bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro and In vivo Evaluation of Platelet Concentrates after Cotton Wool Filtration
Authors: Holme S. , Ross D. , Heaton W. A.
Published: March 2009

Abstract. Removal of leukocytes from platelet concentrates (PC) may decrease the incidence of alloimmunization to HLA antigens on white cells and prevent or delay refractoriness. Cotton wool filtration, which effectively removes leukocytes from PC with minimal platelet loss, may cause platelet or complement activation. In this study, the effect of cotton wool filtration (Imugard® IG‐500) on platelet activation, in vitro function, posttransfusion survival, and complement activation was investigated. Five paired autologous in vivo percent recovery and survival studies were performed with fresh (6–8 h) PC or with 5‐day‐stored PC prepared from CPD‐anticoagulated blood and stored in PL‐732TM containers using standard methods. In the paired design, PC from the same donor were filtered on one occasion, and not filtered on another, prior to labeling with indium‐111‐oxine and reinfusion. Percent recovery and survival were then determined by the standardized method. Filtration had no statistically significant effect on percent recovery on PC stored for 6–8 h or for 5 days. There was, however, a slight decrease in survival hours of the filtered PC which was statistically significant at 6–8 h (203±14 vs. 179±18 h; p<0.05) but not at 5 days (166±28 vs. 132±27 h; p>0.05). Samples taken before and after filtration of single units (fresh and 5‐day) and pooled units (3‐day) for measurement of release of granular content (β‐thromboglobulin), lysis (LDH), and in vitro viability—ATP, extent of shape change with ADP, and hypotonic shock response (3‐day only)—demonstrated no effect of filtration on these parameters. A slight, but significant (p<0.05) increase in plasma levels of C3a was found for fresh‐filtered PC, but not for stored PC, suggesting some complement activation during filtration.
Storage of Whole Blood for up to 24 Hours at Ambient Temperature prior to Component Preparation
Authors: Pietersz R.N.I. , de Korte D. , Reesink H. W. , Dekker W.J.A. , van den Ende A. , Loos J.A.
Published: March 2009

Abstract. The effect of rapid cooling to 20–24 °C of whole blood immediately after collection, using ‘cooling units' with butane‐1,4‐diol and prolonged storage up to 24 h at ambient temperature was investigated in the whole blood and the subsequently prepared plasma, buffy coat and buffy‐coat‐poor red cell concentrate (BC‐poor RCC) in saline‐adenine‐glucose‐mannitol (SAGM) solution. Factor VIII:C content of the plasma (n=10), after 24 h storage was 80± 3% of the initial value. In routine procedures factor VIII:C content in the plasma (n= 129 pools of 20 donor units plasma) was 0.77 ± 0.078 IU/ml, after storage of the whole blood for 16–20 h. In whole blood (n=10), the 2,3‐diphosphoglycerate (2,3‐DPG) content of the red cells decreased from 4.36 ± 0.55 to 1.47 ± 0.6 μmol/ml red cells after 24 h storage at 20–24°C. After storage of the BC‐poor RCC (n=10) at 2–6°C for 1 week, the 2,3‐DPG had dropped to 0.76 ± 0.46 μmol/ml red cells. During the first 24 h of storage of whole blood, the adenine triphosphate (ATP) levels of the red cells remained stable. A mean increase of 20% of the initial value was observed after addition of SAG M solution. In the BC‐poor RCC the ATP slowly decreased to 81 ± 5% after 5 weeks and to 68 ± 6.6% of the initial value after 6 weeks storage. In citrate‐phosphate‐dextrose blood the yield of platelets in the buffy coat was found to be 84 ± 6% (mean ± SD) of the original value when whole blood (n= 12) was stored for 16–20 h at 20–24 °C, as compared to 76 ± 18%, when buffy coats were prepared within 3 h after collection of whole blood without rapid cooling (n= 12). Rapid cooling of whole blood to 20–24 °C immediately after collection and subsequent storage of the whole blood up to 24 h contributes to the quality and standardization of the subsequently prepared blood components and will diminish processing at irregular hours.
Prevention of Recurrent Spontaneous Abortion by Intravenous Immunoglobulin
Authors: Mueller‐Eckhardt Gertrud , Heine Olaf , Neppert Jürgen , Künzel Wolfgang , Mueller‐Eckhardt Christian
Published: March 2009

Abstract. Intravenous immunoglobulin (IVIG) treatment was attempted as a novel therapeutic approach for unexplained recurrent spontaneous abortions (RSA) occurring in the first trimester of pregnancy. Twenty women with a history of RSA were treated with IVIG during pregnancy. Therapy was commenced at week 5 of gestation with 1 dose of 0.5–0.6 g IVIG/kg body weight. Infusions were repeated every 3 weeks (0.3–0.4 g/kg) and terminated by week 22 to 24. Of 20 women, 11 delivered healthy infants at term. 5 women are still pregnant, 3 in the third trimester. Only 3 patients suffered abortions and 1 presented with ectopic pregnancy. The overall success rate was 82–86%. Thus, the therapeutic effect of IVIG is comparable to that of the conventional transfusion/vaccination regimen with allogeneic leukocytes, but avoids the risk of transmission of infections and/or HLA immunization, has no major adverse effects and is applicable to ‘nonresponders’.
Immunoglobulin Classes of Antibody for Human T‐Lymphotropic Virus Type‐I (HTLV‐I) in Healthy Donors and HTLV‐I‐Associated Disorders
Authors: Kamihira Simeru , Sohda Hisasi , Oyakawa Nobuyuki , Moriuti Yukiyosi , Momita Saburo , Ikeda Shuiti , Yamada Yasuaki , Ichimaru Michito , Kinosita Kenichirou , Okuda Hiroto , Kanamura Machiko , Oota Takiko
Published: March 2009

Abstract. Healthy blood donors, patients with adult T‐cell leukemia (ATL) and HTLV‐I‐associated myelopathy (HAM) and recipients of unscreened blood (SR) who had seroconverted and were followed‐up for more than 2 years were examined for HTLV‐1 antibodies of immunoglobulin G (IgG) and M(IgM) classes. The overall infection rate in donors was 4.9%, as determined by screening with a particle agglutination method (PA). The rate increased with increasing age. Positive sera with a low titer in the PA test (1/16, 1/32 and 1/64) contained IgM antibodies in 32.5% (titer 1/16) to 36.1% (titer 1/64) of the cases, but IgG antibodies were detected in only 5.6% of the sera with a titer of 1/16 and in 36.1% of the sera with a titer of 1/64. Conversely, in high titer sera (1/128 or higher) IgG antibodies were almost always detectable (99.0%) and IgM antibodies less frequently (25.5%). Sera from acute, chronic and pre‐ATL, HAM and SR patients contained IgG antibodies in high titer in all cases. The incidence of IgM antibodies was 7.7, 30.0, 53.3, 72.3, and 77.8%, respectively. IgM antibodies were demonstrated repeatedly in some cases who were followed up for a year. Only IgM antibodies from HAM patients occurred in high titers and had strong reactivity to the p24 antigens of HTLV‐I in Western blot testing. It is concluded that it is important to detect IgM antibodies not only in primary infections but also in persistent infections of HTLV‐I.
A Novel Method for Removal of Human Immunodeficiency Virus: Filtration with Porous Polymeric Membranes
Authors: Hamamoto Yoshiaki , Harada Shinji , Kobayashi Susumu , Yamaguchi Kazuhito , Iijima Hideki , Manabe Sei‐ichi , Tsurumi Takashi , Aizawa Hiizu , Yamamoto Naoki
Published: March 2009

Abstract. We propose a new method to rid solutions of a virus by using a novel regenerated multilayered structured cellulose membrane (BMM). When the filtrate of human immunodeficiency virus (HIV) preparation was obtained through BMM it showed no infectivity. Electron microscopic observation revealed that HIV was completely caught by the multilayers of the BMM. Conveniently, BMM was seldomly found to adsorb protein molecules and also to have a high filtration rate. These characteristics may have a use in the removal of other variously sized pathogenic agents from plasma.
Serological and Immunological Characteristics of Maternal Anti‐Rh(D) Antibodies in Predicting the Severity of Haemolytic Disease of the Newborn
Authors: Żupańska B. , Brojer E. , Richards Y. , Lenkiewicz B. , Seyfried H. , Howell P.
Published: March 2009

Abstract. A number of factors were analyzed for their predictive value in indicating the severity of haemolytic disease of the newborn (HDN) in 72 infants. The factors investigated were: maternal antibody titre in the indirect antiglobulin test, the number of antibody molecules on sensitized standard red cells evaluated by a radiometric antiglobulin test, the IgG subclass specificity and the reactivity in monocyte‐monolayer assay (MMA) and in the rosette assays with lymphocytes and granulocytes from healthy individuals. The results of the MMA correlate much better with the severity of HDN than the antibody titre. In clinically unaffected infants the reactivity in the MMA never exceeded 20%, while in the severe/very severe group it was always greater than 20% (in 95% of very severe cases even above 50%). The number of IgG‐bound molecules was also shown to closely correlate with the clinical severity and there was a much greater proportion of severe/very severe cases exhibiting combined IgG1 and IgG3 specificity. Of all the evaluations performed the rosette assays with lymphocytes and granulocytes were found to be less useful in predicting the severity of HDN.
Testing Efficacy of Anti‐D Sera by a Panel of Donor Red Cells with Weak Reacting D Antigen and with Partial D Antigens
Authors: van Rhenen D.J. , Thijssen P.M.H.J. , Overbeeke M.A.M.
Published: March 2009

Abstract. In 1987 the definition of an Rh‐negative donor in the Netherlands was changed from C‐, E‐ as well as Du‐negative to D‐negative only. The use of 2 different strong anti‐D sera without an antiglobulin phase (Du test) was considered sufficient to reveal the clinically important D antigen. In applying this policy, we identified 32 donors in 13,500 consecutive blood donations whose indirect antiglobulin test (IAT) (Du)‐positive red cells gave negative reactions with at least 2 out of 11 anti‐D sera and whose cells might therefore be typed as Rh(‐D‐)‐negative in routine investigations. IgG anti‐D used by a one‐stage bromelain technique and anti‐D with modified IgG appeared to be relatively insensitive in detecting Du in this study. Polyclonal anti‐D in an enhancement medium and monoclonal anti‐D scored better, although differences exist between the products of various manufacturers. It is suggested that if IAT (Du) testing is omitted, only anti‐D sera with a high index of detectability of cells expressing weak D antigens should be accepted.
A New Cellulose Acetate Filter to Remove Leukocytes from Buffy‐Coat‐Poor Red Cell Concentrates
Authors: Pietersz R.N.I. , Dekker W.J.A. , Reesink H.W.
Published: March 2009

Abstract. Transfusion of leukocyte‐free red cell concentrates (RCC) prevents or delays HLA immunization in multitransfused patients. We investigated a new cellulose acetate filter which was recently introduced to remove leukocytes from buffy‐coat‐poor RCC. It was found that the filtration time was only 10 min with buffy‐coat‐poor RCC in saline‐adenine‐glucose‐mannitol (SAG M; n=23), hematocrit being 62 ± 2% (SD). The red cell loss was 13.5 ± 2.6% and leukocyte removal was more than 99%. Routine filtration in SAG M (n=179) showed again that more than 99% of the leukocytes were removed from buffy‐coat‐poor RCC with an original leukocyte content of 804 ± (SD)458times 106. The red cell loss (12 ± 8.6%) was not diminished by increasing the amount of saline (0.9% NaCl) from 100 to 300 ml in an attempt to remove the retained red cells from the filter. We conclude that the new filter is reliable in rapidly removing more than 99% of the leukocytes from buffy‐coat‐poor RCC in SAG M solution.
The Fine Specificity of Lewis Blood Group Antibodies. Evidence for Maturation of the Immune Response1
Authors: Cowles J. W. , Spitalnik S. L. , Blumberg N.
Published: March 2009

Abstract. Eight human Lewis blood group antibodies were characterized for their fine specificity by the use of specific immunoadsorbents and a kinetic enzyme‐linked immunosorbent assay technique. Examination of sera following immunoglobulin fractionation showed IgM anti‐Lea exhibiting broad cross‐reactivity with structures biochemically related to the Lewis antigens. IgG anti‐Lea binding was restricted to Lea and Leb. These findings are consistent with the concept of affinity maturation of the immune response, which has been previously demonstrated only in animal model systems.
Stability of Murine Monoclonal Anti‐A, Anti‐B and Anti‐A,B ABO Grouping Reagents and A Multi‐Centre Evaluation of Their Performance in Routine Use1
Authors: McGowan A. , Tod Anne , Chirnside Ann , Green C. , McColl K. , Moore S. , Yap P.L. , McClelland D.B.L. , McCann Moira C. , Micklem Lisel R. , James K.
Published: March 2009

Abstract. We have previously reported the production of 3 murine monoclonal reagents for ABO typing (designated ES‐9, ES‐4 and ES‐15). This study presents results of tests of stability of these 3 reagents, together with a fourth murine monoclonal antibody (LM103/107). In addition, data are also presented from a multi‐centre evaluation of the performance of the murine monoclonal reagents in routine ABO typing of both donors and patients using a wide variety of techniques, both manual and automated. The potency and stability of the 4 monoclonal antibody based reagents is compared with a broad selection of monoclonal and polyclonal ABO typing reagents. The reagents used for comparison were produced by European and United Stated manufacturers in both the public and private sector and are widely used in routine ABO typing. The Scottish monoclonal reagents have been used successfully to ABO type over 500,000 blood samples in 7 centres within the UK, with no discrepant results.
Absence of HIV DNA Sequences in Seronegative Polytransfused Thalassemic Patients
Authors: Lefrère Jean‐Jacques , De Montalembert Mariane , Mariotti Martine , Girot Robert , Salmon Charles , Rouger Philippe , Rey Jean
Published: March 2009

The risk of infection with human immunodeficiency virus (HIV) by transfusion is not totally eliminated, since contaminated blood given before seroconversion to HIV is not detected on the actual biological screening. We used the polymerase chain reaction (PCR) assay (with one primer pair in the gag region and two in the pol region) to detect HIV DNA sequences in 30 seronegative polytransfused thalassemic patients and in 60 seropositive individuals (used as positive controls). We did not observe PCR‐positive HIV‐antibody‐negative results in seronegative polytransfused patients.
Effects of Hypertonic Saline (7.5%)/Dextran 70 on Human Red Cell Typing, Lysis, and Metabolism in vitro1
Authors: Moore Gerald L. , Summary James J. , Dubick Michael A. , Ledford Mary E. , Ryan Barbara A. , Gonzales Armando , Wade Charles E.
Published: March 2009

The introduction of a 7.5% hypertonic saline/6% dextran 70 (HSD) solution into clinical trials for the treatment of hypovolemic states, and the past concerns regarding the possible interference of dextran with blood serology, prompted us to investigate the effects of HSD on human red‐cell typing and stability. HSD was evaluated with fresh and 35‐day stored CPDA‐1 red cells from 12 healthy donors. A 1:5 mixture of HSD to blood in vitro had no effect on ABO, Rh, and MN typing in both fresh and stored blood. HSD produced no significant lysis with fresh cells and a minimal level with stored blood. No evidence of metabolic or morphologic changes was seen after HSD treatment. The results of this study suggest that the clinical use of HSD for the treatment of hemorrhagic shock will not affect blood group determinations or red‐cell stability from stored blood which may be infused after the HSD‐treated patient is transported to a hospital.
A New, Low‐Incidence Red Cell Antigen (HOFM), Associated with Depressed C Antigen
Authors: Hoffmann J.J.M.L. , Overbeeke M.A.M. , Kaita H. , Loomans A.A.H.
Published: March 2009

A case of mild hemolytic disease of the newborn is presented which was caused by an antibody to a hitherto unknown antigen of low incidence. This antigen, now designated as HOFM (ISBT number 700050) was detected in 6 relatives, and in all of them, it was associated with an unusually weakened expression of C antigen. The serological data indicate that HOFM may be part of the Rh system, but the genetic data, although supportive of this interpretation, are inconclusive.
Development of an Immunoaffinity Process for Factor IX Purification
Authors: Tharakan John , Strickland Dudley , Burgess Wilson , Drohan William N. , Clark David B.
Published: March 2009

Abstract. An immunoaffinity process based on monoclonal antibody (MAb) to factor IX (FIX) has been developed. Initially, vitamin‐K‐dependent proteins from cryoprecipitate‐poor plasma are isolated on DEAE‐Sephadex. The eluate is applied to an immunoaffinity column that utilizes a divalent metal‐ion‐dependent MAb directed against FIX. After washing the column with high salt in the presence of magnesium ion, the FIX is eluted using a citrate‐ or EDTA‐containing buffer. Coagulation assays and Western blots show no detectable amounts of any contaminating proteins. Purity of the FIX product is established using reduced and nonreduced Coomassie‐stanined SDS‐PAGE and HPLC. The N‐terminal 20 amino acids of the single peak of the HPLC were shown to be identical to those reported for FIX. The process shows no detectable leakage of monoclonal antibodies (MAb), efficient utilization of MAb, and provides yields greater than 95%. The use of solvent/detergent treatment as a potential viral inactivation method is incorporated in the process. Studies with tritiated Triton X‐100 indicate that the detergent can be washed out of the MAb column so that less than 1 ppm (total) Triton X‐100 coelutes with the FIX.
Studies in Red Blood Cell Preservation 2. Comparison of Vesicle Formation, Morphology, and Membrane Lipids during Storage in AS‐1 and CPDA‐1
Authors: Greenwalt T. J. , Sostok C. Zehner , Dumaswala U. J.
Published: March 2009

Abstract. The changes in morphology, the quantitative changes in membrane lipids and the shedding of exocytic vesicles by red blood cells (RBC) stored for 42 and 56 days in AS‐1 and CPDA‐1 were compared. RBC stored in AS‐1 shed significantly less vesicle membrane cholesterol, phospholipid and protein and maintained better morphology scores. RBC membrane cholesterol remained higher after 56 days in AS‐1 than in CPDA‐1. The data suggest that during the first weeks of storage cholesterol is lost from the RBC membrane followed by a larger release of phospholipids accompanied by alterations in the phosphoinositides. The shedding of exocytic vesicles appears to be secondary to the changes in morphology resulting from the perturbation of the membrane lipids.
Platelet Collection with the Autopheresis‐C® Apheresis System
Authors: Schoendorfer D. W. , Williamson L. H. , Sheckler V. L. , Fitzgerald B. P.
Published: March 2009

Abstract. This report describes a new system for collection of platelet concentrate (PC) and cell‐free plasma (PPP) from apheresis donors. The system uses two separation devices and requires only a single venipuncture. The Plateletcell™ device separates primary platelet concentrate (PPC) from anticoagulated whole blood and the Plasmacell‐C® device separates the PPC into PC and PPP. Results of functional studies performed indicate that the separation process does not alter viability of either the PPC, the PC, or the PPP. Platelet function after 5 days of storage is maintained. An average yield of 3.4±0.7 × 1011 platelets in 201 g of PC and 422 g of PPP were harvested in 71 ±13 min of donor time from donors with preprocedure hematocrits averaging 42.5±2.0% and preprocedure platelet counts averaging 265±61 times 103/μl.
Massive Transfusion in Adults:Diagnoses, Survival and Blood Bank Support
Authors: Sawyer P. R. , Harrison C. R.
Published: March 2009

Abstract. In a retrospective study covering a 2‐year period (1986–1987), 125 patients received massive blood transfusion. Trauma accounted for only 29% of the cases. The larger nontrauma diagnostic categories were: gastrointestinal hemorrhage, 31%; cardiovascular surgery, 12%, and oncology cases, 9%. The overall survival rate was 60%; survival rates ranged from 38% for patients with hepatic failure to 100% for obstetric cases. Clinically important alloantibodies were present in 4%, and transfusion reactions occurred in 9%. Massive transfusions accounted for a significant proportion of total blood component usage: at least 12% of the yearly total red cell units, 20% of plasma, and 14% of platelets transfused.
Nonradioactive Chromium Technique for Red Cell Labeling
Authors: Sioufi H. A. , Button L. N. , Jacobson M. S. , Kevy S. V.
Published: March 2009

Abstract. Following transfusion, red cells labeled with nonradioactive chromium (52Cr) can be detected in the circulation using atomic absorption spectrophotometry. Both in rhesus monkeys and in human subjects, the survival of red cells labeled with 52Cr was found to be insignificantly different from that of cells labeled with 51Cr. Nonradioactive chromium can be used to label a second population of red cells, when 51Cr is used for the first population, or can be used on its own when the use of radioactivity is contraindicated.
Evaluation of the Polyethylene Glycol Antiglobulin Test for Detection of Red Blood Cell Antibodies
Authors: de Man A.J.M. , Overbeeke M.A.M.
Published: March 2009

Abstract. The polyethylene glycol (PEG) antiglobulin test (AGT) was evaluated by testing antibodies in the sera of 362 different patients and by screening of 4,685 random patient sera with both the PEG AGT and the albumin AGT. Agglutination in the PEG method was stronger in 254 (70.2%), identical in 99 (27.3%) and weaker in 9 (2.5%) of the 363 sera. Mean end‐point titers of the antibodies measured with the PEG method (n = 137) were 2–3 dilutions higher than with the albumin method. In sera with anti‐K and anti‐Fyb antibodies the mean titer in the PEG method was only one dilution higher. Antibodies within the rhesus system were also tested with a two‐stage enzyme test. Agglutination in the PEG method was stronger in 93 (50.3%), identical in 64 (34.6%) and weaker in 28 (15.1%) of the 185 sera. Mean antibody titers (n = 76) in the PEG method were 1–3 dilution steps higher for the various specificities than in the enzyme test. In 92 (2.0%) of the 4,685 sera screened in the PEG method, specific antibodies were detected; with the albumin method in 74 (1.6%) sera. The antibodies only detected with the PEG method had the specificities anti‐D, ‐C, ‐Cw, ‐c, E, ‐K, ‐Jka and ‐M. Not detected by the PEG method were anti‐Lea antibodies in one of the sera. It is concluded that the PEG AGT is a sensitive technique for the detection and identification of red cell antibodies and that it is superior to the albumin AGT.
Prophylactic Intravenous Immunoglobulin in Pre‐Term Infants: A Controlled Trial
Authors: Conway Steven P. , Ng Pak C. , Howel Denise , Maclain Bruce , Gooi Hock C.
Published: March 2009

Abstract. Sixty‐six pre‐term infants of less than 30 weeks gestation consecutively admitted to either of two neonatal intensive care units were randomized to receive routine intensive care only or prophylactic intravenous immunoglobulin, 200 mg/kg body weight at 3‐weekly intervals in addition to routine intensive care. Eleven babies, 6 in the control group and 5 in the treatment group, were withdrawn from the trial due to early death from extreme prematurity (7 babies), early return to the referring hospital (3 babies), and elective treatment with intravenous immunoglobulin for severe congenital septicaemia (1 baby). Of the 55 who remained in the trial, significantly fewer babies in the treatment group had infection, 8 compared to 17 (p = 0.01). This difference was not significant when blood‐culture‐proven septicaemia only was considered, 8 compared to 14 (p = 0.09). Twenty‐seven (84%) of 32 blood‐culture‐positive episodes of infection were caused by coagulase‐negative staphylococci. Serum IgG was significantly higher with treatment and achieved levels comparable to those of normal full‐term babies at the same post‐natal age.
Systematic Pre‐Deposit Autologous Blood Provision for Elective Surgery: An Important Contribution to Hospital Blood Supply
Authors: Kay L.A. , Noble R.S.
Published: March 2009

Abstract. From the 8th of September 1987 to the 31st of July 1988 all patients scheduled for major elective orthopaedic surgery were systematically offered the facility of pre‐deposit autologous blood storage. Of 251 patients, 204 (81%) were eligible to pre‐donate, and of these only 9 declined to do so. 72% of the participants required only autologous blood at operation, 23% required some additional homologous blood. During the study period, autologous blood accounted for 10% of all blood transfused in the district, making a significant contribution to the local blood supply.
The Red Cell Antigen JAL in the Swiss Population: Family Studies Showing That JAL Is an Rh Antigen (RH48)
Authors: Poole J. , Hustinx H. , Gerber H. , Lomas C. , Liew Y.W. , Tippett P.
Published: March 2009

Abstract. The JAL antigen was found to have an overall frequency of 0.004% in the Swiss population and 0.06% in French‐speaking Swiss. Family studies of 5 JAL+ individuals have shown that the JAL antigen is not part of the ABO, MNSs, Fy, Jk and Co blood group systems, or the Se system, nor is it X‐ or Y‐linked. JAL is encoded by the RH locus or by a very closely linked locus. The number RH48 (4.48) has been assigned for JAL by the International Society of Blood Transfusion Working Party on Terminology for Red Cell Surface Antigens.
Large‐Scale Gel Filtration Chromatography for the Production of a Solvent/Detergent‐Treated High‐Purity Factor VIII Concentrate
Authors: Dengler T. , Stöcker U. , Kellner S. , Fürst G.
Published: March 2009

Abstract. To produce a tri(n‐butyl)phosphate/sodium‐cholate‐treated intermediate‐purity factor VIII (FVIII) concentrate with a specific activity of about 1 IU/mg, we used a simple gel filtration step with Sephadex G25 to remove the solvent/detergent reagents from the final product. By exchanging the Sephadex G25 gel for a new high‐resolution gel (Sephacryl S400 HR), we obtained a high‐purity FVIII concentrate, by simultaneous elimination of about 98% of the extraneous proteins and removal of the solvent/detergent reagents, without reducing the FVIII:c yield and without altering the production scheme. With different protein analysis techniques we analysed the resulting FVIII concentrate and, comparing it to the formerly produced intermediate‐purity FVIII concentrate, demonstrated improved purity.
Correlation of Monocyte‐Monolayer Assay Results, Number of Erythrocyte‐Bound IgG Molecules, and IgG Subclass Composition in the Study of Red Cell Alloantibodies Other Than D
Authors: Zupańska B. , Brojer E. , McIntosh J. , Seyfried H. , Howell P.
Published: March 2009

Abstract. Comparisons have been made between the serological and immunological characteristics of 42 blood group allonantibodies (other than D) covering twelve systems using a monocyte‐monolayer assay (MMA), a radiometric antiglobulin test for antibody binding and IgG subclass determinations. The results of the MMA correlated well with the level of IgG molecules bound on incompatible cells, and the highest levels in both cases were associated with the presence of the IgG3 subclass. However, limited clinical data shows that, while in general the MMA clearly identifies the clinically significant antibodies, the correlation with the degree of clinical outcome is less well defined, and in some instances other factors may be operating to ameliorate the in vivo effect of the antibody.
Screening Plasma Donors for High‐Titre Antibody to Cytomegalovirus Using a Latex Agglutination Test
Authors: McDonald C. , Barbara J.A.J. , Al‐Izzi A. , Contreras M.
Published: March 2009

Abstract. Cytomegalovirus (CMV) can cause severe morbidity in immunosuppressed patients. Regional transfusion centres in the UK are required to supply high‐titre anti‐CMV plasma to the Blood Products Laboratory (BPL), now called ‘Bio‐Products Laboratory’, for the production of specific intravenous immunoglobulin at the Protein Fractionation Centre in Scotland. For this purpose, 703 plasmapheresis donors were screened by a modified latex agglutination test to assess their suitability as donors with high‐titre anti‐CMV. CMV antibodies were found in 48% of the donors. Seropositivity increased with age ranging from 33% in the 20‐ to 29‐year age group to 57% in the 40–49 age group, although the 50–59 age group showed a slight decline to 53%. In all age ranges except the 40–49 group, seropositivity was highest in the female population. With the latex test, 14% of the total donors screened had a titre ≥1 in 64, 7% ≥1 in 128 and 4.2% ≥1 in 256. Samples from 18 donors with titres ≥1 in 128 were sent to BPL and all samples were found to have a sufficiently high titre for the production of CMV immune plasma. For BPL the ‘cut‐off’ level for CMV immune plasma is a titre ≥1 in 64 by complement fixation. Since the establishment of a panel of donors with high‐titre anti‐CMV plasma, more than 10 kg of plasma are despatched monthly from our centre to BPL.
Serological Characterization of Murine Monoclonal Antibodies Directed against Acquired B Red Cells
Authors: Janvier D. , Veaux S. , Reviron M. , Guignier F. , Benbunan M.
Published: March 2009

Abstract. Balb/c mice were immunized with acquired B red cells. Twelve clones specific for acquired B red cells were obtained from two fusions. A detailed investigation of three clones is reported here. These antibodies appear to be directed to the B‐like epitope since they are inhibited by galactosamine and fail to react after acetylation of red cells. E 231 is an example of a series of antibodies closely specific for acquired B red cells which can be useful in elucidating some AB0 typing problems. E 167 and F 47 showed a cross‐reactivity with A1 red cells and a synthetic A trisaccharide. No affinity for B antigen could be demonstrated for any of the antibodies.
Quantitation of Soluble HLA Class II Molecules by an Enzyme‐Linked Immunosorbent Assay
Authors: Westhoff Ulrike , Thinnes Friedrich P. , Götz Hilde , Grosse‐Wilde Hans
Published: March 2009

In order to quantify soluble HLA‐DR,DQ,DP molecules (sHLA‐RQP) an enzyme‐linked immunosorbent assay was developed utilizing two monoclonal antibodies specific for HLA‐DR,DP (Tü35) and HLA‐DQ (Tü22) gene products, respectively. Highly purified HLA class II molecules isolated from a lymphoblastoid cell line were used for calculation of exact sHLA‐RQP protein values. Circadian variations of sHLA‐RQP plasma levels were studied in 7 healthy probands showing no significant deviations; measurements in 4 probands at intervals between 4 and 6 weeks revealed that sHLA‐RQP levels remain relatively stable. The population analysis of 209 unrelated, HLA‐typed healthy donors resulted in an average protein concentration of 1.53 ± 2.44 μg/ml plasma for sHLA‐RQP. Four out of 209 probands (=1.9%) had no detectable sHLA‐RQP. Significant associations of high or low sHLA‐RQP levels to particular HLA‐DR or ‐DQ specificities were not observed. However, plasma derived from HLA‐DR9 positive had the highest and from HLA‐DR8 positive donors the lowest mean sHLA‐RQP values. By comparing HLA identical with two‐haplotype‐different siblings we found no evidence that sHLA‐RQP plasma levels are under genetic control of the HLA complex or closely linked genes. Furthermore, soluble HLA class I plasma concentrations in 100 probands analyzed showed no correlation to those of sHLA‐RQP.
The Canadian Red Cross Plasmapheresis Donor Safety Program: Changes in Plasma Proteins after Long‐Term Plasmapheresis
Authors: Wasi Safia , Santowski Thomas , Murray Shelley A. , Perrault Roger A. , Gill Peter
Published: March 2009

Abstract. The Canadian Red Cross Blood Services have been harvesting plasma from whole blood by plasmapheresis procedure for the last 10 years. To date, we have performed approximately 230,000 procedures. To determine whether this procedure is a health hazard to an individual, a donor safety program was established in 1979 at the National Reference Laboratory. Serum levels of total protein, albumin, and immunoglobulins are monitored at intervals set by the Bureau of Biologics, Health and Welfare Canada. In this communication, we present a 10‐year evaluation of this program. A comparison of the protein concentration distributions between first‐time and long‐term plasmapheresis donors showed no significant differences. Therefore, we have demonstrated that the donors are not at risk as the result of changes in the measured plasma protein levels following plasmapheresis procedure as performed over the last 10 years at The Canadian Red Cross Blood Services.
Extended Storage of Platelets in an Artificial Medium with the Platelet Activation Inhibitors Prostaglandin E1 and Theophylline1
Authors: Bode Arthur P. , Holme Stein , Heaton W. Andrew , Swanson Melvin S.
Published: March 2009

Abstract. The addition of platelet activation inhibitors to the anticoagulant and the replacement of plasma with a fortified electrolyte medium have been shown separately in previous work to improve the storage of platelets during a 2‐week period. In the present study, we have combined these strategies to investigate whether a synergistic improvement could be obtained. A total of 85 concentrates was studied with 300nM prostaglandin E1 (PGE1) and 1.9mM theophylline added to the whole blood, platelet‐rich plasma (PRP), and/or the storage medium during the preparation of platelet concentrates. In vitro markers of platelet aggregation, respiration, and cell integrity were measured over a 20‐day storage period and evaluated in an analysis of variance. We found that a single‐step addition of PGE1 and theophylline to the PRP prior to centrifugation was not sufficient in terms of preventing a rapid fall in pH, rise in pO2, fall in pCO2, loss of hypotonic shock response, and loss of aggregation response, compared to the addition of the inhibitors to the storage medium used to resuspend the platelet pellet. Factorial analysis showed that a reduction in the surface‐to‐volume ratio of the storage container further improved the maintenance of platelet respiration and, for three in vitro markers (hypotonic shock response, released lactic dehydrogenase, and surface glycoprotein Ib levels) displayed an interactive effect with the inhibitors. The addition of protease inhibitors to the formulation of PGE1 and theophylline showed further improvement in several markers. These findings demonstrate the possibility of preserving platelets for 15–20 days with the synergistic effects of activation inhibitors and an electrolyte storage medium fortified with citrate, buffers, and dextrose. In addition, these data suggest that platelet agonists generated in plasma accelerate the in vitro aging process of stored platelets.
Large‐Scale Preparation of a Highly Purified Solvent‐Detergent Treated Factor VIII Concentrate
Authors: Myers Robert , Wickerhauser Milan , Charamella Leigh , Simon Louise , Nummy William , Brodniewicz‐Proba Teresa
Published: March 2009

Abstract. Large‐scale adaptation of a recently reported glycine precipitation method for the production of factor VIII (FVIII) concentrate is described. Scaling up of the method required some modification including the addition of aluminum hydroxide to the glycine buffer to reduce the level of contaminating proteins in the final preparation and the use of centrifugation to replace filtration by glass beads. Furthermore, the resultant product was virus inactivated by incorporation of the organic solvent and detergent technique. At industrial level, the modified method gave a good recovery of FVIII activity (230 IU/l plasma) with high purity (4 IU/mg protein). The final product, after virus inactivation and lyophilization, yielded 185 IU of FVIII activity per liter of starting plasma and was considered to be suitable for clinical evaluation.
Defining the Specificity of Anti‐A/B IgM/G Antibodies with Different Antigens and Lectins1
Authors: Buchs J.‐P. , Nydegger U. E.
Published: March 2009

Abstract. The specificity of anti‐histo blood group A/B antibodies is defined by the antigen used for their production and selection (in the case of mouse monoclonal antibodies, mAbs) or by the antigens present on various intestinal bacteria (for polyclonal human IgM and/or IgG, phAbs). Absorption experiments with red blood cells and free antigen have been used earlier to define specificity; here we use A/B substances of human, animal and synthetic origin as well as known reactivities of mAbs or lectins as tools to further characterize epitope properties of the A/B antigens. The signal response patterns obtained with identical phAbs anti‐A/B IgM/G on three different antigens are superposable. In contrast, commercially available mAb anti‐A/B IgM, when tested on the same antigens, revealed different response curves. The chemical specificity of lectins for distinct monosaccharides in terminal position was exploited to delineate the specificity of mouse ascites anti‐A IgG1 and of phAbs anti‐A/B IgM/G. Helix pomatia lectin inhibited the access of MB9 to porcine A almost completely whereas binding capacities of human anti‐A IgM/G were inhibited by 50 and 62%, respectively. In similar experiments Bandeiraea simplicifolia lectin was seen to inhibit access of anti‐B IgM/G to horse B by 34 and 58%, respectively. No inhibition was seen with lectin from Ulex europaeus or concanavalin A on plates coated with the three different A or B substances. Thin‐layer chromatography (TLC) of antigens revealed one spot for the synthetic trisaccharides, whereas the human and animal blood group substances showed 2 and 3 spots, respectively. Immunostaining experiments of TLC with mAbs revealed 1 spot only per lane, suggesting a high degree of epitope homogeneity.
Elucidation of Apparent Non‐Maternity with DNA Probes Detecting Highly Polymorphic Single Locus Systems
Authors: Fischer G. F. , Pickl W. F. , Faé I. , Pausch V. , Speiser P.
Published: March 2009

Abstract. During paternity testing, we encountered the following constellation in the Jk system: the mother's phenotype was Jk(a‐b+), while the son was typed as Jk(a+b‐). The deduced genotype of the mother would have been Jkb Jkb, and each offspring should then express the Jk(b) antigen. Consequently, non‐maternity would be deduced. Since no material was available for extended family studies or HLA typing, except for the DNA of the propositi, only RFLP analysis could bring clarification in this case. The application of four highly polymorphic single locus probes proved the maternity and hence the existence of a Jk‐Null allele. We conclude that direct testing at the DNA level may help resolving cases where, by conventional parentage testing, conclusive results are unachievable because of putative ‘Null’ alleles.
Pharmacokinetics of an Anti‐Cytomegalovirus Hyperimmunoglobulin after Single Intravenous Administration to Healthy Volunteers
Authors: Ganzinger U. , Martindale J.J. , Gaudera E. , Millendorfer A. , Scriba M. , Bachmayer H.
Published: March 2009

Abstract.By selecting blood donors with high cytomegalovirus (CMV) antibody titres, a plasma pool was obtained which was used to produce an IgG hyperimmunoglobulin by means of pepsin fractionation. After administration of approximately 100 mg/kg body weight to healthy subjects, the time course both of anti‐CMV IgG antibody titres by ELISA and of virus neutralisation (VN) titres was followed for 15 days. Seronegative subjects showed an increase in CMV‐IgG antibodies as well as a significant enhancement of VN. The time course of both titres was non‐uniform. The decline of both titres was biphasic: CMV‐IgG antibodies fell slowly during the first week and remained unchanged thereafter, whereas VN titres decreased markedly faster in the first than in the second week. In seropositive subjects, on the other hand, VN remained unchanged. CMV‐IgG antibodies increased by approximately 3 times, followed by a similar biphasic decline as seen in seronegative subjects. Due to the differences between seronegative and seropositive subjects and to the nonuniform time course, no calculations of the elimination rate were feasible.
Results of Tests with Different Cellular Bioassays in Relation to Severity of RhD Haemolytic Disease
Authors: Mollison P.L.
Published: March 2009

Abstract. Nine different laboratories collaborated in testing sera containing anti‐D using various assays with antibody‐coated red cells, including antibody‐dependent cell‐mediated cytotoxicity, either with monocytes (ADCC(M)) or K‐lymphocytes (ADCC(L)), monocyte‐mediated chemiluminescence and adherence to, and phagocytosis by, peripheral monocytes, U937 cells or cultured macrophages. The samples tested included 14 from 11 mothers of infants with Rh haemolytic disease, 2 monoclonal anti‐Ds and 3 samples containing hyperimmune anti‐D. The results of the bioassays were graded on a scale of 1–3, as were the degrees of severity of haemolytic disease; results were regarded as correct when the grades of the bioassay result and of the degree of severity were the same. Overall, correct results were obtained with the various assays as follows: ADCC(M), 60%; ADCC(L), 57%; chemiluminescence, 50%; adherence and phagocytosis with peripheral monocytes, 41% (but 71% in one laboratory), and with U937 cells or cultured macrophages, 32%. The ADCC(M) assay, compared with the ADCC(L) assay, not only gave better correlations with the severity of haemolytic disease but also gave higher grades of results with hyperimmune sera and with the monoclonal anti‐Ds. Because many of the samples tested were taken after delivery rather than during pregnancy, and because of variations between laboratories, the present results do not conclusively establish the superiority of the ADCC(M) assay in predicting the severity of Rh haemolytic disease.
Fatal Yersinia enterocolitica (Serotype 0:5,27) Sepsis after Blood Transfusion
Authors: Stubbs James R. , Reddy Ramakrishna L. , Elg Steven A. , Perry Elizabeth H. , Adcock Leon L. , McCullough Jeffrey
Published: March 2009

Abstract. Septicemia is a rare complication of blood transfusion. This is probably primarily due to the use of sealed disposable containers for blood collection and the storage of red cell‐containing components at 4°C. However, despite these measures, septicemia due to blood transfusion continues to occur. We report here a fatal case of Yersinia enterocolitica septicemia due to a contaminated unit of red cells which was collected from an apparently healthy, asymptomatic blood donor. The organism grows at cold temperature and multiplies during storage of red blood cell‐containing components. Contaminated components do not show any visible abnormalities. The possibility of transfusion‐transmitted Y. enterocolitica should be considered in patients who have symptoms of sepsis or shock following transfusion.
Distinction of the Glycophorin C Locus from the Diego, Dombrock and Yt Blood Group Loci
Authors: Zelinski T. , Kaita H. , Lewis M. , Coghlan G. , White L. , Cartron J.‐P.
Published: March 2009

Abstract. DNA samples from families informative for the Diego (DI), Dombrock (DO) and Yt (YT) blood group loci were analyzed with a cDNA probe defining a Taq I polymorphism at the glycophorin C locus (GYPC). Recombination between GYPC and DI, DO and YT occurs. Hence GYPC is differentiated from all established blood group system loci.
Recombinant Immunoblot Assay First and Second Generations on 732 Blood Donors Reactive for Antibodies to Hepatitis C Virus by ELISA
Authors: Couroucé Anne‐Marie , Janot Christian ,   The Hepatitis Study Group of the French Society of Blood Transfusion
Published: March 2009

Abstract. A comparison between recombinant immunoblot assay hepatitis C virus (HCV) first generation (RIBA‐1) and second generation (RIBA‐2) was made on 732 blood donors reactive by anti‐HCV ELISA (Ortho Diagnostics System) by the Hepatitis Study Group of the French Society of Blood Transfusion. RIBA‐2 results were correlated with ELISA ratio and ALT levels. The number of both reactive and nonreactive samples was higher with RIBA‐2 than with RIBA‐1, 252 (34%) compared to 224 (31%) for reactive samples, and 404 (55%) compared to 307 (42%) for nonreactive samples. C 22–3 and C 33‐c reactivities were observed in 96 and 91% of the reactive samples, respectively. A total of 76 samples (11%) remained indeterminate by RIBA‐2, 84% of them reacting only on C 100–3 antigen. A clear relationship between RIBA‐2 results and both ELISA ratio and alanine aminotransferase (ALT) levels was demonstrated: 20% of samples with normal ALT level and 10% of samples with low ELISA ratio were reactive when 91% of samples with ALT > 2N and 69% of samples with high ELISA ratio were reactive. The totality of the 57 samples with both ALT > 2N and high ELISA ratio were reactive and 93% of samples with normal ALT level and low ELISA ratio were nonreactive.
Lewis Blood Group Antigens in Salivary Glands and Stratified Epithelium: Lack of Regulation of Lewis Antigen Expression in Ductal and Buccal Mucosal Lining Epithelia1
Authors: Mandel Ulla , Ørntoft Torben F. , Holmes Eric H. , SØrensen Henning , Clausen Henrik , Hakomori Sen‐itiroh , Dabelsteen Erik
Published: March 2009

Abstract. The expression of Lewis antigens is thought to be controlled by the Secretor and Lewis genes. While secretor status is known to regulate the expression of ABH antigens in many tissues, few studies have attempted to correlate Lewis antigen expression on erythrocytes and saliva with that of epithelial tissues. We examined the expression of Lewis a and b and related antigens in human epithelium of minor salivary glands and labial nonkeratinized oral mucosae from 16 individuals by immunohistology using monoclonal antibodies. The expression of these antigens, as detected by monoclonal antibodies (MAbs) used, was correlated with erythrocyte phenotype and saliva secretor status. In acinar cells of glands, Leb antigen was expressed only in secretors, and Lea only in nonsecretors. However, in gland ducts and oral mucosae, Leb was found in both secretors and nonsecretors, as well as in 2 cases of Lea‐b‐, secretors. Thus, antigen expression in acinar cells of minor salivary glands was correlated with the predicted genotypes, whereas inappropriate expression of Leb antigen was found in epithelial cells of gland ducts and oral mucosae. The present data indicate that the Lewis blood group phenotype is regulated differently in duct cells and stratified epithelium than in saliva and erythrocytes.
A Human Anti‐Jkb Monoclonal Antibody
Authors: Lecointre‐Coatmelec M. , Bourel D. , Ferrette J. , Genetet B.
Published: March 2009

Abstract. Peripheral blood mononuclear cells of an immunized patient were transformed with Epstein‐Barr virus and then fused with P3X63Ag8 mouse myeloma cells by polyethylene glycol. After the cloning, a hybridoma cell line secreting specific anti‐Jkb monoclonal antibody was isolated. The antibody was produced in supernatant form and tested for its use as a blood grouping reagent.