bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Microfiltration on the Histamine Levels in Stored Human Blood
Authors: Frewin D.B. , Jonsson J.R. , Davis K.G. , Beilby A.M. , Haylock D.N. , Beal R.W. , Russell W.J.
Published: March 2009

Abstract. Plasma and whole blood histamine levels and white cell counts have been monitored over a 6‐week period of storage in standard citrate‐phosphate‐dextrose supplemented with adenine (CPDA) blood packs, add‐back blood packs, and also CPDA and add‐back packs which were subjected to a microfiltration process at the time of collection. Our results indicate that the plasma histamine level rises and the white blood count falls progressively with time, the former being most marked after the 3rd week of blood storage. At 6 weeks, there was no significant difference between the plasma and whole blood histamine level, suggesting that the entire complement of histamine in blood was now in plasma. Microfiltration substantially reduced the white cell count in the packs, and this was reflected in a greatly reduced whole blood and (subsequently) plasma histamine concentration.
Fatal Immune Hemolytic Anemia and Hepatic Failure Associated with a Warm‐Reacting IgM Autoantibody
Authors: Shirey R. Sue , Kickler Thomas S. , Bell William , Little Barbara , Smith Barbara , Ness Paul M.
Published: March 2009

Abstract. Autoimmune hemolytic anemia (AIHA) caused by warm‐reacting IgM autoantibodies is rare. We report a fatal case of primary AIHA with a warm‐reacting IgM autoantibody. Recurrent episodes of intravascular hemolysis, unresponsive to all therapy and progressive hepatic dysfunction characterized the patient's clinical course. Despite corticosteroid therapy, splenectomy and multiple blood transfusions, the patient died from liver failure. The IgM autoantibody caused autoagglutination of the patient's red cells at 37 °C. Eluates prepared from the patient's red cells agglutinated saline‐suspended test cells without the addition of antiglobulin reagent. We propose that warm‐reacting IgM antibodies may lead to in vivo autoagglutination and may be associated with hepatic failure.
Study of Anti‐Fya in Five Black Fy(a‐b‐) Patients
Authors: Le Pennec P. Y. , Rouger Ph. , Klein M. T. , Robert N. , Salmon Ch.
Published: March 2009

Abstract. Fya antibodies were detected in 5 Fy(a‐b‐) black patients during a 5‐year period. The Duffy phenotype was confirmed using anti‐Fya, anti‐Fyb, and anti‐Fy3 by an adsorption‐elution technique. These 5 patients had sickle cell disease and had received multiple blood transfusions. The antibodies were all found during the investigation of a hemolytic transfusion reaction or failure to achieve the desired red cell increments after transfusion.
Studies on the Procurement of Blood Coagulation Factor VIII in vitro Studies on Blood Components Prepared in Half‐Strength Citrate Anticoagulant
Authors: Prowse C. , Waterston Y. G. , Dawes J. , Farrugia A.
Published: March 2009

Abstract. The effect of replacing a standard citrate anticoagulant with one containing half the amount of citrate on the in vitro properties of components prepared from blood donations was investigated. This resulted in a significant improvement in factor VIII stability such that there was little loss during overnight storage, and this was reflected in the factor VIII yield in cryoprecipitate. The quality of cellular components in red cell units stored up to 35 days or platelet concentrates stored up to 7 days was not adversely affected. Although initial levels were similar to those in standard anticoagulant, the extent of fibrinopeptide A generation and complement C3 breakdown in red cell units stored for 35 days in half‐strength citrate was somewhat increased.
Effects of Plasticizers and Plastic Bags on Granulocyte Function during Storage
Authors: Miyamoto Masaki , Sasakawa Shigeru
Published: March 2009

Abstract. The influence of the plasticizers, di‐(2‐ethylhexyl)phthalate (DEHP) and tri‐(2‐ethylhexyl)trimellitate (TOTM), on granulocyte function was examined. Polyvinyl chloride (PVC) bags with DEHP (DEHP‐PVC) leaked DEHP into plasma, but TOTM did not dissolve in plasma under the same conditions. Glow discharge treatment inhibited the leakage of DEHP from DEHP‐PVC bags. Depending on the amount of DEHP added into granulocyte suspension, chemotaxis and bactericidal activity decreased, but cell counts and phagocytosis were not affected. During storage for 24 h at 22 °C, granulocyte function decreased greatly in DEHP‐PVC, but was well maintained in the bags which did not leak plasticizers, TOTM‐PVC and glow‐discharged DEHP‐PVC.
Monitoring of Platelet Contamination in Filtered Red Blood Cell Concentrates
Authors: Vos J.J.E. , Schoen C. , Prins H.K. , von dem Borne A.E.G.Kr. , Huisman J.G.
Published: March 2009

Abstract. Monitoring of contaminating platelets, granulocytes, and lymphocytes in leukocyte‐poor red blood cell concentrates is usually done by counting in an electronic particle counter. Sensitivity and specificity of this technique are compromised by the contamination of the preparations with other cell types and particles thereof. In this report we studied platelet contamination in filtered red blood cell concentrates by use of a radioimmunoassay for detection of the platelet glycoprotein complex Ilb‐IIIa. Our results indicate that platelets and/or fragments thereof, not detectable for particle counters, are present in blood cell concentrates. This finding might have important implications for the preparation of pure red blood cell concentrates to avoid unwanted immunization after transfusion.
Higher ATP Level and Reduced Glucose Consumption of Red Cells Stored in PVC Bags Compared to Glass Bottles
Authors: Hegedüs Edit , Rácz Zoltdn
Published: March 2009

Abstract. Whole blood stored for 21 days at +4 °C in glass bottles showed a decrease in plasma glucose content of 6.6 mM/1, while blood stored in polyvinylchloride bags had a decrease of only 5.3 mM/1. At the same time, red cell ATP was reduced to 69% in bottles compared to 80% in bags (statistically proved differences). The higher red cell ATP level may explain the increased stability of red cells stored in DEHP‐plasticized blood bags. The reduction in glucose consumption suggested that the decrease of red cell ATP and plasma glucose content may be a result of some unidentified inhibitory effects of DEHP on biochemical processes.
Phagocytosis of Erythrocytes Sensitized with Known Amounts of IgGl and IgG3 Anti‐Rh Antibodies
Authors: Zupańska B. , Thompson E. , Brojer E. , Merry A.H.
Published: March 2009

Abstract. Phagocytosis was investigated using human peripheral monocytes and erythrocytes sensitized with known amounts of subclass‐specific IgG anti‐Rh antibodies. The erythrocyte‐bound IgG was quantitated by a radiometric antiglobulin test. This evaluation revealed the following: (1) there is a relationship between phagocytosis and the number of erythrocyte‐bound IgG molecules; (2) phagocytosis is IgG subclass‐dependent, since a similar degree of phagocytosis is observed with fewer IgG3 than IgGl molecules and also the minimum number of IgG3 molecules for phagocytosis is 150–640, whilst for IgGl the minimum is 1,230‐4,020; (3) the minimum levels of sensitization for phagocytosis should be detectable by the serological antiglobuiin test; (4) the phagocytosis assay is no more sensitive than the monocyte rosette assay for the detection of anti‐Rh alloantibodies, and (5) phagocytosis of adherent erythrocytes observed by video‐enhanced microscopy indicated that erythrocytes may adhere to monocytes for a considerable time before phagocytosis, but that phagocytosis itself was rapid.
Apparent ‘Auto’‐Anti‐DE of the IgA and IgG Classes in a 16‐Year‐Old Girl with a Mature Cystic Ovarian Teratoma
Authors: Schönitzer D. , Kilga‐Nogler S.
Published: March 2009

Abstract. Prior to the removal of a mature cystic teratoma in a girl, aged 16, we had difficulties in Rh typing due to immunoglobulin coating of the red blood cells (RBCs). Further examination revealed the presence of anti‐D and anti‐E antibodies; the clinical course and the serological characteristics indicated that the Rh antibodies originated from the tumor itself, that is, there was alloantibody formation against the host's RBCs. The specific alloantibody production, as well as the fact that these antibodies belong to the IgA and IgGl classes provide further evidence for the pluripotency of mature cystic ovarian teratomas.
Assessment of a New Centrifugal Plasmapheresis Machine: The SPC‐600
Authors: Prowse C. V. , Waterston Y. , Dawes J. , McClelland D.B.L. , Gillon J.
Published: March 2009

Abstract. An approach to the full assessment of new plasmapheresis machines is outlined. This involves testing for changes in donated blood, donors and blood components using fresh donor blood and during full procedures in an animal model prior to performing full procedures in human volunteers. Using this approach, a new centrifugal plasmapheresis machine, the SPC‐600, was assessed. No adverse effects associated with the use of the machine were detected in the initial studies. When used for plasmapheresis of volunteers, 500 ml of plasma was obtained in 43 ± 4 min without adverse effects on donors. The plasma product contained normal levels of plasma proteins, including factor VIII, and 20 ± 6× 109/1 platelets. There was no evidence of activation of complement or coagulation systems, and the factor VIII yield in cryoprecipitate prepared from single donations of plasma was equivalent to that observed for standard whole blood donations.
Use of Fab Anti‐IgG in Phenotyping IgG‐Sensitised Red Cells
Authors: Henry S.M. , Baird S.J. , Woodfield D.G.
Published: March 2009

Abstract. Previously described methods of phenotyping red cells sensitised with IgG using the indirect antiglobulin test required the dissociation of the coating protein. Based on an entirely different principle, Fab fragments of anti‐human IgG (Fab anti‐IgG) were used to block the antiglobulin binding sites on cell‐bound IgG molecules, removing the necessity to dissociate them from the red cell. Fab anti‐IgG was found to be effective in blocking interfering IgG on in vivo and in vitro IgG‐sensitised red cells, permitting successful red cell phenotyping. Strongly IgG‐sensitised samples which could not be fully neutralised by chloroquine diphosphate (CDP) or blocked with Fab anti‐IgG alone could usually be phenotyped using a combination of both these methods. This new procedure may be of use in immunohaematology laboratories.
Mimicking Anti‐S Simulating a Delayed Transfusion Reaction
Authors: Puig N. , Carbonell F. , Soler M.A. , Perez M.T. , Arriaga F. , Marty M.L.
Published: March 2009

Abstract. An autoantibody mimicking anti‐S specificity is described in an S‐s+ patient. The antibody was detected 2 weeks after transfusion. The direct antiglobulin test (DAT) was positive and the antibody was thought to be an alloantibody formed after the transfusion. Eighteen months later, the DAT was still positive and anti‐S could be eluted from the patients's red blood cells in spite of the fact that these were S‐s+. The autoantibody was characterized as mimicking anti‐S specificity by in vitro absorption studies. It was present during the 3 years in which the patient was observed without causing hemolitic anemia.
Survival of Red Cells Stored for 21 and 35 Days in a Non‐Di‐(2‐Ethylhexyl)Phthalate Plastic Container
Authors: Myhre Byron A. , Johnson Delores , Marcus Carol S. , Demaniew Suzanne , Carmen Raleigh , Nelson Ed
Published: March 2009

Abstract. Whole blood and red cells were stored using citrate‐phosphate‐dextrose (CPD) and citrate‐phosphate‐dextrose‐adenine (CPDA‐1) anticoagulants in polyvinylchloride bags made flexible with di‐(2‐ethylhexyl)phthalate (DEHP) or tri‐(2‐ethylhexyl)trimellitate (TOTM) plasticizers. After storage the posttransfusion viability of these cells was tested in autologous donors. Cells stored in TOTM‐plasticized film had a survival rate less than 75% when stored for 35 days, while other systems had a survival greater than this. When compared with the red cells stored in CPD‐DEHP‐plasticized film, the viability of whole blood and red cells stored in CPDA‐TOTM showed a statistically significant decrease (p = <0.01). Therefore, red cell storage in TOTM‐plasticized PVC with current anticoagulant should be limited to 21 days.
Preparation of Leukocyte‐Poor Platelet Concentrates from Buffy Coats
Authors: Pietersz R.N.I. , Reesink H. W. , Dekker W.J.A. , Fijen F.J.
Published: March 2009

Abstract. A special insert was developed for centrifuge cups in order to prepare leukocyte‐poor platelet concentrates from buffy coats by using quadruple citrate phosphate dextrose‐saline adenine glucose mannitol systems from different manufacturers. Each centrifuge cup could contain up to 4 sets of double bags allowing the preparation of 24 platelet concentrates per run. Optimal conditions for centrifugation of the buffy coats in the inserts were found to be 6 min at 380 g (2,150 g min). A platelet count of 69 ± 19 × 109 and a leukocyte contamination of 14± 10.5 × 106 per platelet concentrate was thereby obtained in a plasma volume of 63 ± 10.5 ml (mean ±SD). The method described allows large scale production of leukocyte‐poor platelet concentrates from buffy coats in a closed system.
Platelet Storage Lesion in Second‐Generation Containers: Correlation with Platelet ATP Levels
Authors: Holme S. , Heaton W.A. , Courtright M.
Published: March 2009

Abstract. The nature of platelet lesion occurring with storage of platelet concentrates (PC) in second‐generation containers was investigated using various storage media and storage periods up to 14 days. In CPD‐plasma (control medium), the changes which occurred progressively during storage were loss of discoid shape, microscopic platelet aggregate formation, fragmentation and the appearance of disintegrated, ‘balloon’ forms. By day 14 less than 10% of the platelets were discoid in shape, the platelet count had decreased by 23%, and there was a 5‐fold increase in the amount of lactate dehydrogenase in plasma. Associated with this was a decrease in the platelet oxygen consumption rate, D(O2), loss of cellular ATP and extent of ADP‐induced shape change, and a decrease in the hypotonic shock response. These parameters decreased at a similar rate, with a 50% decrease (t1/2) at days 7–9. They correlated highly with each other during storage and also with a fall in pH. At day 14 of storage, mean pH was 6.1 ± 0.3. To evaluate the effect of pH stabilization during storage, 4 mEq sodium bicarbonate was added to PC in CPD‐plasma. Although pH maintenance was much improved, 7.2–6.6 during 14 days of storage, the same in vitro lesions developed, although more slowly. The t1/2 of the same parameters was prolonged for approximately two days. When PC were stored in a plasma‐free physiologic salt solution whose salt composition was similar to CPD‐plasma, the t1/2 of the parameters increased to 11–15 days of storage, although the platelets eventually developed the same in vitro lesions. In all media there was a high correlation between platelet ATP level and the other in vitro parameters during storage. Furthermore, energy depletion of the platelets produced by using the physiologic salt solution without glucose and by addition of metabolic inhibitors caused a similar deterioration of in vitro functions, but at a significantly faster rate which correlated with the rate of decrease in platelet ATP. This suggests that the platelet lesion observed with prolonged storage in second‐generation containers might be due to a failure in platelet energy metabolism.
A Rapid and Accurate Single‐Drop Modification of the Acid‐Elution Technique for Detecting Fetomaternal Hemorrhage
Authors: Corsetti J.P. , Cowles J.W. , Cox M.T. , Blumberg N.
Published: March 2009

Abstract. A single‐drop modification of the acid‐eltion technique (Kleihauer‐Betke) for quantitating fetomaternal hemorrhage is described. It obviates the need for the tedious and time‐consuming manual counting of background adult cells. Rather, this is achieved by automated red‐blood cell counting of the initial specimen and delivery of a standard volume (1 μl) of a standard dilution (1:1,000) in the form of a droplet to a microscope slide. The droplet is left to dry undisturbed at room temperature and then stained. The fetal cells are manually counted while the total number of cells is calculated from the initial red‐blood cell count, standard volume, and standard dilution. Determinations on 4 different concentrations of fetal/adult red cell mixtures are performed. Results indicate improved accuracy and precision relative to the standard technique in significantly less time for volumes of fetomaternal hemorrhage requiring more than the standard dose of RhoD)‐immune globulin.
Considerations in the Use of Laboratory Robots: Aspects of Safety and Accuracy1
Authors: Brennan J.E. , Severns M.L. , Kline L.M. , Epley K.M.
Published: March 2009

Abstract. A general‐purpose laboratory robot was configured to prepare blood samples for the HBsAg and anti‐HIV tests. Several issues were studied in order to eliminate problems and optimize the system. These included pipetting accuracy, intersample carryover, reagent interference, splashing and aerosol generation and user‐related issues. A number of general guidelines were developed to assist others in developing and implementing systems for this and similar applications.
Efficacy of an Immunoglobulin Preparation from HIV Carriers in Preventing HIV Replication in vitro
Authors: Tochikura Tadafumi S. , Nakashima Hideki , Uemura Yahim , Goto Takashi , Suyama Tadakazu , Kobayashi Nobuyuki , Yamamoto Naoki
Published: March 2009

Abstract. Immunoglobulin samples (HIV‐Ig) were prepared by cold ethanol fractionation of human plasma containing antibody against human immunodeficiency virus (HIV). The ability to prevent viral spreading was studied using either human T‐cell leukemia virus type I (HTLV‐I)‐carrying MT‐4 cells or in a coculture system using MOLT‐4 cells and virus‐producing MOLT‐4/HIVHTLV‐IIIB cells. Treatment of HIV‐infected MT‐4 cells with HIV‐Ig effectively blocked the appearance of antigens of HIV and the virus‐induced cytopathic effect. HIV‐Ig blocked multinucleated giant cell formation in the MOLT‐4 and MOLT‐4/HIVHTLV‐IIB coculture system.
Studies on the Procurement of Blood Coagulation Factor VIII
Authors: Griffin B. , Bell K. , Prowse C.
Published: March 2009

Abstract. Our previous studies have shown that the use of half‐strength citrate anticoagulant improves plasma factor VIII stability while maintaining the viability of red cells and platelets. This study extends these observations to show that the stability of factor VIII is maintained in blood stored at room temperature or 4°C overnight and that in vitro tests indicate comparable quality of red cells and platelets prepared from half‐strength citrate donations processed immediately or after overnight storage at room temperature. The exception to this was a more rapid loss of red cell 2, 3‐diphosphoglycerate, as has previously been observed for full‐strength citrate donations.
Management of Alloimmune Neonatal and Antenatal Thrombocytopenia
Authors: Reznikoff‐Etievant M.F.
Published: March 2009

Abstract. Neonatal and antenatal alloimmune thrombocytopenia is induced by maternal antibodies against platelet‐specific fetal antigens. This disease is rare but potentially severe because of intracranial bleedings which may occur during pregnancy or around birth. In the last decade our knowledge of this disorder has markedly advanced. New techniques are used in platelet immunology. New platelet antigens involved in these perinatal thrombocytope‐nias have recently been discovered. A group of women likely to produce the responsible platelet antibodies has been genetically defined as regards the PLA1 antigen. The quality of the sonographies and the possibility of performing cord vein puncture in early pregnancy afford a new approach in the management of perinatal alloimmune thrombo‐cytopenias. But more must be done to prevent the complications of this disease.
Preparation of Leucocyte‐Poor Platelet Concentrates from Buffy Coats
Authors: Pietersz R.N.I. , Reesink H. W. , Huijgens P.C. , van Oers M.H.J.
Published: March 2009

Abstract. To study survival and function of leukocyte‐poor platelet concentrates (lp‐PC) prepared from buffy coats, random platelet transfusions requested for thrombocytopenic patients were evaluated. The lp‐PC issued were stored at 22°C for either 1, 3 or 5 days before transfusion. From 31 transfusions, posttransfusion corrected count increments (CCI), corrected for body surface (m2) and divided by number of platelets transfused (1011), were calculated. The mean ±SEM of the 1‐, 24‐and 48‐hour CCI was 12.2 ± 0.45, 11.2 ± 0.51 and 8.8 ± 0.58, respectively. No significant differences in CCI 24 h after transfusion were observed for lp‐PC stored for 1, 3 or 5 days. Hemostatic activity was observed in all 9 evaluable patients. It is concluded that platelets from lp‐PC survive well in patients, regardless of storage for 1, 3 or 5 days and that the platelets are hemostatically active after transfusion.
Distribution of Blood Gases, Glucose and Lactate within Stored Blood Units
Authors: Sandhagen Bo , Högman Claes F. , de Verdier Carl‐Henric , Eriksson Lars
Published: March 2009

Abstract. The distribution of respiratory gases, hydrogen ions, glucose and lactate were studied in red cells suspended in SAGM solution and stored undisturbed at +4°C in plastic (PVC) bags. After 42 days' storage there were considerable concentration differences between the supernatant and the red cell layer, as well as concentration gradients within the latter. Thus, pO2, sO2, pH and the glucose concentration were higher in the supernatnant and pCO2 and the lactate concentration were higher in the red cell layer.

These findings show that sampling from the mixed content of blood which has been stored sedimented in plastic containers will not be representative for the erythrocyte environment at the end of shelf life. This is of importance in attempts to understand storage effects on red cells.
Screening of Blood Donors for Antibody to Human Immunodeficiency Virus Type I by Sensitive Particle Agglutination Assay
Authors: Ohya K. , Morishima Y. , Funato E. , Hasegawa I. , Kamiya T.
Published: March 2009

Abstract. A particle agglutination (PA) assay for antibody to human immunodeficiency virus type I (anti‐HIV) was used to screen blood donors and to test leukemia and hemophilia patients. Results by PA showed complete agreement with the results of two kinds of enzyme immunoassay (EIA), and the false‐positive rate was the same as or lower than with the EIAs. The sensitivity of PA was 10‐ to 100‐fold higher than that of the EIAs. This simple sensitive assay takes less time and fewer personnel than the EIAs, and has been used for massive screening of blood donors in our blood center.
The Effect of 2 Molar Urea on Jk (a‐b‐) Red Cells
Authors: Edwards‐Moulds Jo Ann , Kasschau Margaret R.
Published: March 2009

Abstract. The phenomenon of red cell lysis in urea was investigated for both normal and Jk(a‐b‐) red cells. Using 14C urea, urea was shown to be transported across the membrane in both cell types. In normal cells this resulted in an influx of water, cell swelling and complete lysis within 2 min. However, in Jk(a‐b‐) red cells, the presence of urea above 300 mM slowed water movement and delayed hemolysis for at least 15 min. Similar effects could be induced by pretreating normal red cells with p‐chloromercuribenzene sulfonate (PCMBS) or by raising the pH of the urea solution to 8.0.
The 24‐Hour Posttransfusion Survival and Lifespan of Autologous Baboon Red Cells Treated with Inositol Hexaphosphate‐Polyethylene Glycol or Inositol Hexaphosphate‐Adenosine Triphosphate‐Polyethylene Glycol to Decrease Oxygen Affinity1
Authors: Franco R.S. , Weiner M. , Wagner K. , Martelo O.J. , Ragno G. , Pivacek L.E. , Valeri C.R.
Published: March 2009

Abstract.Baboon red cells were treated to reduce oxygen affinity by an osmotic‐pulse procedure using dimethyl‐sulfoxide. Inositol hexaphosphate (IHP) alone or IHP and adenosine triphosphate (ATP) were incorporated into the red cells in the presence of polyethylene glycol (PEG). The procedure produced variable increases in the red cell P50value, i.e., the partial pressure of oxygen at which 50% of the hemoglobin was saturated. The effect of treatment of autologous baboon red cells on the 24‐hour posttransfusion survival value and lifespan T50value was measured using a double‐label procedure. The data demonstrate that the increase in the P50value of treated red cells was negatively correlated with the 24‐hour posttransfusion survival value; the higher the P50value, the poorer the 24‐hour posttransfusion survival value. The 24‐hour posttransfusion survival value for nontreated baboon red cells was 90% and the T50value was 14 days. The IHP‐ATP‐PEG‐treated red cells had significantly higher red cell ATP levels than did IHP‐PEG‐treated red cells. The 24‐hour posttransfusion survival value was 68% for the IHP‐ATP‐PEG treated red cells and 52% for the IHP‐PEG‐treated red cells when the increase in P50ranged from 10 to 20 mm Hg; the lifespan T50value for both the IHP‐ATP‐PEG‐treated red cells and the IHP‐PEG‐treated red cells was 15 days. Osmotic pulse treatment produced significant red cell injury manifested by the 24‐hour posttransfusion survival value. However, modification of the RBC with IHP‐PEG or IHP‐ATP‐PEG to decrease hemoglobin affinity for oxygen did not affect their lifespan.
Quantitative Determination of Residual Sodium‐Cholate in Tri(N‐Buthyl)Phosphate/Sodium‐Cholate Treated Factor VIII‐Preparations
Authors: Dengler T. , Kellner S. , Fürst G.
Published: March 2009

Abstract. Abstract. A relatively simple and fast method for the quantitative determination of residual sodium‐cholate in tri(n‐buthyl)phosphate/sodium‐cholate treated factor VI concentrate preparations was developed by solid‐phase extraction and reversed‐phase high‐performance liquid chromatography.
Exclusion of Unassigned Blood Group System Loci from the Regulator of Complement Activation Gene Cluster on Chromosome 1q32
Authors: Lewis M. , Kaita H. , Philipps S. , McAlpine P.J. , Wong P.
Published: March 2009

Abstract. Genetic linkage analyses of the blood group system loci CO, DI, DO, KEL and YT in relation to F13B indicate that these loci are not members of the RCA gene cluster on chromosome 1q32. The data are presented to finalize the exclusion of the DAF carried red cell antigens from all of the 17 established blood group systems.
A Semi‐Continuous Method for Purification of Factor IX Complex from Human Plasma
Authors: Tharakan John P. , Gee Dan M. , Clark David B.
Published: March 2009

Abstract.We report the development of a semi‐continuous method for preparation of factor IX complex from human plasma using ion exchange resins. Traditionally, stirred batch adsorption has been used due to the high pressure drops and low flow rates associated with soft gels in packed columns. Batch methods, however, typically involve higher labor costs and are more cumbersome in process environments. In the semi‐continuous process, cryo‐supernatant plasma is pumped through a ‘stirred column’ containing the resin. At both lab and pilot scale, higher recoveries of factor IX (FIX) were obtained at decreased total process times, compared to batch adsorption. A residence time of 15 min was found to be sufficient for capture of 95% of the FIX in the starting plasma. In the pilot plant, 550 liters of plasma was passed through a 50‐liter column containing 8.5 liters of resin, yielding a 68% recovery of FIX. The results suggest that the recovery of FIX depends on the mode of contact (batch, continuous or packed column) between plasma and resin.
The Role of Antibody Density in the Immune Lysis of Sensitised Erythrocytes: A Mathematical Appreciation
Authors: Bakács Tibor , Ringwald Gábor , Léránth Tünde , Kimber Ian
Published: March 2009

Abstract. The influence of sensitizing antibody density on target cell selection by effector monocytes was examined by modifying the sensitization of red cells either by dilution of the antiserum, variation of the number of erythrocytes or both in a cold target competition assay of antibody‐dependent cellular cytotoxicity (ADCC). Human A1 and B erythrocyte target and competitor cells were employed in the presence of hyperimmune anti‐A and anti‐B sera at concentrations above that necessary for saturation of red cells with respect to lytic susceptibility. When the number of red cells was kept constant and the dilution of antisera was varied a linear relationship between the competitive capacity of erythrocytes and the concentration of sensitizing antiserum was observed. When the number of target (competitor) cells and the concentration of antisera were varied simultaneously it was apparent that the competitive capacity was dependent upon the relative densities of the sensitizing antibodies. When competition was tested in the presence of suboptimal concentrations of complement, rather than effector cells it was observed that, in common with ADCC, the effectiveness of cold competition was dependent upon the concentration of sensitizing antibody.
Effects of 3‐Azidothymidine on Platelet Counts, Indium‐111‐Labelled Platelet Kinetics, and Antiplatelet Antibodies
Authors: Panzer Simon , Stain Christoph , Benda Helmut , Mannhalter Christine
Published: March 2009

Abstract. Treatment of the acquired immunodeficiency syndrome with 3‐azidothymidine (zidovudine; AZT) can induce severe neutropenia and anemia, while platelet counts increase. In order to understand the mechanism by which this favorable effect on platelets is induced, we prospectively studied platelet kinetics and platelet serology in 8 hemophiliacs receiving the drug. All patients underwent a second investigation after 3 months of treatment. Four patients were thrombocytopenic before treatment. Platelet counts increased significantly already after 1 week of treatment (p = 0.03), when only 3 patients remained thrombocytopenic. In these latter patients a further increase of platelet counts was noticed during the following 3 months. Platelet life‐span was shortened in all patients at the initial investigation and a significant prolongation was measurable at the second evaluation (p = 0.015). Platelet‐associated immunoglobulin G was increased in 3 patients at the first investigation and in 4 patients at the follow‐up. Platelet‐associated complement (PAC3d) was elevated in all subjects at the first determination. It decreased in 6, but increased in 2 patients thereafter; thus these changes did not become significant (p = 0.078). Immunofluorescence studies revealed antiplatelet antibodies in 7 patients' sera before treatment, and in 5 sera during drug therapy. At both investigations, the antibodies bound to the platelet glycoprotein IIIa as was demonstrated by immunoprecipitation of radiolabeled platelet proteins. We conclude, that AZT treatment improves platelet counts in HIV‐infected hemophiliacs primarily by a prolongation of platelet survival without having a significant influence on antiplatelet antibody binding.
HLA Class I (Bg) Antigens on Red Cells of SLE Patients: A Serological Study with Polyclonal and Monoclonal Antibodies
Authors: Giles Carolyn M. , Darke Christopher , Rowe Graham P. , Botto Marina
Published: March 2009

Abstract. The enhanced HLA class I (Bg) on red blood cells (RBC) of many patients with systemic lupus erythematosus has allowed a significant correlation to be made between their HLA‐B types and haemagglutination reactivity with lymphocytotoxic anti‐HLA‐B sera stimulated by pregnancy alone. Therefore the class I expression on these RBC relates to classical, rather than non‐classical, class I gene products. Studies of class I expression on RBC by means of monoclonal antibodies (MAb) to epitopes on the heavy polypeptide chain and β2‐microglobulin (β2m) have suggested that the complete extracellular structure is present. The specific effect of chloroquine in ‘stripping HLA’ from RBC had been assumed to support the concept that HLA class I was adsorbed from plasma. However, from our data, we conclude that HLA class I is an intrinsic membrane component. We suggest that the action of chloroquine is to remove β2m alone, which prevents normal class I expression and also results in conformational changes to the class I heavy chain, but that it is not capable of removing the membrane‐bound heavy chain.
Thrombocytopenia Following Routine Blood Transfusion: Micro‐Aggregate Blood Filters Prevent Worsening Thrombocytopenia in Patients with Low Platelet Counts
Authors: Lim S. , Boughton B.J. , Bareford D.
Published: March 2009

Abstract. 11 patients with anaemia and thrombocytopenia due to bone marrow failure each received 2 sets of red‐cell transfusions. They were randomised to receive 1 blood transfusion through a standard 170‐μm giving set filter and another through a 40 μm micro‐aggregate filter. After transfusion, the mean fall in platelet count was 15times 109/1 (41.7%) and 1.4times 109/1 (4.6%), respectively (p<0.01). The results confirm the findings of a previous study which showed that micro‐aggregate filters prevent a post‐transfusional decrease in platelet counts. In addition, this new study shows that this is clinically relevant in patients who are thrombocytopenic at the time of their blood transfusion.
Rate of Interaction of IgG1 and IgG3 Sensitized Red Cells with Monocytes in the Phagocytosis Assay
Authors: Brojer Ewa , Merry A.H. , Zupanska Barbara
Published: March 2009

Abstract. Another aspect of the preferential binding to monocytes of IgG3‐coated erythrocytes over IgG1‐coated erythrocytes has been demonstrated by measuring the rate of interaction in the phagocytosis assay. Erythrocytes sensitized with comparable numbers of IgG1 and IgG3 monoclonal and polyclonal anti‐D antibodies were incubated with a monocyte preparation for up to 2 h. Interaction with the erythrocytes sensitized with monoclonal or polyclonal IgG3 was much more rapid than with erythrocytes sensitized with IgG1. With IgG3‐sensitized erythrocytes maximum interaction was found after 30 min, whereas with IgG1‐sensitized erythrocytes 2 h were required for maximum interaction. These results are consistent with the longer hinge region of IgG3 allowing bridges to be formed more easily between the negatively charged erythrocytes and monocytes.
Immunoglobulin Preparation: Safe from Virus Transmission?
Authors: Uemura Yahiro , Yokoyama Kazumasa , Nishida Masayuki , Suyama Tadakazu
Published: March 2009

Abstract. The incidence of non‐A, non‐B hepatitis associated with the administration of immunoglobulin preparations, especially intravenous preparations, which had been considered to be free from virus transmission, is reported. Research efforts to improve the safety of intravenous immunoglobulin preparations which could be administered in a large volume must be continued. Adding sorbitol under weakly acidic conditions, heat treatment of IgG at 60°C for 10 h is possible. Intravenous immunoglobulin preparation manufactured by polyethylene glycol fractionation followed by the heat treatment is not only intact, but also much closer to the ideal intravenous immunoglobulin preparation in the safety and stability.
Metabolic Status of Platelet Concentrates during Extended Storage: Improvement with Pharmacological Inhibitors and Reduced Surface‐to‐Volume Ratio
Authors: Bode Arthur P. , Miller David T.
Published: March 2009

Abstract. The depletion of plasma nutrients and buffering capacity may present a potential barrier to the long‐term liquid storage of platelet concentrates (PC). We have found that PC prepared with reversible inhibitors of platelet activation added to the citrate anticoagulant and stored at a reduced surface‐to‐volume (S/V) ratio have a much slower rate of lactate build‐up (p<0.01), slower consumption of glucose (p = 0.05), and more stable pH (p<0.01) than controls. By pO2 and pCO2 measurements, PC prepared with inhibitors showed evidence of continued respiration and responsiveness even after storage at 22°C for 15 days. In addition, these PC released only 11% of the total cellular LDH during the storage period as compared to the release of 43–67% of the total LDH in control PC. Maximum benefit of the inhibitors was seen after reduction of the S/V ratio of the storage container, which was made possible by the reduced metabolic demands of platelets stored in the unactivated state. These data suggest that the fall in pH and loss of platelet integrity associated with the platelet storage lesion are correlated with a high metabolic rate which can be controlled by inhibiting the activation of platelets during preparation and storage. The use of these inhibitors and reduced bag surface area may make prolonged liquid storage of platelets feasible.
Fatal Pulmonary Transfusion Reaction to Plasma Containing Donor HLA Antibody1
Authors: Eastlund Ted , McGrath Patricia C. , Britten Anthony , Propp Richard
Published: March 2009

Abstract. A 55‐year‐old woman with common variable immunodeficiency and mild chronic obstructive lung disease received 3 units of plasma as immunoglobulin replacement therapy. During the administration of the final unit, her temperature rose 1°C, with no other observable symptoms. Fifteen minutes later she developed shortness of breath without nausea, vomiting, rash, or pruritus. In 30 min she lost consciousness, was breathless, and cyanotic. Resuscitative efforts failed. Autopsy failed to pinpoint a cause of death. There was no evidence of ABO or Rh incompatibility, bacterial contamination, or hemolysis. There were no neutrophil, platelet or IgA antibodies detectable in the patient or the 3 plasma donors. There were no lymphocytotoxic HLA antibodies in the patient or two of the plasma donors. The third donor had HLA‐B35 lymphocytotoxic antibodies that did not agglutinate or aggregate neutrophils. The patient's HLA type was A2, A3; B35, B40. Lymphocytotoxic crossmatches using lymphocytes of the patient were positive with plasma from the third donor but negative with the other two. An eluate prepared from post‐mortem lung parenchymal tissue was cytotoxic to 7 of 8 panel lymphocytes positive for the HLA‐B35 antigen but not with cells lacking B35. The implicated plasma donor was healthy with a history of 6 pregnancies. This case report illustrates the potential hazard of transfusion of plasma containing HLA antibodies.
Evidence for Genetic Linkage between the KEL and YT Blood Group Loci
Authors: Coghlan Gail , Kaita Hiroko , Belcher Elizabeth , Philipps Sylvia , Lewis Marion
Published: March 2009

Abstract. Peak lods (Ẑ) of 3.48 at an estimated recombination fraction of 0.28 derived from 63 male and 90 female meioses indicate linkage between the KEL and YT blood group loci. Consideration is given to two families; a realistic interpretation of the data increases Ẑ to 4.24 at = 0.26.
Comparison of False‐Positive Reactions in Direct‐Binding Anti‐HIV ELISA Using Cell Lysate or Recombinant Antigens
Authors: Craske J. , Turner A. , Abbott R. , Collier M. , Gunson H.H. , Lee D. , Martlew V. , Howell P. , Love E.
Published: March 2009

Abstract. In a 2‐year study of false‐positive anti‐HIV‐1 tests in blood donors at Manchester and Lancaster Blood Banks, the reactions associated with a HIV‐infected cell lysate antigen were compared with those using recombinant‐antigen‐based tests. In year 1 (cell lysate test) at Manchester BTS 0.21% of 119,178 donations were repeatedly reactive, compared with 0.53% of 119,004 donations in year 2 (recombinant antigen). Reactive sera were tested at Manchester PHL by three different immunoassays. Referred specimens were classified as anti‐HIV positive (95–100% reactive in all the assays), equivocal or negative (negative results in all three immunoassays). Two donors were confirmed to be anti‐HIV positive over the 2‐year period. Most sera were negative by confirmatory immunoassays in years 1 and 2. In year 1, a study of 60 referred sera with sex‐ and age‐matched controls showed high correlation between a reactive anti‐HIV‐1 screening test and indeterminate anti‐HIV‐1 patterns on Western blot showing reactions with HIV gag‐coded proteins. In year 2, less than 10% of referred sera were reactive by Western blot, and there was no correlation between a reactive screening anti‐HIV test, the strength of signal in the test or a reactive Western blot. Follow‐up showed that donors whose sera were reactive in years 1 and 2 by the anti‐HIV‐1 screening test formed almost two different populations. Four donors with equivocal anti‐HIV‐1 confirmatory tests had anti‐HIV ‘envelope’ reactions. The follow‐up of donors with equivocal confirmatory immunoassays on Western blot failed to show an increase in the strength of immunoassay reaction or Western blot reaction, with the possible exception of 1 donor with an anti‐HIV‐p24 reaction by Western blot. Most confirmatory testing can be done using immunoassays. The significance of the Western blot reactions needs further investigation.
Alloimmunization to RhD by Platelet Transfusions in Autologous Bone Marrow Transplant Recipients
Authors: McLeod Bruce C. , Piehl Michael R. , Sassetti Richard J.
Published: March 2009

Abstract. Platelet transfusions from RhD‐positive (D‐positive) donors are often given to RhD‐negative (D‐negative) cancer patients. The low observed rate of alloimmunization has been attributed to disease and therapy‐related immunosuppression. We have studied the occurrence of alloimmunization in 16 D‐negative patients who did not have detectable anti‐D prior to autologous bone marrow transplantation for malignant disease. All received D‐positive platelets, but no other D‐positive blood product. Three patients (19%) developed anti‐D at 13, 24 and 83 days, respectively, after first receiving D‐positive platelets, and after a total dose of 53, 65 and 119 D‐positive platelet unit equivalents, respectively. Two of them also developed anti‐C. The 13 patients in whom anti‐D was not detected were also heavily transfused with D‐positive platelets (mean ± SD = 136 ± 82 platelet unit equivalents). In 6 of them, the last recorded antibody screen was less than 3 months after the first D‐positive platelets, and may not exclude a primary immune response. Thus, despite profound immunosuppression associated with autologous marrow transplantation, alloimmune responses to D‐positive red cells in platelet concentrates can occur in some D‐negative recipients.
Evaluation of Monoclonal Antibodies with Putative Specificity for Human IgG Allotypes
Authors: Nelson P.N. , Fletcher S.M. , de Lange G.G. , van Leeuwen A.M. , Goodall M. , Jefferis R.
Published: March 2009

Abstract. Fourteen monoclonal antibodies (MAbs) of putative specificity for human IgG allotypes and isoallotypes were evaluated for reactivity and specificity in 8 different assay systems. The study showed that the MAbs tested could be classified into 1 of 4 groups: those exhibiting allotypic specificity regardless of the assay system, allotypic specificity dependent on the assay system, isoallotypic specificity, and those showing neither allotypic nor isoallotypic specificity. These observations were presumably dependent on antigen presentation, epitope integrity and/or antibody multispecificity. For the G1m(a), G1m(f), G1m(z), G3m(g) and G3m(u) specificities, MAbs have been produced which can be used for routine typing purposes in defined haemagglutination and enzyme‐linked immunosorbent assay systems. MAbs are also available that show ‘non‐g’ isoallotypic specificity.
Investigation of Lewis Phenotypes in Polynesians: Evidence of a Weak Secretor Phenotype
Authors: Henry S. M. , Benny A. G. , Woodfield D. G.
Published: March 2009

Abstract. The salivary ABH and Lewis antigens of Polynesians were measured using a standardised red cell agglutination microplate assay and compared with the red cell defined Lewis phenotypes. Salivary ABH substances were detected in almost all saliva samples tested, with low levels (partial secretion) of ABH substances in the saliva from Le(a+b‐) and Le(a+b+) individuals. Salivary Leb substance was detected in all Le(a‐b+) and Le(a+b+) samples and in almost all Le(a+b‐) samples. It is evident from the results obtained that Polynesian red cell phenotypes cannot be used to predict the presence or absence of salivary substances.

If the presence of a coding secretor gene is presumed responsible for salivary ABH antigens and salivary Leb antigen expression, then the incidence of a coding secretor gene in Polynesians is 98%. These results indicate that the recessive non‐secretor gene is absent or rare in a Polynesian derived gene pool. Two variants of secretor individuals are found among Polynesians, secretors with expression of normal amounts of the product of the secretor gene, similar to Caucasians, and partial secretors with weak expression of the secretor gene products.
Human Monoclonal Anti‐D with Reactivity against Category DVI Cells Used in Blood Grouping and Determination of the Incidence of the Category DVI Phenotype in the DU Population
Authors: Leader K. A. , Kumpel B. M. , Poole G. D. , Kirkwood J. T. , Merry A. H. , Bradley B. A.
Published: March 2009

Abstract. B‐lymphoblastoid cell lines transformed by Epstein‐Barr virus were produced from cells obtained from a hyperimmunised donor with serum anti‐D activity against category DVI red cells and enriched for this activity by rosetting with category DVI red cells. Three clones produced IgGl anti‐D and had stable cell growth and continuous secretion of antibody in prolonged culture. The monoclonal antibodies reacted with category DVI red cells, when assessed manually and in an automated blood grouping system, and are useful blood grouping reagents for the detection of the category DVI phenotype. Using a radiometric technique, the number of antibody molecules bound to category DVI red cells from 5 individuals was estimated to range from 2,800 to 11,200 per cell. Five percent of blood donors classed as Du in the south western region were found to have the category DVI phenotype.
Relationship of Inb Antigen to Other Antigens on In(Lu)‐related p801
Authors: Telen Marilyn J. , Ferguson David J.
Published: March 2009

Abstract. The Ina and Inb antigens have recently been added to the list of diverse erythrocyte blood group antigens whose expression is down‐regulated by the In(Lu) gene. Evidence has been provided that these antigens constitute polymorphisms of an erythrocyte glycoprotein previously defined by its reactivity with murine monoclonal antibodies, such as A3D8, and designated In(Lu)‐related p80. This study has now confirmed the assignment of the Inb antigen to p80 and has further studied the relationship of the Inb epitope to other p80 epitopes recognized by both monoclonal antibodies as well as polyclonal antiserum produced against purified erythrocyte p80.