Carrying Lutheran and Para‐Lutheran Blood Group Antigens
Authors: Daniels Geoff , Khalid Ghizala
Published: March 2009

Abstract. Immunoblotting was performed, after sodium dodecyl sulphate polyacrylamide gel electrophoresis of solubilised red cell membranes under non‐reducing conditions, with human Lutheran antibodies, anti‐Lua, ‐Lub, ‐Lu3, ‐Lu6 and ‐Lu8, and with human para‐Lutheran antibodies, ‐Lu4, ‐Lu12 and ‐Lu17. On antigen‐positive red cells all antibodies revealed two components of apparent molecular weight (Mr) 83,000 and 76,000. Anti‐Lua and ‐Lub gave no bands with Lu(a‐b+) or Lu(a+b‐) cells, respectively. With the exception of anti‐Lu17, which reacted normally, and anti‐Lu3, which reacted weakly, none of the antibodies showed bands with membranes from Lu(a‐b‐) cells of the In(Lu) type. Treatment of the red cells with sialidase led to a small reduction in the apparent Mr of the Lutheran glycoproteins which, therefore, appear to contain sialic acid bearing O‐linked oligosaccharides.
IgG Subclasses of Anti‐A and Anti‐B Antibodies Bound to the Cord Red Cells in ABO Incompatible Pregnancies
Authors: Ukita M. , Takahashi A. , Nunotani T. , Kihana T. , Watanabe S. , Yamada N.
Published: March 2009

Abstract. IgG subclasses were determined in 138 A or B infants weighing over 2,500g, born to O mothers. Direct antiglobulin test (DAT) was positive in 43 infants and negative in 95 with anti‐A and/or anti‐B antibodies detected by heat elution test. In 59 out of 131 infants without ABO hemolytic disease (ABO‐HDN), no IgG subclass was detectable. In the 72 others, IgG1 was found in 29/72, IgG2 in 63/72, and IgG3 was not detected. In 7 infants with ABO‐HDN, DAT was positive in 4 and negative in 3. In conclusion, in DAT‐positive infants without HDN, IgG1 or IgG2 may be bound to erythrocytes, but the amount of IgG1 is too small to cause hemolysis. In DAT‐positive ABO‐HDN the amount of IgG1 is sufficient to cause hemolysis. In DAT‐negative ABO‐HDN, IgG3 is responsible for hemolysis, even though undetectable by DAT.
Cromer‐Related Antigens – Blood Group Determinants on Decay‐Accelerating Factor
Authors: Daniels Geoff
Published: March 2009

Abstract. Cromer‐related antigens, a series of blood group antigens phenotypically related to each other through various serological characteristics, have recently been shown to be carried on the complement regulatory glycoprotein decay‐accelerating factor (DAF). Cromer‐related antigens, therefore, represent a number of inherited variants and polymorphisms of DAF. The Inab phenotype, a ‘Cromer‐null’ phenotype in which the red cells lack all Cromer‐related antigens, appears to be an inherited DAF deficiency.
Quantification by ELISA of Erythrocyte‐Bound C3 Fragments Expressing C3d and/or C3c Epitopes in Patients with Factor I Deficiency and with Autoimmune Diseases
Authors: Rasmussen Jens Møller , Jepsen Hans Henrik , Teisner Børge , Holmskov‐Nielsen Uffe , Rasmussen Glen Gorm , Svehag Sven‐Erik
Published: March 2009

Abstract. A sensitive ELISA assay for quantifying erythrocyte (E) bound C3 fragments was developed. The assay employs a double‐antibody sandwich technique, using polyclonal anti‐C3d or anti‐C3c antibodies to quantify C3 fragments, expressing C3d and/or C3c epitopes in washed, detergent‐solubilized E. The assay detected 50–120 molecules of C3d per E in healthy individuals. Antigens reacting with anti‐C3c antibodies were also detected on E from normal individuals, but the density of C3c‐epitopes was 0.9–2.4 times lower than that of C3d‐epitopes. In 2 patients with congenital factor I deficiency significantly increased density of E‐bound C3c‐ as well as C3d‐antigen was observed. Plasma infusion in one of the patients induced a loss of E‐bound C3c‐antigens, indicating cleavage of E‐bound C3b to iC3b and further to C3c and E‐bound C3d. Loss of C3c‐antigens also occurred following in vitro treatment with normal human serum of E from one of the patients. Two thirds of 22 patients with systemic lupus erythematosus (SLE) and of 18 patients with rheumatoid arthritis had significantly increased density of E‐bound C3d, the highest density being 490 C3d molecules/E in an SLE patient. The density of E‐bound C3d correlated with the plasma‐C3d concentration, indicating that the coating of E with C3d reflects the degree of complement activation.
Ability of Monoclonal Anti‐D Antibodies to Promote the Binding of Red Cells to Lymphocytes, Granulocytes and Monocytes
Authors: Merry A.H. , Brojer E. , Zupanska B. , Hadley A.G. , Kumpel B.M. , Hughes‐Jones N.C.
Published: March 2009

Abstract. Red cells sensitized with IgG1 or IgG3 monoclonal anti‐D antibodies were used in rosette assays with human lymphocytes, monocytes and granulocytes. With all three cell types, IgG3 antibodies promoted a greater degree of rosette formation than IgG1 antibodies. Monocytes required a minimum of about 0.5 times 103 IgG3 molecules per red cell for rosette formation, and granulocytes and lymphocytes required around 1 times 104 IgG3 molecules per red cell. Approximately 80% of monocytes and granulocytes and 10% of lymphocytes were capable of rosette formation. These results are consistent with differences in the number and affinity of Fc receptors on different leucocytes. When compared with previous data these results suggest that binding of monocytes to monoclonal anti‐D sensitized red cells is very similar to that of red cells sensitized with polyclonal antisera. Lymphocytes and granulocytes, however, appear to bind less well to red cells sensitized with certain monoclonal antibodies than with polyclonal antibodies. These findings may be of relevance to the prophylactic use of monoclonal anti‐D antibodies.
Importance of a Quality Assurance Scheme for Factor VIII Assays in Quality Monitoring of Human Plasma Destined for Fractionation into Factor VIII Concentrate
Authors: Gabra G.S. , Prowse C.V. , Boulton F.E.
Published: March 2009

Abstract. A national quality assurance scheme has been established to monitor the validity of factor VIII assays performed by the various laboratories of the Scottish National Blood Transfusion Service engaged in collection and processing of donor plasma destined for fractionation. The results over the first 3‐year period show that comparable assay values can be obtained by participating centres using a common standard, despite differences in equipment, methods or substrate chosen for the one‐stage assay. The results also showed that chromogenic factor VIII assays correlated well with the one‐stage method. Random factor VIII assays performed on plasma, harvested and frozen within 18 h from collection, were analysed to validate recently proposed Scottish specifications which stipulate that 70% of plasma donations destined for fractionation should contain at least 0.7 IU/ml. Plasma harvested and frozen between 8 and 18 h from collection did not meet the specified level in any of the regional centres. This nationally specified level was also not met by plasma harvested and frozen within 8 h from collection in spite of being achieved individually by three regional centres. Assays performed on large plasma pools at the Fractionation Centre suggested loss of some factor VIII during storage, transportation and thawing of plasma prior to bulk processing.
Transfusion History, T Cell Subsets and Natural Killer Cytotoxicity in Patients with Colorectal Cancer1
Authors: Tartter Paul Ian , Steinberg Bryan , Barron Deborah M. , Martinelli Giorgio
Published: March 2009

Abstract. Blood transfusions are associated with clinical phenomena which are attributable to immune suppression. Since suppression of immune function is associated with a high risk of spontaneous malignancies, we studied T cell subsets and natural killer (NK) cytotoxicity in colorectal cancer patients and correlated the results with patients' transfusion histories. Twelve percent (14) of the 115 patients had been transfused an average of 19 years previously. Recipients of blood transfusions had low levels of peripheral lymphocytes (p=0.191), T cells (p=0.015), helper cells (p=0.016) and suppressor cells (p=0.2651) compared to previously untransfused patients. NK cytotoxicity was also significantly reduced in transfusion recipients although NK cell numbers were comparable in both groups. These results support previous studies indicating that blood transfusions cause lifelong immune modulation in the recipient. Since blood transfusions have numerous beneficial effects and immune modulation is often beneficial, longitudinal studies are necessary to define the lifetime risks and benefits.
Storage of Leukocyte‐Poor Red Cell Concentrates: Filtration in a Closed System Using a Sterile Connection Device
Authors: Pietersz R.N.I. , Reesink H. W. , de Korte D. , Dekker W.J.A. , van den Ende A. , Loos J.A.
Published: March 2009

Abstract. Storage of leukocyte‐poor red cell concentrates (LP‐RCC) was investigated after filtration in a closed system that was assembled using a Sterile Connection Device (SCD). The LP‐RCC were stored for up to 6 weeks following filtration with either 0.9% saline solution (n = 14) or saline‐adenine‐glucose‐mannitol (SAG M) solution (n = 15) to prime and rinse the cellulose acetate filter. The results were compared with the data of nonfiltered buffy‐coat‐poor red cell concentrates (BC‐poor RCC) stored in SAG M solution (n = 10). All LP‐RCC contained less than 106 leukocytes whereas the nonfiltered BC‐poor RCC contained 6751286 × 106 leukocytes at day 1, decreasing to 83±49 × 106 at day 42. Although glucose consumption, lactic acid production and decrease in pH was similar from day 7 through 28 in both groups of LP‐RCC, a significantly steeper decline of ATP values as well as a higher hemolysis and LDH release was observed in the LP‐RCC filtered with saline. During storage of the nonfiltered BC‐poor RCC in SAG M, significantly higher glucose consumption (p<0.01), LDH release (p<0.001), rate of hemolysis (p<0.001) and a lower pH (p<0.001) were found, compared to the filtered units. It is postulated that the leukocytes present in the nonfiltered BC‐poor RCC were responsible for these differences. The ATP values in the SAG‐M‐filtered and nonfiltered BC‐poor RCC in SAG M were comparable. By comparing the ATP levels and values of the filtered RCC and the nonfiltered BC‐poor RCC we conclude that the LP‐RCC can be stored for 35 days if SAG M solution is used to prime and rinse the filter.
Donath‐Landsteiner Autoimmune Hemolytic Anemia in Children
Authors: Göttsche B. , Salama A. , Mueller‐Eckhardt C.
Published: March 2009

Abstract. During the last 4 years, we have studied a total of 531 adults and 68 children with clinically and serologically well‐defined forms of immune hemolytic anemias. Among these, Donath‐Landsteiner (DL) hemolysis was the underlying disease in 22 of the 68 children (32.4%), but was not observed in adults. All children with DL hemolysis suffered from acute infections presumably of viral origin. In none of the cases was the DL hemolysis suspected clinically. Boys were more often affected than girls. The hemolytic episodes were severe, but resolved within few weeks. Serologically, all patients had a strongly positive direct antiglobulin test (DAT) using anti‐C3d reagents, but a weak (n = 6) or negative (n = 16) IgG‐DAT. DL hemolysins were always weak and transient, detectable with enzyme‐treated red blood cells (RBC) in all, with untreated RBC in 12 of 22 sera. To explore the reason why these weak antibodies can cause extensive hemolysis in vivo, we compared the action of DL antibodies and of cold agglutinins (anti‐I) on RBC by several reincubations at 4 and at 37°C. The data obtained from this experiment demonstrate a stronger aggravation of hemolysis by DL than by anti‐I antibodies, presumably due to low‐affinity interaction between the cells and DL antibodies.
Abnormal Beta and Gamma Sialoglycoprotein Associated with the Low‐Frequency Antigen Lsa
Authors: Macdonald E. B. , Condon J. , Ford D. , Fisher B. , Gems L. M.
Published: March 2009

Abstract. This paper describes the association of the low‐frequency antigen Lsa with high‐molecular‐weight variants of β‐ and γ‐sialoglycoproteins (β‐SGP, γ‐SGP). The variant β‐SGP, designated βTLa, carries the Ge3 epitope and the epitopes recognised by 3 murine monoclonal anti‐β‐SGP antibodies. The variant γ‐SGP, designated γLsa, carries the Ge2 and Ge3 epitopes. Both βLsa and γLsa showed enhanced reactions in immunoblotting with anti‐Ge3 sera — which may indicate the presence of 2 Ge3 epitopes on (βLsa and γLsa. These findings would be consistent with the proposed structure of the Lsa glycophorin C gene.
The Impact of Preoperative Autologous Blood Donation on Orthopaedic Surgical Practice
Authors: Goodnough L. T. , Shafron D. , Marcus R. E.
Published: March 2009

Abstract. We have conducted a retrospective 3‐year analysis of our autologous blood donation program to assess its impact on orthopaedic surgery. We conclude: (1) utilization has increased from less than 5% of eligible patients in the first audit interval to nearly 50% in the last audit interval; (2) in the last audit interval, autologous blood donation resulted in a reduction of homologous blood transfusion from 41% in nonautologous blood patients to 14% in autologous blood donors; (3) increasingly conservative transfusion practice is seen for all patients undergoing elective orthopaedic surgery; (4) regional blood centers are responding to increasing requests for autologous blood with programs that are effective in attracting autologous blood donors; (5) on the basis of utilization and efficacy, preoperative autologous blood donation as an alternative to homologous blood transfusion now represents a standard of practice for elective orthopaedic surgery.
Multiple Epitopes Recognised by Human Monoclonal IgM Anti‐D Antibodies
Authors: Hughes‐Jones N.C. , Gorick B.D. , Beale D.
Published: March 2009

Abstract. Bivalent 7S subunits were obtained from 7 purified IgM human monoclonal anti‐D antibodies with 33–63% of the total protein‐retaining functional binding activity. The number of sites per red cell recognized by these subunits ranged between 9,400 and 28,500. The values are taken to indicate that a number of different epitopes on the D polypeptide are being recognized. There was competition between IgG and IgM antibodies for binding to the red cells, indicating that both classes of antibody are recognizing epitopes on the same D polypeptide. The value of the functional affinity constants for the binding of the 7S subunits varied between 1.0 and 8.8 times 107 M‐1 and there was a 3‐ to 16‐fold increase on reduction of the ionic strength (0.05 M NaCl). Using agglutination in microwells, the end‐point of the titres for the native pentamer IgM antibodies occurred at concentrations in the range 7–26 ng/ml, with one exception where the concentration required was 165 ng/ml.
Non‐A, Non‐B Hepatitis and the Anti‐HCV Assay
Authors: Barbara John A. J. , Contreras Marcela
Published: March 2009

Abstract. The successful cloning of a non‐structural antigen from the genome of what is now designated as the ‘hepatitis C virus’ (HCV) has transformed an erstwhile diagnosis of exclusion for non‐A, non‐B hepatitis (NANBH). The assay has been validated against panels of known infectivity for NANBH and sera from haemophiliac patients treated either with virally inactivated or uninactivated factor VIII. The predictive value of the assay is being assessed clinically in prospective studies of post‐transfusion hepatitis and by using laboratory techniques such as polymerase chain reaction. While the assay shows good predictability in high‐risk subjects, an appreciable number of false‐positive results are likely in blood donor populations. Furthermore, the extent of infectivity of seropositive blood donors is still the subject of active research. The prevalence of anti‐HCV in blood donors varies from approximately 0.2 to 1.5% around the world, based on repeat reactivity in the Ortho antiglobulin ELISA assay. These rates may be appreciably reduced following supplementary testing with recombinant immunoblot assay (RIBA). Prevalence data in African sera are as yet unreliable, pending assessment by RIBA, presumably because of high levels of IgG interfering with the assay. Presence of anti‐HBc or elevated alanine aminotransferase associates to a greater or lesser extent with seropositivity, especially when both surrogate markers are present, but conversely many (unconfirmed) seropositive subjects lack these surrogate markers. An understanding of the modes of transmission of HVC is of obvious importance to transfusion practice. Intravenous drug use is a striking risk factor, but the contribution made by sexual transmission is not so clear. Results with the highly innovative anti‐HCV assay represent the first dramatic ‘ranging shots’ to hit the target of NANBH, albeit at the periphery. Some of the various limitations of this expensive assay and associated supplementary tests are likely to be resolved by the advent of improved tests based on other HCV antigens, including structural ones. Hopefully, they will produce results closer to the centre of the target.