bluuebunny/crossref_metadata_2025 huggingface dataset:
Platelet Transfusion Reaction Associated with Interdonor HLA Incompatibility1
Authors: Eastlund D. Ted , McGrath Patricia C. , Burkart Peter
Published: March 2009

Abstract. An HLA‐compatible platelet transfusion was followed by chills, fever, and severe respiratory distress in a multitransfused patient with chronic lymphocytic leukemia. During the previous 7 days the patient had received blood products without incident, including 8 units of red blood cells (RBC), 24 units of pooled random donor platelet concentrates, and five HLA‐compatible platelet pheresis products. The patient had no demonstrable RBC, HLA lymphocytotoxic, platelet or granulocyte antibodies. The platelet donor, a multiparous female, had no granulocyte or RBC antibodies but had lymphocytotoxic antibodies against HLA‐A2 CREG (cross‐reacting group A2, A28, A23, A24) which reacted not with lymphocytes of the patient but with lymphocytes of the donor whose RBC were transfused 24 h prior to the platelet transfusion reaction and whose HLA type is A23, A24; B44, B57. No RBC donors had HLA lymphocytotoxic, granulocyte, or platelet antibodies against the platelet donor. The patient received three subsequent platelet transfusions from the same donor after removal of the antibody‐laden plasma with no adverse reaction. These data suggest an interdonor reaction caused by the presence of cells from the RBC donor received by the patient 24 h prior to the transfusion of donor lymphocytotoxic antibody to HLA‐A2 CREG antigens.
Early Immunization against Platelet Glycoprotein Ilia in a Newborn Glanzmann Type I Patient
Authors: Bierling Philippe , Fromont Patricia , Elbez Annie , Duedari Najib , Kieffer Nelly
Published: March 2009

Abstract. Alloimmunization against platelet glycoprotein lib and/or IIIa is a complication rarely observed during the evolution of type I Glanzmann's thrombasthenic patients [1,2]. The occurrence of such alloantibodies is usually due to repeated blood transfusions [2, 3] and greatly complicates the treatment of these patients since they prevent effective platelet transfusion and might, theoretically, cause posttransfusion purpura. We describe the case of a newborn thrombasthenic patient who developed an IgG platelet allo‐antibody 1 month after birth. The diagnosis of Glanzmann's thrombasthenia was complicated by the rare platelet phenotype (PLAt‐negative PLA2‐positive) of the healthy mother, which was probably heterozygous for the abnormal thrombasthenic gene. Immunofluorescence and immunoblotting techniques demonstrated that the patient antibody was principally directed against the platelet glycoprotein IIIa. Surprisingly, this patient had only received four blood transfusions (fresh frozen plasma on days 1 and 2, and standard red blood cell concentrates on days 5 and 6) before the discovery of the antibody, suggesting prior in utero sensitization. This study emphasizes the need for early diagnosis of the disease. Thrombasthenic patients should be transfused with deleukocyted platelet‐free blood products.
Platelet Anti‐BAKb Antibody Associated with Neonatal Alloimmune Thrombocytopenia
Authors: McGrath K. , Minchinton R. , Cunningham I. , Ayberk H.
Published: March 2009

Abstract. A platelet alloantibody of IgG class but unknown specificity was detected in the serum from the mother of an infant with neonatal alloimmune thrombocytopenia. The antibody reacted by indirect immunofluorescence with 33/65 random donors, 10/11 Baka‐negative donors and 23/54 Baka‐positive donors. The mother's platelet phenotype was PLA1, Koa, Kob, Baka and the father's was PLA1, Koa, Kob, Baka. Immunoblotting and immunoprecipitation of maternal serum with paternal platelets produced a band at molecular weight 140 kilodaltons identical to the band obtained with Baka antiserum. Family studies confirmed the allelic distribution of Baka and the unknown antigen. The platelet‐specific antibody in this patient meets the criteria for an antibody to the new platelet antigen, Bakb.
Characterization of a Neuraminidase from Corynebacterium Aquaticum Responsible for Th Polyagglutination
Authors: Sondag‐Thull D. , Levene N.A. , Levene C. , Manny N. , Liew Yew Wah , Bird G. W G. , Schechter Y , François‐Gérard Ch. , Huet M. , Blanchard D.
Published: March 2009

Abstract. Th polyagglutinability is characterized by the agglutination of the red blood cells (RBC) by Arachis hypogaea, Medicago disciformis, Vicia cretica but, in contrast to the T phenomenon, not by Glycine max (Glycine soja). Because Th transformation of RBC has been obtained in vitro, the mechanism of Th polyagglutinability expression has been studied and reproduced experimentally. An enzyme with neuraminidase specificity has been isolated from the culture supernatant of Corynebacterium aquaticum, and further characterized (MW = 55,600 kDa, pH = 5.5, Km = 0.138 μM, Kcat = 0.22 μg). Reversely, Th transformation of RBC could be obtained by using other neuraminidases but in very mild conditions of hydrolysis. From our results, it can be concluded that by the release of less than 20 μg of sialic acid per 1010 RBC, Th reactivity can be induced whereas hydrolysis of greater amounts of sialic acid (>20 μg/1010 RBC) give the classical T polyagglutinability.
Severely Heated Therapeutic Factor VIII Concentrate of High Specific Activity
Authors: Winkelmana L. , Owena N. E. , Evans D. R. , Evans H. , Haddon M. E. , Smitha J. K. , Prince P. J. , Williams J. D. , Lane R. S.
Published: March 2009

Abstract. A new method for the manufacture of a heated factor VIII concentrate of high specific activity (2–6 IU factor VIII:C/mg protein) has been developed. Addition of heparin to cryoprecipitate extract at acid pH precipitated fibrinogen and fibronectin. Factor VIII was then recovered from the supernatant by precipitation with glycine and sodium chloride. After re‐solution and desalting on Sephadex G‐25, the concentrate was sterile‐filtered and lyophilised. The dried product was stable to heating in the final container at 80°C for 72 h. Data from 25 consecutive batches of concentrate prepared from 1,200‐1,500 kg plasma pools are presented. The mean final yield of heated product was 190 IU factor VIII:C/kg plasma. The concentrate has been found to be safe and effective in clinical use.
Storage of Platelets in Earth Orbit. Effect of Gravity and the Formulation of Plastic Storage Containers on Platelet IgG and C31
Authors: Szymanski Irma O. , Odgren Paul R. , Teno Richard A. , Jacobson May S. , Surgenor Douglas MacN.
Published: March 2009

Abstract. We investigated the effects of gravitational force and three different plastic formulations of the storage bags on the quality of stored platelets by measuring the changes in platelet‐associated immunoglobulin G (IgG) and two antigenic markers of the third component of human complement (C3), C3d and C3c. Pooled platelets were stored in parallel at microgravity (MG) and on the ground (1g) using three different plastic polymers: (1) polyvinylchloride (PVC) plasticized with di‐2‐ethylhexyl phthalate (DEHP); (2) PVC plasticized with trioctyl trimellitate (TOTM), and (3) unplasticized polyolefin (PO). The IgG and C3 were quantified by an automated antiglobulin consumption test in freeze/thaw disrupted platelets (total IgG or C3). The baseline values for platelet associated IgG and C3, measured after 2.5 days of storage at 1g just prior to the launch, fell within the normal range. Following an additional 6.5 day storage, platelets stored at MG had accumulated significantly less C3d than those stored at 1g, suggesting that MG storage was beneficial. Specific plastic formulations also exerted a significant effect on the accumulation of these immunoproteins, the effect being particularly pronounced at MG. The smallest increases of IgG and C3 were seen in platelets stored in TOTM and the largest in those stored in DEHP. It is possible that further studies at MG would permit a clear characterization of the effects of other independent storage variables.
False Negative Results in a Competitive Immunoassay for Anti‐HIV‐Antibodies Due to the Presence of Rheumatoid Factors
Authors: Janvier D. , Reviron M. , Reviron J.
Published: March 2009

Abstract. One serum from a blood donor, which had both HIV‐antibodies and rheumatoid factor, gave positive results with EIA anti‐HIV screening tests and Western blot, but negative results with the Abbott's ENVACOR assay. Moreover, this serum and 5 out of 6 sera containing only rheumatoid factors were able to interfere with ENV and CORE reactions. Rheumatoid factors bind to the beads used in ENVACOR test and are also able to react with the conjugate. Most of the rheumatoid factors bind to the ENV conjugate, though with a wide range of activity. Obvious binding of rheumatoid factors to the CORE conjugate could not be demonstrated. Binding of rheumatoid factors both to the bead and to the conjugate leads to high optical density values and therefore results in the negative range according to the normal interpretation of the ENVACOR competition procedure.
Centrifugation of Very Freshly Donated Blood May Yield Platelets Unstable to Storage in the New Generation of Containers
Authors: Solberg Christel , Hansen John‐Bjarne , Little Clive
Published: March 2009

Abstract. The stability of platelets stored in the second generation of container has been examined further. Platelet concentrates prepared in CPD anticoagulant were found to deteriorate fairly rapidly during storage if prepared from very freshly donated blood, whereas a more than 4‐hour delay between donation and platelet isolation yielded platelet concentrates much more stable to storage. Delays of more than 2 h (relative to the time of donation) in the isolation of platelets led to decreases in the aggregation response to low or moderate doses of collagen. Platelets from blood collected into CPD adenine showed decreased aggregation responses relative to equivalent cells from blood collected into adenine‐free anticoagulant. It seems that a decreased aggregation response is associated with improved storage properties.
Screening for AIDS in Western India: Transfusion Aspects
Authors: Rodrigues Jeanette J. , Israel Zimra R. , Athalye Neelima , Pavri Khorshed M.
Published: March 2009

Abstract. Screening of blood donations for anti‐HIV in western India has shown a rising trend of antibody‐positive donors. There was a statistically significant difference between the proportion of positive reactors before 1986 and after 1986 (p<0.05).
Posttransfusion Purpura Associated with an Anti‐Bakb
Authors: Kiefel V. , Santoso S. , Glöckner W.M. , Katzmann B. , Mayr W.R. , Mueller‐Eckhardt C.
Published: March 2009

Abstract. We describe a 46‐year‐old white woman with typical clinical features of posttransfusion purpura (PTP) whose serum held a platelet‐specific alloantibody reactive with an antigen antithetical to Baka, i.e. anti‐Bakb. The specificity of the antibody was confirmed by family analysis, a population study (expected versus observed gene frequency: 0.3651 versus 0.3984; n=105) and localization of the antigen on glycoprotein IIb in radioimmunoprecipitation. Typing of family members and blood donors for platelet antigens disclosed that the patient had been preimmunized by two blood transfusions in 1981, while fetomaternal incompatibility for Bakb was ruled out (her three children and their father were Bakb negative). Treatment of PTP with corticosteroids and platelet transfusions was ineffective, but infusion of high‐dose intravenous IgG resulted in a rapid increase in the platelet count.
A Cold Agglutinin: Om
Authors: Kajii Eiji , Ikemoto Shigenori
Published: March 2009

Abstract. An IgMk cold agglutinin reacted with cord blood as well as with adult red blood cells (RBC). The agglutinin reacted strongly with papainized and neuraminidase (RDE)‐treated RBC. It resembled anti‐Me cold agglutinin in these serological findings. However, the activity of this agglutinin against RBC was markedly decreased in the presence of human milk. The data suggest that the agglutinin is directed against a new glycolipid antigen common to human RBC, termed Om antigen.
Immunoglobulin G Subclass Distribution in Three Human Intravenous Immunoglobulin Preparations
Authors: Burckhardt Johann J. , Gardi Andreas , Oxelius Vivi‐Anne , Preud'homme Jean‐Louis , Scherz Rudolf , Skvaril Frantisek , Heiniger Hans‐Jörg
Published: March 2009

Abstract. In immunodeficiency patients the lack of immunoglobulins (Ig) can be total or partial with a specific IgG subclass imbalance masked by normal values for total IgG. In the latter case therapy with intravenous IgG preparations (IVIG) is generally beneficial, provided the IVIG preparations used originate from large pools of normal blood donors and exhibit a normal IgG subclass distribution. We have analyzed the subclass distribution of three IVIG products: Sandoglobulin® (SAGL), Gamimune®N (GI), Gammagard® (GG), 6–10 lots each, in four different laboratories. The competitive enzyme immunoassays and radial immunodiffusion methods used different monoclonal and polyclonal antibodies specific for IgG1, IgG2, IgG3, and IgG4, respectively. Despite minor interlaboratory differences, the results show that the slightly lower IgG1 content of SAGL versus GI and GG was quantitatively compensated by a higher proportion of IgG2, that no differences existed in IgG3 levels, but that one preparation (SAGL) contained 2–3% of IgG4 compared to 0.5–1.5% in GI and below 0.5% in GG. This difference was significant, the two latter preparations being at or below the lower limit of what are considered to be normal values found in human adults. Such differences may have important clinical consequences.
Evaluation of 99mTechnetium/51Chromium Post‐Transfusion Recovery of Red Cells Stored in Saline, Adenine, Glucose, Mannitol for 42 Days1
Authors: Heaton W.A.L. , Keegan T. , Holme S. , Momoda G.
Published: March 2009

Abstract. An in vivo and in vitro study of 42‐day saline, adenine, glucose, mannitol (SAGM) red cells (Terumo Corporation, Elkton, Md.) was conducted using 20 volunteers to document in vivo efficacy and analyze the validity of the 99mTc/51Cr technique. In vivo autologous post‐transfusion recovery was measured by a double‐label procedure involving the use of 99mTc‐labeled freshly drawn red cells to quantify recipient blood volume and 51Cr to document both posttransfusion 24‐hour recovery (PTR) and red cell survival. As an internal control, 5 of the 20 donors studied had red cell volumes estimated by the 125I‐albumin (RISA) plasma volume technique on a separate occasion. For comparison, PTR was also calculated according to the single‐label 51Cr protocol in which recipient blood volume was estimated by extrapolation from circulating 51Cr‐labeled red cell activity 5–15 min after infusion. After 42 days of storage, PTR averaged 78±6% with the double‐label 99mTc/51Cr technique and 81±5% with the single‐label (51Cr alone), confirming a small difference between the two techniques. Correlation between the two techniques was high, r = 0.81, though the average 3% difference between them was significant (p<0.05). Red cell volumes of the 5 donors measured by both the 99mTc‐red cell method and the 125I‐albumin method exhibited excellent correlation, r = 0.96, and averaged 1,961 ±420 ml and 2,048±381 ml, respectively. Standard in vitro parameters of SAGM red cell concentrates were well preserved with hemolysis of 0.5±0.4% and red cell ATP 69±13% of the initial values after 42 days of storage. The double‐label, 99mTc/51Cr, post‐transfusion recoveries exhibited a slightly better correlation with post‐storage ATP when compared to single‐label, 51Cr, recoveries: r = 0.705 versus r = 0.644, respectively. The results confirmed acceptable 42‐day storage parameters for modified SAGM red cells.
An Epidemiological and Clinical Study of Transaminase Levels and Hepatitis B Antibodies in 1,100 Blood Donors
Authors: Driss F. , Boboc B. , Zarski J.P. , Cals M.J. , Pol S. , Eme D. , Ekindjian O.G. , Courouce A.M. , Brechot C. , Berthelot P. , Nalpas B.
Published: March 2009

Abstract. In order to provide epidemiological and clinical information on surrogate testing of blood donations, the respective prevalences of serum hepatitis B virus (HBV) markers and elevated transaminase levels were studied in 1,100 blood donors according to their geographic origin and socioeconomic level. The frequency of serum HBV markers varied as a function of HBV endemicity in the country of origin; however, it was inversely correlated (p>0.05) to the socioeconomic level of the donors, even in those originating from countries of low HBV endemicity. There was no association between serum HBV markers and the increased transaminase level which was observed in 48 (4.3%) donors. Twenty‐five of these accepted further clinical evaluation. A diagnosis appeared probable in 12 of the 25: alcohol in 5; drugs in 6; non‐A, non‐B hepatitis in 1. Seven of the remaining 13 subjects were more than 25% above ideal body weight. Transaminase activities determined at the time of clinical assessment were normal in 14. In addition, serum HBV DNA was found in 5 of 247 donors, even in the absence of any usual HBV marker and/or hypertransaminasemia. This could account for the few cases of B and B‐like posttransfusion hepatitis which are known to still occur despite careful HBsAg screening of blood donors.
A Private Red Cell Antigen, Jones, Causing Haemolytic Disease of the Newborn
Authors: Reid M. , Fischer M. L. , Green C. , Rowe G. P. , Pfister M. , Poole J. , Lewis M.
Published: March 2009

Abstract. Two previously unpublished low‐incidence antigens, Jones and Hol., are identical. The antigen is a dominant, autosomally inherited character that segregates independently from the loci for the ABO, MNS, Duffy and Yt blood group systems and is different from previously published infrequent antigens. The antigen is apparently unaffected by enzyme treatment and is well developed on red cells of neonates. The antibody reacts best by indirect antiglobulin testing, is IgG and has caused haemolytic disease of the newborn. This private blood group antigen, named Jones, has been assigned the ISBT number 700047.
Coexpression of Sialosyl‐Tn (NeuAcα2→6GalNAcαl→0‐Ser/Thr) and Tn (GalNAcα1→O‐Ser/Thr) Blood Group Antigens on Tn Erythrocytes1
Authors: Kjeldsen T. , Hakomori S. , Springer G. F. , Desai P. , Harris T. , Clausen H.
Published: March 2009

Abstract. Polyagglutinable erythrocytes expressing the rare human blood group phenotype Tn were tested for expression of both Tn (GalNAcα1→O‐Ser/Thr) and sialosyl‐Tn (NeuAcα2→6GalNAcα1→O‐Ser/Thr) antigens by agglutination with a panel of well‐characterized monoclonal antibodies specifically directed to Tn and sialosyl‐Tn antigens, respectively. Tn erythrocytes from 4 patients were strongly agglutinated by both sialosyl‐Tn and Tn‐specific monoclonal antibodies, indicating coexpression of sialosyl‐Tn and Tn antigens on the cell membrane surface of Tn polyagglutinable erythrocytes. Human polyclonal anti‐Tn antisera were found to express weak and variable anti‐sialosyl‐Tn antibodies in conjunction with the anti‐Tn antibodies. The Tn phenotype thus involves not only the classically known Tn antigens but also sialosyl‐Tn antigens. However, the human anti‐Tn antibodies may be directed mainly toward the Tn antigen (GalNAcα1→O‐Ser/Thr).
Autoanti‐Ge Associated with Severe Autoimmune Hemolytic Anemia
Authors: Shulman Ira A. , Vengelen‐Tyler Virginia , Thompson Joseph C. , Nelson Janice M. , Chen David C. T.
Published: March 2009

Severe autoimmune hemolytic anemia due to anti‐Ge is described. The patient's red cells had a positive direct antiglobulin test, and they typed as Ge+ using saline reactive reagents. Anti‐Ge was eluted from her RBCs, and her serum had an IgG and IgM anti‐Ge2,3.
Storage of ADSOL‐Preserved Red Cells at 2.5 and 5.5°C: Comparable Retention of in vitro Properties1
Authors: Moroff Gary , Holme Stein , Keegan Thais , Heaton Andrew
Published: March 2009

Abstract. Previous studies with CPDA‐1 and Nutricel preserving solutions indicated that red cell properties were affected by small differences in storage temperature. The influence of a 3°C differential on the preservation of the in vitro properties of ADSOL‐preserved (AS‐1) red blood cells was investigated in a paired study. AS‐1 red blood cells were stored at 2.5 and 5.5°C for 42 days. Extent of hemolysis, glucose consumption and pH levels were comparable at the two storage temperatures. Lactate levels were slightly, but significantly higher at 5.5°C. ATP levels were slightly but significantly higher at 2.5°C, only during the later part of the storage period. 2.3‐DPG levels were slightly better retained at 2.5°C after 7 days of storage. Holding units of whole blood for either 1 or 8h at ambient temperature after phlebotomy prior to processing did not influence the types of temperature‐dependent changes. The differences as a function of storage temperature were small and appear to be of no practical importance in connection with the storage of AS‐1 red blood cells.
Anti‐Rh(D) IgG Subclasses and Severity of Rh Hemolytic Disease of the Newborn
Authors: Pollock Janet M. , Bowman John M.
Published: March 2009

Abstract. The influence of anti‐Rh(D) IgG subclasses on the severity of hemolytic disease of the newborn was examined in 98 pregnancies. Disease was severe in 19 of 32 cases in which only IgG1 was detected, 40 of 63 cases in which IgG1 and IgG3 were found and 0 of 3 cases in which only IgG3 was detected. When both IgG1 and IgG3 anti‐D were present, hemolytic disease was more severe than when IgG1 alone was detected. The earliest onset of hydrops was at 20 weeks gestation in the IgG1+IgG3 anti‐Rh group, and at 27 weeks gestation in the IgG1 group.
Management of Severe Hemolytic Disease with Ultrasound‐Guided Intravascular Fetal Transfusions
Authors: Kanhai H.H.H. , Gravenhorst J. Bennebroek , van Kamp I.L. , Meerman R.H. , Brand A. , Dohmen‐Feld M.W.A. , Ruys J.H.
Published: March 2009

Abstract. Between January 1987 and March 1989, 22 fetuses with severe hemolytic disease were treated with 64 ultrasound‐guided intrauterine intravascular transfusions. Eighteen infants survived and are doing well. In 12 fetuses, hydropic changes were present at the first transfusion; 9 of these survived. In 8 of the 10 fetuses with a gestational age < 26 weeks, intrauterine treatment was successful.
An Improved Assay for Human Tetanus Anti‐Toxin and Its Use in the Accession of Human Plasma for the Production of High‐Titre Tetanus Immunoglobulin
Authors: Kenrick K. G. , Wallace R. C. , Ismay S. L.
Published: March 2009

Abstract. A simplified passive haemagglutination (PHA) screening test, an improved quantitative PHA assay, and a stable test cell preparation are described, as well as a comprehensive testing strategy which have been used in concert at this Service over the past 10 years for the successful accession of high‐titre tetanus anti‐toxin (TAT) plasma for fractionation into human tetanus immunoglobulin (HTIG). The sequential deployment of the screening and quantitative assays, has permitted large numbers of donors to be screened quickly and economically, and has helped establish a significant core of regular donors with high TAT levels. The assays have proven to be highly sensitive and specific and relatively simple to perform, while the coated cells are inexpensive and easily prepared. Approximately 20% of donors screened from the Sydney metropolitan area had TAT levels of 3 IU/ml or greater.
Studies in Red Blood Cell Preservation 1. Effect of the Other Formed Elements
Authors: Greenwalt T. J. , Sostok C. Zehner , Dumaswala U. J.
Published: March 2009

Abstract. The purpose of this study was to determine if the removal of most of the leukocytes and platelets would affect the in vitro characteristics of stored red blood cells (RBC). Fresh RBC concentrates prepared by removing platelet‐rich plasma and filtration through Imugard IG 500 filters were compared with unmanipulated units after storage for up to 56 days. The filtered units were significantly better after 56 days storage for supernatant K+ (p = 0.001), hemolysis (p = 0.05), total vesicle membrane protein shed (p = 0.03), and RBC morphology score (p = 0.04). These differences occurred even though ATP levels were well maintained in both groups. The measurements that did not differ significantly were pH, hematocrit, ATP, 2,3‐DPG, glucose and supernatant Na+. It is suggested that enzymes, leukotrienes, catecholamines and eicosanoids released by degenerating leukocytes and platelets may be inimical to RBC. Some may act as agonists on a‐adrenergic and cholinergic muscarinic receptors present on RBC membranes.
Immunoblotting of Human Red Cell Membranes: Detection of Glycophorin B with Anti‐S and Anti‐s Antibodies
Authors: Khalid G. , Green C.A.
Published: March 2009

Abstract. Human anti‐S and anti‐s eluates bound to glycophorin B on immunoblots from membranes of S+ and s+ red cells, respectively. Eluates of human anti‐S were more efficiently prepared from sensitized trypsin‐treated cells than from sensitized untreated cells. The results of immunoblotting membranes from enzyme‐treated cells confirmed the serological findings: S and s antigens were not affected by treatment with trypsin or sialidase but were destroyed or much depressed by treatment with papain, pronase or α‐chymotrypsin. Immunoblotting with anti‐S or anti‐s of membranes from cells with unusual MNS phenotypes confirms the involvement of glycophorin B in hybrid glycophorins; the existence of such hybrid glycophorins was deduced previously from serological work or immunoblotting with monoclonal antibodies. The presence of s‐active glycophorin B in glycophorin (B‐A)Dantu, in glycophorin BMiIII and in glycophorin (A‐B)MIV was confirmed. The bands observed when Mit+ membranes were immunoblotted with anti‐S supports the suggestion from serological work that the Mit antigen is associated with an altered S antigen.
A Highly Purified Factor VIII:c Concentrate Prepared from Cryoprecipitate by Ion‐Exchange Chromatography
Authors: Burnouf T. , Burnouf‐Radosevich M. , Huart J. J. , Goudemand M.
Published: March 2009

Abstract. A new ion‐exchange chromatographic procedure has been developed to produce a highly purified factor VIII (FVIII) concentrate from plasma cryoprecipitate. Solubilized cryoprecipitate, after adsorption on aluminium hydroxide and cold precipitation, was treated with 0.3% tri(n‐butyl)phosphate and 1% Tween 80 at 25°c for at least 8 h to inactivate lipid‐enveloped viruses. The fraction was then loaded onto a column packed with DEAE‐Fractogel TSK 650 M and chromatographed. Most proteins and TnBP‐Tween 80 flowed through the gel unretarded. FVIII:c, which bound to the gel, was eluted by increasing the ionic strength, then was directly filter‐sterilized without ultrafiltration or addition of a protein stabilizer. Chromatographic recovery of FVIII:c was 80–90%. After freeze‐drying, FVIII:c was at a concentration of 42.5 ± 9.5 IU/ml and had a specific activity of 175.4 ± 37.8 IU/mg (n = 40), corresponding to a purification factor of over 12,000 from plasma. The typical yield of the freeze‐dried FVIII:c from cryoprecipitate was 55–65%. FVIII:c was stable for over 24 h at room temperature in the liquid state. The mean content of fibrinogen and immunoglobulin G was only 65 and 100mg/l, respectively, corresponding to 1.4 and 2.3 mg/1,000 IU FVIII:c. This concentrate, which is much purer than traditional FVIII concentrates, has been found to be well tolerated and effective in clinical treatment of hemophilia A patients.
Evidence That the Human Blood Group Antigens Gya and Hy Are Carried on a Novel Glycosylphosphatidylinositol‐Linked Erythrocyte Membrane Glycoprotein
Authors: Spring F. A. , Reid M.E.
Published: March 2009

Abstract. Immunoblotting under non‐reducing conditions with purified human anti‐Gya and anti‐Hy locates both antigens to an erythrocyte membrane glycoprotein of apparent Mr 46,750–57,500. The antigens are destroyed on intact red cells by the enzymes pronase, trypsin and chymotrypsin, and by treatment with reducing agents. Immunoblotting with anti‐Gya and anti‐Hy to membranes prepared from red cells pre‐treated with an Endo F preparation caused a mean reduction in apparent Mr of the glycoprotein by 11kDa at the leading and trailing edges, when compared with control membranes. These results suggest that the glycoprotein has one or more complex N‐glycans that are not completely sensitive to Endo F digestion on intact cells. The majority of Gya/Hy‐active molecules are not tightly associated with the red cell membrane skeleton. A gross reduction in reactivity with anti‐Gya and anti‐Hy by immunoblotting was observed in red cell membranes from patients with paroxysmal nocturnal haemoglobinuria, suggesting a possible membrane linkage via glycosylphosphatidylinositol for the glycoprotein that carries the Gya and Hy antigens. Immunoprecipitation of the glycoprotein by anti‐Gya showed that the protein migrates faster under reducing conditions (Mr 45,000–54,000). A putative dimer was also evident in the precipitates. The glycoprotein was demonstrated to be distinct from lymphocytefunction‐associated antigen‐3 (CD58), the LWab‐active glycoprotein, the Fya‐active glycoprotein, the Oka‐active glycoprotein and the BRIC 125 glycoprotein (CD47).
Current Status of Fibronectin in Transfusion Medicine: Focus on Clinical Studies
Authors: Powell Frank Stanton , Doran Jan Eva
Published: March 2009

Abstract. It has long been hypothesized that fibronectin (Fn) is essential to the function of the reticuloendothelial system (RES) and that the reversal of Fn deficiency in critically ill patients would result in a clinical benefit to these patients. Fn administration to deficient patients was postulated to improve the function of the RES, decrease the incidence of organ failure, sepsis and ultimately mortality. Over the past 15 years, several clinical studies have been performed to test these hypotheses. The initial anecdotal studies using cryoprecipitate (a plasma fraction enriched in Fn) revealed promising results but were neither controlled nor blinded. Further controlled studies were published utilizing both cryoprecipitate and purified Fn. Unfortunately, the great majority of authors found no beneficial effects of Fn administration in critically ill patients, in relation to incidence of organ failure, sepsis, or mortality. These results do not support the use of Fn in this setting.

Fn utilization in wound healing has shown promising results in case reports. Although its role in wound healing is not yet fully delineated, initial reports with corneal wounds show a beneficial influence of Fn administration. Further studies are needed to determine the exact function(s) of Fn in a healing wound. Efficacy must still be shown in controlled clinical trials; dosing and administration regimens need to be elucidated.
Leukocyte Filtration Removes Infectious Particulate Debris but Not Free Virus Derived from Experimentally Lysed HIV‐infected Cells
Authors: Rawal Bhupat , Yen T. S. Benedict , Vyas Girish N. , Busch Michael
Published: March 2009

Abstract. We used in vitro model of storage‐induced leukocyte degradation in blood to experimentally characterize the infectivity of the debris of lymphocytes harboring human immunodeficiency virus (HIV‐1). The leukocyte filtration process removed both intact HIV‐infected H9 cells and the particulate debris, but failed to remove the cell‐free HIV released from lysed cells. These data suggest that the leukocyte filtration of donor blood soon after collection would optimally reduce the particulate infectious burden of blood for transfusions.
A New Qualitative Variant of the RhE Antigen Revealed by Heterogeneity among Anti‐E Sera
Authors: Lubenko A. , Burslem S.J. , Fairclough L. M. , Pope J. , Boyd D. , Bacon J.
Published: March 2009

Abstract. We have recently detected a new qualitative variant of the RhE antigen characterised by negative reactions with a minority of anti‐E sera, and an inability of the relevant cells to completely absorb most, if not all, anti‐E samples that we tested. In tube tests, using a two‐stage enzyme technique, cells from the proposita (V.R.) were completely agglutinated by 8 out of 12 undiluted polyclonal sera, weakly agglutinated (2+ and 3+ reactions) by 2 sera, but not agglutinated by the 2 remaining sera. Cross‐absorption tests using 1 non‐reacting, 1 weakly reacting and 4 strongly reacting polyclonal anti‐E sera showed that V.R.‘s cells only slightly reduced their anti‐E titres versus r“r cells, whilst completely removing their reactivity against her own cells. Absorptions with R2R2 cells abolished activity against both cell types. The cells of V.R. failed to absorb or elute anti‐Ew, and had normal expression of c. Three monoclonal human anti‐E reagents were tested by fluorescence flow cytometry; one reagent failed to react with the cells of V. R., a second bound as strongly with V.R.'s cells as with R1R2 controls while the third bound much more weakly with V.R. than with any other E+ sample. The variant of RhE in the V.R. family is therefore distinct from Ew, and lacks an epitope recognised by a major constituent of most anti‐E sera. This epitope is unlikely to be ET, since all of 91 samples from E‐positive Australian aborigines (many of which are expected to lack ET) were agglutinated by a serum that failed to react with V.R.‘s cells. No other V.R.‐like E variants were found on prospective screening of 1,006 E+e+ North London donors with 1 strongly and 1 non‐reacting anti‐E.
In vitro Evaluation of a Very‐High‐Purity, Solvent/Detergent‐Treated, Von Willebrand Factor Concentrate
Authors: Mazurier Claudine , Jorieux Sylvie , de Romeuf Christophe , Samor Bruno , Goudemand Maurice
Published: March 2009

Abstract. A very highly purified von Willebrand factor (vWF) concentrate was analyzed in order to evaluate its suitability for the treatment of von Willebrand's disease (vWD). The functional activity of vWF assessed by its ristocetin cofactor activity (vWF:RCo) correlated with the level of vWF antigen (vWF:Ag), with the vWF:RCo/vWF:Ag ratios ranging from 0.56 to 1.02, and the specific activity being always > 50 IU vWF:RCo/mg protein. Electrophoretic analysis showed a normal pattern of high, intermediate and low‐molecular‐weight multimers of vWF. The biological activity of vWF was also evaluated by studying its ability to bind to collagen and to platelet receptors in the presence of either ristocetin or thrombin. Furthermore, these functional activities of vWF were confirmed by the capacity of this concentrate to induce platelet adhesion to collagen in a perfusion system. Moreover, the vWF present in this preparation was able to bind factor VIII. All these in vitro data suggest that this preparation is likely to be effective in the treatment of vWD patients.
The in vitro and in vivo Evaluation of Whole Blood and Red Cell Concentrates Drawn on CPDA‐1 and Stored in a Non‐DEHP Plasticized PVC Container
Authors: Seidl S. , Gosda W. , Reppucci A.J.
Published: March 2009

Abstract.A new polyvinyl chloride (PVC) blood storage container, PL 2209, plasticized with butyryl‐n‐trihexyl‐citrate (BTHC) instead of diethylhexyl phthalate (DEHP) was used for in vitro and in vivo studies. Whole blood and red cell concentrates drawn on CPDA‐1 were stored 42 days at 1–6°C. Various biochemical parameters were tested and compared to those values seen in the literature for products stored in DEHP‐plasticized containers. Measurements of lactate production, glucose consumption, sodium and potassium ions for both whole blood and red cells were compatible with published data. Adenosine triphosphate (ATP) values were on the average 73% for the red cell units and 80% for the whole blood units after 35 days storage. Hemolysis was 0.43% for the red cell concentrate and 0.38% for the whole blood units after the same storage period. Autologous recovery studies after 35 days of storage for both the whole blood units and red cell concentrate gave 80% recovery 24 h after reinjection. From these data it can be concluded that PL 2209 BTHC‐plasticized PVC blood storage containers allow 35 days storage of blood draw on CPDA‐1 and can be considered as an alternative to DEHP‐plasticized PVC containers.
Development of Non‐Rh Antibodies in Volunteers Stimulated for the Production of Hyperimmune Anti‐D
Authors: Teesdale Phyllis , de Silva Mahes , Contreras Marcela
Published: March 2009

Abstract. At the North London Blood Transfusion Centre, red cells from accredited Rh‐D‐positive donors, matched for all antigens capable of inducing clinically significant antibodies, are used to stimulate immune plasma donors to achieve higher anti‐D levels. Despite such careful matching, antibody to the relatively non‐immunogenic M antigen developed in 3 out of 20 NN donors (15%) stimulated with M‐positive cells. In general, good responders to the Rh antigen D are good responders to other red cell antigens; our report exemplifies the importance of using fully matched accredited red cells for immune stimulation and the need to perform thorough antibody screening after each stimulation.
Antibody Removal Therapy Used Successfully at Delivery of a Pregnant Patient with Glanzmann's Thrombasthenia and Multiple Anti‐Platelet Antibodies
Authors: Ito Kazuhiko , Yoshida Hisahiro , Hatoyama Hiroshi , Matsumoto Hisashi , Ban Chiaki , Mori Takahide , Sugiyama Tateo , Ishibashi Takafumi , Okuma Minoru , Uchino Haruto , Maruya Etsuko , Oki Atsuko , Saji Hiroh , Hosoi Takemitsu , Tanoue Kenjiro , Tomiyama Yoshiaki , Kurata Yoshiyuki
Published: March 2009

Abstract. A 31‐year‐old Japanese woman with Glanzmann's thrombasthenia became pregnant voluntarily. She had had transfusions with more than 60 units for severe bleeding. She had multiple antibodies against HLA antigens and platelet glycoprotein Ilb/IIIa. No compatible platelets were available. To prevent serious hemorrhage during her delivery, antibody removal therapy was carried out three times. Large molecules including immunoglobulins were removed from more than 3 liters of plasma each time. After the titer of antiplatelet antibodies had decreased in the patient's blood, antihuman globulin‐lymphocyte cytotoxicity test compatible platelets were transfused. Her bleeding time improved and delivery was induced successfully despite atonic hemorrhage of about 2,000 g of blood. Her infant had no bleeding problems. This patient is the first with Glanzmann's thrombasthenia to receive antibody removal therapy at delivery.
An Improved Method to Study Complement Receptor‐Mediated Function of the Fixed Macrophage System in vivo
Authors: Ekdahl Kristina Nilsson , Lööf Lars , Nilsson Ulf R. , Nilsson Bo
Published: March 2009

Abstract. A method to coat unsensitized erythrocytes with fragments of C3 and C4 using autologous serum, in order to study complement receptor‐dependent function of the fixed macrophage system, is presented. After incubation with serum under optimal conditions, at least 90% of the cells had C3b/iC3b deposited on the surface, with an average of 20 × 103 molecules per cell. Elimination of the coated cells by the fixed macrophage system was studied in 12 normal subjects. With a dose of 4.5 × 108 red cells injected, 75% of the cells were eliminated with a half‐life of approximately 2.4 ± 0.3 min (n = 7). In subjects receiving ten times more cells, there was a rapid decrease in the amount of C3‐coated cells, reaching a nadir with 85% remaining for 4–6 min, after which there was a gradual release of cells for another 20 min (n = 5). In absolute numbers, 3 × 108 of labeled cells were eliminated regardless of the dose injected. The coating procedure presented here is simple, does not introduce heterologous blood components and makes it possible to control the amount and the degree of fragmentation of the C3 and C4 deposited on the erythrocyte surface.
A New Monoclonal Anti‐A Antibody BIRMA‐1
Authors: McDonald D. F. , Thompson J. M.
Published: March 2009

Abstract. A monoclonal anti‐A antibody, BIRMA‐1, has been evaluated and found to be eminently suitable as a blood‐grouping reagent. The culture supernatant is potent, avid and specific as demonstrated by its reactivity with a large sample of donor bloods tested manually and on the Olympus PK 7100 automated blood grouping machine. Over 100 cord blood samples were tested manually, and all were correctly grouped using this antibody. No false‐positive reactions were obtained with papainized group B cells. Reagent prepared from BIRMA‐1 detected most examples of Ax and all other A subgroups and A variants tested. Negative reactions with B(A) cells indicate that it is possible to have an antibody capable of detecting Ax which will not react with B cells from individuals with high levels of galactosyltransferase.
Aluminum Content of Source Plasma and Sodium Citrate Anticoagulant
Authors: May Joan C. , Etz Nora M. , Rains Theodore C. , Yu Li Jian
Published: March 2009

The concentration of aluminum was determined in samples of source plasma collected by the normal plasmapheresis procedure, which involves collection in anticoagulant and immediate freezing. Samples of sodium citrate anticoagulant used in the collection of source plasma were also tested for aluminum, as were empty source plasma containers and 0.9% sodium chloride infusion (USP). Samples of source plasma were collected from a geographic cross‐section of the donor population in the USA by three different manufacturers. Aliquots of these samples were mixed with Triton X‐100 and sulfuric acid and analyzed for aluminum by atomic absorption spectrometry using electrothermal atomization (graphite furnace) and Zeeman background correction. The arithmetic mean and standard deviation for the aluminum content of 28 samples of source plasma were found to be 25.5 ± 8.4 ng Al/ml. The aluminum content of the individual samples of source plasma ranged from 12 to 48 ng Al/ml. The aluminum content of 6 samples from two manufacturers of the sodium citrate anticoagulant that is used in source plasma ranged from 410 to 2,080 ng/ml. Aluminum levels found in saline for infusion and nitric acid leachates from empty source plasma containers were less than 6.9 ng/ml. The level of aluminum expected in uncontaminated human blood has been estimated to be 10 ng Al/ml or less. Comparison of this figure with the present data indicates that the sodium citrate anticoagulant contributes significantly to the aluminum load of source plasma and, therefore, to the aluminum content of products such as albumin derived from source plasma.
Evaluation of Four Different Methods for Platelet Freezing: In vitro and in vivo Studies
Authors: Angelini Antonio , Dragani Alfredo , Berardi Anna , Iacone Antonio , Fioritoni Guiseppe , Torlontano Glauco
Published: March 2009

This report describes our experience with various techniques for the freezing of platelet‐rich plasma, removed from the final product after leukapheresis procedures performed on 14 hematological patients. A total of 194 platelet units were frozen for subsequent autologous transfusion, by the following four methods: (1) 6% dimethyl sulfoxide (DMSO); (2) a combination of 5% DMSO/6% hydroxyethyl starch; (3) 3% glycerol; (4) 5% glycerol/4% glucose. Each technique was evaluated by measuring the percentage of platelet recovery, malondialdehyde (MDA) production, and lactate dehydrogenase release. To investigate the safety and therapeutic effectiveness of the previously frozen platelets, in vivo comparison of four platelet freezing methods was made in 8 thrombocytopenic patients, using corrected platelet increment (CCI), determined at 24 h. Our in vitro results indicate that the cryopreservation with 6% DMSO, without controlled cooling rate, provides significantly (p < 0.05) greater platelet recovery (75%) as compared to other systems. The decrease of MDA production and the increase in plasma lactate measured after the thawing process was less in the DMSO‐frozen units than in the other platelet units. When platelets, cryopreserved by this method, were subsequently transfused into patients, a significantly better CCI (> 5,000/μl) was obtained. In our series, 6 patients were entirely supported with frozen autologous platelets. It appears from this study that a better understanding of the physical and biochemical events occurring during the freezing process will improve platelet cryopreservation, allowing a more systematic use of frozen platelets in the support of thrombocytopenic patients.
Haemolytic Disease of the Newborn Probably due to Anti‐ELO, an Antibody to a Low Frequency Red Cell Antigen
Authors: Ford D. S. , Stern D. A. , Hawksworth D. N. , Better P.J. , Lubenko A. , Pope J. M. , Ghana H. S.
Published: March 2009

Investigation of mild bilirubinaemia and a positive direct antiglobulin test in a 2 day old baby revealed that the mother's serum contained an antibody against the low frequency antigen ELO, which was present on the father's red cells. Family studies showed that the ELO antigen segregates from Rh, Gc, ADA and PGM. The ELO antigen is enzyme resistant and therefore not likely to be part of the MNS or Duffy systems. No abnormalities were detected in immunoblotting studies. Although insufficient samples were available to attempt elution of the antibody from the cord cells, it is probable that this case represents the first reported haemolytic disease of the newborn due to anti‐ELO.
Anti‐IgG Autoantibodies in HIV‐infected Hemophilia Patients
Authors: Süsal C. , Lewin I. V. , Stanworth D. R. , Terness P. , Daniel V. , Oberg H.‐H. , Huth‐Kuhne A. , Zimmermann R. , Opelz G.
Published: March 2009

Sera of 76 HIV‐negative hemophilia patients, 103 HIV‐positive (HIV+) hemophilia patients free of AIDS or AIDS related complex (ARC), and 32 HIV+ hemophilia patients with AIDS/ARC were tested for four different anti‐IgG activities. IgG‐anti‐F(ab')2γ, IgM‐anti‐F(ab')2γ, and IgG‐anti‐Fcγ serum activities were significantly associated with the clinical stage of HIV infection, whereas IgM‐anti‐Fcγ was not. IgG‐anti‐F(ab')2γ activity was found to be caused by cross‐reaction of anti‐HIV antibody with an epitope within the constant CHI domain of human IgG. HIV+ hemophilia patients with severe thrombocytopenia (< 50,000/μl platelet counts) had significantly higher IgM‐anti‐IgG activity than patients with > 50,000/μl platelets. Because anti‐IgG antibodies possess immunoregulatory properties, our results may serve as a possible explanation for the frequent B cell disorders encountered in HIV‐infected patients.
Virus Inactivation during Intravenous Immunoglobulin Production
Authors: Hämäläinen E. , Suomela H. , Ukkonen P.
Published: March 2009

Effects of time, temperature, pH and stabilizers (i.e. medium) on inactivation of lipid‐enveloped model viruses, Semliki Forest and vesicular stomatitis viruses in the production process of intravenous immunoglobulin were investigated on a laboratory scale. The lowering of pH, the raising of temperature and the increasing of incubation time improved the inactivation effect. However, small changes in pH and stabilizer concentrations did not influence the results. Inactivation was not linear and a clear tailing off could be seen. Therefore, for complete virus inactivation incubation times longer than 20 h are necessary. Inactivation took place much more rapidly in intravenous immunoglobulin solution than in intramuscular immunoglobulin solution. Processing steps such as freeze‐dying in the presence of ethanol or storage of intramuscular immunoglobulin in the liquid state at pH 7 only partially inactivated these viruses.