bluuebunny/crossref_metadata_2025 huggingface dataset:
Improved Serodiagnosis of Posttransfusion Hepatitis C Virus Infection by a Second‐Generation Immunoassay Based on Multiple Recombinant Antigens
Authors: Wang Jin‐Town , Wang Teh‐Hong , Lin Jaw‐Town , Slieu Jin‐Chuan , Lee Cha‐Ze , Chen Ding‐Shinn
Published: March 2009

Serial serum samples from 35 patients with posttransfusion non‐A, non‐B hepatitis in a prospective study were tested for antibody to hepatitis C virus (HCV) by a multiple recombinant antigen based immunoassay [anti‐HCV (2nd); Abbott], Of them, 23 were positive for anti‐C100, and 27 were positive for HCV RNA by polymerase chain reaction. By anti‐HCV (2nd), 28 patients were positive, and all except 1 of the 28 patients were positive for HCV RNA. A total of 24 patients, who became HCV RNA positive at the acute stage and who were negative for both anti‐C100 and anti‐HCV (2nd) before transfusion, were considered to have posttransfusion hepatitis C. The mean time to seroconversion for anti‐HCV (2nd) was 7.5 or 12.1 weeks after the onset of hepatitis or the date of transfusion, respectively, and was generally 6 weeks earlier than that detected by anti‐C100. However, seroconversion was delayed in 2 hepatitis B surface antigen carriers as compared with anti‐C100. The anti‐C100 assay detected 20 (83%) and the anti‐HCV (2nd) all 24 patients with documented posttransfusion hepatitis C. The second‐generation test is, therefore, better than conventional anti‐C100 for the early diagnosis of HCV infection.
Quantitation of Soluble HLA Class I Antigen in Human Albumin and Immunoglobulin Preparations for Intravenous Use by Solid‐Phase Immunoassay
Authors: Santoso S. , Kiefel V. , Volz H. , Mueller‐Eckhardt C.
Published: March 2009

Soluble HLA class I antigens in human plasma preparations possibly play a role in HLA sensitization and modulation of the immune response. We therefore have determined their concentration in albumin and immunoglobulin preparations from several commercial sources and compared these values to the concentration in normal human sera. For this purpose we used a newly developed solid‐phase enzyme immunoassay employing rabbit anti‐mouse antibody, monomorphic HLA class I monoclonal antibody and a polyclonal enzyme‐linked β2‐microglobulin‐specific antiserum. Soluble HLA antigen concentration in 14 albumin batches from 6 manufacturers and in 16 immunoglobulin batches from 11 manufacturers ranged from 0 to 9.6 and from 0 to 20.9 ng/ml. The concentration in normal human sera 1,328 ± 954 ng/ml (n = 54). We conclude that the concentration of soluble HLA concentration in albumin and immunoglobulin preparations is more than 50 times lower than in normal human serum, but considerable differences exist between products of various manufacturers.
Misleading Results in the Determination of Haemolytic Disease of the Newborn Using Antibody Titration and ADCC in a Woman with Anti‐Lub
Authors: Novotny V. M. J. , Kanhai H. H. H. , Overbeeke M. A. M. , Schlaman‐Nijp A. , Harvey M. S. , Brand A.
Published: March 2009

The Lutheran blood group system consists of several genes of which Lua and Lub are the best characterized. Over 98% of the Western population is Lu (b+), and consequently antibody formation against Lubis rare. Cases reported in the literature suggest that the expression of Lutheran antigens on red cells of the newborn is weak, and Lutheran antibodies are not known to have caused severe haemolytic disease of the newborn. We describe a case of anti‐Lubimmunization during pregnancy. Despite increasing anti‐Lubantibody titre and antibody‐dependent, cell‐mediated cytotoxic activity of the anti‐Lub, two parameters associated with haemolytic disease of the newborn, the baby did not suffer from erythroblastosis.
Evidence that the Low‐Incidence Antigens Termed Wu(700.13) and Hov(700.38) Are Identical
Authors: Moulds Marilyn , Kaita Hiroko , Kornstad L. , Lubenko A.
Published: March 2009

Concordance of reactions of Wu+ and ‘Hov’+ cells with 153 sera containing multiple specificities to low‐incidence antigens indicates that the ‘two’ antigens are identical. This conclusion is confirmed by absorption and elution studies.
Comparison of Five Different Filters for the Removal of Leukocytes from Red Cell Concentrates
Authors: Pietersz R.N.I. , Reesink H. W. , Dekker W. J. A. , Steneker I. , Biewenga J. , Al E. J. M. , Huisman J. G.
Published: March 2009

The leukocyte depletion capacity and performance of 5 filters designed for filtration of red cell concentrates (RCC) were compared by counting leukocytes, measuring red cell volumes and by histological examination of the filters after use. To eliminate interdonor differences, 5 buffy‐coat‐poor RCC were pooled (in each of 10 experiments) and subsequently split up into the original bags. The RCC were passed over the Cellselect filter, a column filled with cellulose acetate, and over flat‐bed polyester filters: the Cellselect Optima, the Pall RC 50, the Leukostop and the Sepacell R‐500.

The filtration was shortest with the Pall RC 50 (p < 0.001 compared to the other 4 filters). Leukocyte removal was most effective with the cellulose acetate filter (p < 0.01 compared to the other 4 filters) followed by the Cellselect Optima polyester filter (p < 0.02 compared to the remaining 3 filters). Residual leukocytes did not exceed 50 × 106 for any brand of filter. Red cell recovery was similar for all 5 filters with mean values from 86.1 to 89.2%.

The leukocyte numbers, counted in Türk's solution or in propidium iodide, gave comparable values in hemocytometers applying light microscopy or fluorescent microscopy, respectively.

Histological examination showed that lymphocytes were mainly removed by trapping, whereas granulocytes showed a variable pattern: adhesion in presence of platelets or trapping.
Feasibility of a Predeposit Autologous Blood Donation Program in Colorectal Cancer Patients: Results from a Randomized Clinical Study
Authors: van Papendrecht M. A. W. Hoynck , Hop W. , Langenhorst B. L. A. M. , Kothe F. C. , Marquet R. L. , Jeekel J.
Published: March 2009

The hematologic and transfusion data of a multicenter randomized trial investigating the effect of blood transfusions on the 5‐year survival were used to study the feasibility of an autologous blood donation program in colorectal cancer patients. Three hundred and ten patients were randomized for autologous blood transfusions (predeposition of 2 units) or homologous blood transfusions, and transfusion rules were standardized.

The Hb level in the patients who donated blood decreased by 20.1 ± 1.3 g/1 (mean ± SEM) preoperatively and 4.5 ± 1.8 g/1 postoperatively, and in controls 3.7 ± 1.1 g/1 and 16.5 ± 1.9 g/1 (significantly different between the two groups, both pre‐ and postoperatively: p < 0.01). Because blood loss and number of transfusions were similar in both groups, this indicated that either preoperative or postoperative erythropoiesis is stronger in patients who had donated blood. Twenty‐three percent of the autologous patients and 61% of the homologous patients were exposed to homologous blood. The effectiveness of the procedure differed per tumor localization. In patients with a right‐sided colon carcinoma, 22% of the control patients needed homologous blood, compared to 10% of the autologous patients. In patients with other colon carcinomas, this was 52 and 16%, respectively, and in patients with a rectal carcinoma 85 and 41%.

We conclude that predeposition of 2 units of blood for colorectal cancer surgery is feasible and useful to prevent homologous blood usage in a significant number of patients with left colon carcinoma or rectal carcinoma.
A Case of ‘Auto‐Immune Thrombocytopenic Purpura’ Serologically Similar to Post‐Transfusion Purpura
Authors: Taaning E. , Killmann† S.A.
Published: March 2009

The report describes a HPA‐1a (Zwa)‐negative woman with thrombocytopenia and antibodies in serum and eluate from autologous platelets with an operational anti‐HPA‐1a specificity. The results of the serological investigations were similar to the findings in most patients suffering from post‐transfusion purpura. However, the present patient had no history of blood transfusion prior to the unset of purpura and the thrombocytopenia had persisted 6 months before splenectomy.
Analysis of the Gene Polymorphism of ABO Blood Group Specific Transferases Helps Diagnosis of Acquired B Status
Authors: Fischer G. F. , Faé Ingrid , Dub Elisabeth , Pickl W. F.
Published: March 2009

Blood group typing of an aged patient suffering from ileus provided evidence for an acquired B. As a parameter independent of cell membrane molecules or secreted blood group substances, the nucleotide sequence polymorphism of A and B transferases was investigated. Restriction fragment length polymorphism of DNA chains amplified in a polymerase chain reaction from the coding region of the glycosyltransferase indicated that no gene for B transferase was present in the patient's genome. We conclude that the assessment of polymorphism of ABO blood group transferase can be used as a marker independent of blood group molecules for confirming a suspected acquired B.
Influence of the Cryoprotective Agents Glycerol and Hydroxyethyl Starch on Red Blood Cell ATP and 2,3‐Diphosphoglyceric Acid Levels
Authors: Rittmeyer I. C. , Nydegger U. E.
Published: March 2009

Because hydroxyethyl starch (HAES) is used for volume replacement therapy and as a cryoprotectant for frozen red blood cells (RBCs), this compound, in contrast to glycerol, does not require labor‐intensive removal from thawed cells prior to transfusion. We here report the effect of both glycerol and HAES on the RBC organic phosphates ATP and 2,3‐diphosphoglyceric acid (2,3‐DPG). The CPD‐A1‐stabilized RBCs of 20 healthy donors (3 females, 17 males) were separately frozen in either 40% glycerol or 6% HAES, of molecular weight 200,000. ATP and 2,3‐DPG concentrations were determined in CPD‐A1 RBCs before addition of cryoprotectant and in cryopreserved thawed RBCs after 24 h storage at ‐80°C (glycerol) and ‐196°C (HAES). It appears that HAES, but not glycerol, significantly reduces ATP concentrations whereas both lead to a reduction of 2,3‐DPG concentrations; this reduction was more pronounced with glycerol than with HAES. Experiments with the blood of 6 donors demonstrated that HAES affects autohemolysis by 16%, in contrast to glycerol, after which cryoprotectant autohemolysis was affected by 3.1% only. RBC recoveries were comparable using glycerol or HAES as cryoprotectants. A distinct pattern of reduction of 2,3‐DPG levels by glycerol and less by HAES, and of ATP levels by HAES but not by glycerol, emerges. Our findings may be of importance if HAES is to be introduced as a convenient cryoprotectant.
Point Mutation in the Glycophorin C Gene Results in the Expression of the Blood Group Antigen Dha
Authors: King M.J. , Avent N.D. , Mallinson G. , Reid M.E.
Published: March 2009

The blood group Duch (Dha) antigen is located on glycophorin C (GPC). Total RNA prepared from the reticulocyte fraction of two Dh(a+) individuals were used in the synthesis of first‐strand cDNA. The first‐strand cDNA served as templates for the amplification of GPC‐related DNA by polymerase chain reaction (PCR). The expected PCR product consisted of 412 base pairs. On sequencing the PCR‐amplified DNA, a base change (cytosine→thymidine) at nucleotide 40 of the GPC cDNA was detected. Thus, the variant GPC (GPC. Dha) on Dh(a+) red cells has a substitution of leucine by phenylalanine at amino acid residue 14.
Transfusion Management of Patients with IgA Deficiency and Anti‐IgA during Liver Transplantation
Authors: Davenport R.D. , Burnie K.L. , Barr R.M.
Published: March 2009

Severe anaphylactic or allergic reactions may occur during blood transfusion to patients who are IgA‐deficient and have anti‐IgA in their blood, particularly those with class‐specific antibodies. These patients are a particular challenge to the hospital transfusion service when large volumes of blood components are required for transfusion support, as in liver transplantation. We have successfully provided blood components for 3 such patients undergoing liver transplantation. Red cells were washed manually or by automated technique. Platelets were washed manually. All plasma was from IgA‐deficient donors. One patient's entire plasma requirements were supplied by autologous plasmapheresis. Serial determinations of IgA levels and anti‐IgA titers in 1 patient demonstrated an abrupt fall in anti‐IgA with the appearance of barely detectable amounts of IgA during the surgery. IgA‐containing plasma cells were demonstrated in the biopsies of liver homografts of 2 patients following transplantation. IgA deficiency with anti‐IgA can be successfully managed during liver transplantation with advance planning.
The Quantification of Fetomaternal Hemorrhage by an Enzyme‐Linked Antibody Test with Glutaraldehyde Fixation
Authors: Greenwalt T.J. , Dumaswala U.J. , Domino M.M.
Published: March 2009

A modified enzyme‐linked antiglobulin test (ELAT) for quantifying D‐posi‐tive red blood cells in the circulation of D‐negative pregnant women is described. The antibody‐antigen bond was stabilized and the problem of hemolysis eliminated by the use of 0.05% glutaraldehyde. The r value for the standardization curves for measuring cord RBC in mixtures was 0.98; coefficient of variation 5.8%. Reproducible quantitation of 0.125% D‐positive cord RBC mixed with D‐negative RBC was demonstrated. In comparative studies of artificial mixtures, Kleihauer‐Betke results for more than 1% cord RBC were 50–100% greater than the true values. In several examples of large fetomaternal hemorrhages, the ELAT results matched the clinical data more closely than the Kleihauer‐Betke based estimates. 2 ml of fetal RBC in a 1,600‐ml red cell mass can be quantified using the modified ELAT.
A Decade of Rare Donor Services in the United States
Authors: Mallory D. , Malamut D. , Sandler S. G.
Published: March 2009

Between 1981 and 1990, the American Red Cross Rare Donor Registry supplied 9,872 units of red cell components with rare phenotypes to blood centers in the United States and abroad. Approximately 51% were from donors with high‐frequency antigen‐negative phenotypes and 49% were from donors with multiple antigen‐negative phenotypes. Since 1989, the disease category requiring the largest number of units has been sickle cell disease. Strategies to ensure that the Registry will have adequate resources to meet future requirements include testing selected donors for rare phenotypes and blood conservation programs, such as intraoperative salvage and the treatment of anemia of chronic renal failure with recombinant erythropoietin.
Serological Screening, Using Three Different Test Systems of Platelet‐Transfused Patients with Hematologic‐Oncologic Disorders
Authors: Wernet Dorothee , Schnaidt Martina , Mayer Gerhard , Northoff Hinnak
Published: March 2009

Sera of hematologic‐oncologic patients were tested regularly after platelet transfusions in three test systems: lymphoeytotoxicity test, platelet adhesion immunofluorescence test, and — only selected sera — in the monoclonal antibody‐specific immobilization of platelet antigen test. Of 388 patients 53 (14%) had HLA antibodies 5 of these in combination with platelet‐specific alloantibodies. Lymphocyte‐restricted (non‐HLA) reactions were observed in 20 patients, the majority of which was attributed to lymphocyte‐specific auto‐ or alloantibodies. Sera of 27 patients showed platelet‐specific reactions, usually cold‐reacting autoantibodies which have no effect in vivo.
Heparin‐Associated Thrombocytopenia: No Association of Immune Response with HLA
Authors: Greinacher A. , Mueller‐Eckhardt G.
Published: March 2009

Heparin‐associated thrombocytopenia (HAT) is a severe adverse reaction of heparin therapy. Patients with the immunologic type of HAT are at risk of developing arterial and venous thromboembolic complications caused by an antibody which activates platelets in the presence of heparin. Yet, there are no means to identify patients at risk of developing HAT before heparin administration. We investigated the frequency of HLA class‐I and class‐II antigens in 47 patients with the immunologic type of HAT verified by a positive two‐point heparin‐induced platelet activation assay. Compared to a control group of 629 healthy individuals HLA antigens B8 and DR3 were less frequent, whereas DR4 and DR53 had a higher frequency. However, none of these antigens proved to be statistically significant. This is another example of an immunologically mediated disease showing no discernible association with the HLA system.
Effectiveness of Four Intravenous IgG Preparations in Prevention of Phagocytosis of Anti‐Rh‐(D)‐Coated Erythrocytes by Mononuclear Phagocytes
Authors: Sheth Kirtikant V. , Al‐Sedairy Sultan , Lee Jennifer
Published: March 2009

Four commercial intravenous immunoglobulin G (IGIV) preparations, Sandoglobulin (Sandoz), Intraglobin (Biotest), Gammonativ (Kabivitrum) and Gammaogard (Kabivitrum) showed percent inhibition of Fc receptor‐mediated phagocytosis of anti‐Rh(D)‐coated erythrocytes as follows: 45±14 SD, 41±23 SD, 35±14 SD, 25±13 SD, respectively. Pre‐incubation of Rh(D)‐positive and antibody‐coated Rh(D)‐positive erythrocytes with IGIV did not lead to any inhibitory effects on phagocytosis. However, preincubation of mononuclear phagocytes with IGIV gave marked inhibition of phagocytosis of antibody‐coated erythrocytes. In summary, all four IGIV preparations showed Fc receptor‐mediated inhibition of phagocytosis of anti‐Rh(D)‐coated erythrocytes by mononuclear phagocytes.
Preparation of Intermediate‐Purity Factor VIII Concentrate by Direct Gel Filtration of Cryoprecipitate
Authors: Teh L. C.
Published: March 2009

We report a new method to produce a solvent/detergent‐treated and severe dry heat‐treated factor VIII (FVIII) concentrate (3‐6IU FVIII:C/mg protein). This method, which uses a single purification step after cryoprecipitation, is suitable for scale‐up to production levels. FVIII was obtained from solvent/detergent‐treated cryoprecipitate by a single gel filtration step using Sephacryl S‐400HR. The freeze‐dried product was stable to heating at 80°C for 72 h. The yield of the solvent/detergent and severe dry heat‐treated product was 230 IU FVIII: C/kg plasma. The reconstituted product gave a 10% loss in FVIII: C activity after heating at 37°C for 6 h. The feasibility of this method suggests that gel filtration using S‐400HR can be used solely or as part of a purification process for the preparation of high‐purity FVIII concentrates.
HLA DNA Preorgan Retrieval Donor Typing in Renal Transplantation
Authors: Gläser Regine , Eiermann Thomas H. , Fricke Lutz , Kirchner Holger , Bein Gregor
Published: March 2009

For the first time, a DNA technique was used in clinical practice for preorgan donor retrieval typing in renal transplantation. The method is based on the polymerase chain reaction with nested sequence‐specific primer pairs (nested PCR‐SSP) for all serological HLA‐DR specificities of DR1‐DR18. A panel of reference‐typed individuals (n = 101) was investigated in a blind quality control study and the results revealed a sensitivity of 99.5% and a specificity of 100%. Semi‐automation of PCR‐SSP provided rapid, accurate and reliable typing results demonstrating that this DNA test is most suitable for replacing the error‐prone HLA‐DR serological technique. Twenty‐seven consecutive cadaveric organ donors have been successfully typed so far. The results were available within less than 3 h after blood sampling and in all cases prior to the retrieval of organs. In conclusion, the introduction of this accurate DNA typing in HLA matching programs may significantly improve the graft survival rate in renal transplantation.
Orosomucoid System: 17 Additional Orosomucoid Variants and Proposal for a New Nomenclature
Authors: Yuasa I. , Weidinger S. , Umetsu K. , Suenaga K. , Ishimoto G. , Eap B.C. , Duche J.‐C. , Baumann P.
Published: March 2009

There are two forms of orosomucoid (ORM) in the sera of most individuals. They are encoded by two separate but closely linked loci, ORM1 and ORM2. A number of variants have been identified in various populations. Duplication and nonexpression are also observed in some populations. Thus, the ORM system is very complicated and its nomenclature is very confusing. In order to propose a new nomenclature, ORM variants detected by several laboratories have been compared and characterized by isoelectric focusing (IEF) followed by immunoprinting. A total of 57 different alleles including 17 new ones were identified. The 27 alleles were assigned to the ORM1 locus, and the others to the ORM2 locus. The designations ORM*F1, ORMI*F2, ORMI*S and ORM2*M were adopted for the four common alleles instead of ORM1*1, ORM1*3, ORM1*2 and ORM2*1 (ORM2*A), respectively. The variants were designated alphanumerically according to their relative mobilities after IEF in a pH gradient of 4.5‐5.4 with Triton X‐100 and glycerol. For the duplicated genes a prefix is added to a combined name of two alleles, e.g. ORM1*dB9S. Silent alleles were named ORM1*Q0 and ORM2*Q0 conventionally. In addition, the effects of diseases to ORM band patterns after IEF are also discussed.
Posttransfusional Hemolysis in Recipients of Glucose‐6‐Phoiphate Dehydrogenase‐Deficient Erythrocytes
Authors: Shalev Oded , Manny Noga , Sharon Raphael
Published: March 2009

To test the hypothesis that transfusion of blood donated by individuals with glucose‐6‐phosphate dehydrogenease (G6PD) deficiency may result in a hemolytic reaction, we conducted a prospective longitudinal study in which 10 patients transfused with 1 unit of G6PD‐deficient and 1 unit of normal red blood cells (RBC) were compared with 10 patients transfused with 2 units of age‐matched normal RBC. We found that 24 h after transfusion serum bilirubin (μmol/1) in the recipients of G6PD‐deficient RBC was significantly higher than in the recipients of normal RBC (36±14 vs. 18±5, respectively, p>0.004). A parallel increase was found in the serum lactate dehydrogenase (LDH; IU/1) between the two groups (378±151 vs. 264±56, p<0.001). The difference in serum bilirubin(26±10 vs. 15±5, p<0.03) was still noted 48 h after transfusion, with only a marginal difference (p<0.08) in LDH. We conclude that an immediate posttransfusional hemolytic reaction can occur in recipients of G6PD‐deficient RBC and therefore suggest that the differential diagnosis of posttransfusional hemolysis, particularly in populations where G6PD deficiency is prevalent, includes transfusion of erythrocytes from G6PD‐deficient donors.
A Flow Cytometric Approach to Quantitative Estimation of Platelet Surface Immunoglobulin G
Authors: Christopoulos C.G. , Kelsey H.C. , Machin S.J.
Published: March 2009

Flow cytometry (FC) was used to estimate platelet‐surface IgG (PSIgG) by quantifying the fluorescence of platelets incubated with a fluorescein isothio‐cyanate (FLTC)‐labelled polyclonal goat anti‐human IgG antibody or FITC‐labelled non‐immune goat IgG. Results were expressed as relative fluorescence intensity (RFI) defined as the ratio of specific fluorescence (mean fluorescence of platelets incubated with the FITC anti‐IgG) over non‐specific fluorescence (mean fluorescence of platelets incubated with FITC non‐immune goat IgG). A normal range was formed by analysing platelets from 71 healthy subjects. Platelets from 16 patients with a firm clinical diagnosis of immune‐mediated thrombocytopenia had a mean RFI significantly higher (p <0.001) than the controls, whereas platelets from 9 patients thought to have non‐immune thrombocytopenia had an RFI not significantly different from the normal controls. From a prospectively studied group of 62 patients with no clinically obvious cause for their thrombocytopenia or impaired platelet function 35.5% had raised PSIgG. In order to express the results as number of IgG molecules per platelet, reference curves were created by using FC to nieasurc PSIgG of platelets coated with known amounts of a chimeric IgG (human IgG with murine hypervariable region) monoclonal antibody to the glycoprotein IIb‐IIIa complex. Normal platelets had an average 1,463 (SD = 927) molecules of PSIgG. In patients with immune‐mediated thrombocytopenia the levels ranged from 690 to 32,328 (mean 11,535) molecules per platelet.

Flow‐cytometric PSIgG estimation was sensitive, fast and easy to perform and therefore suitable for both research and clinical service purposes.
Molecular Genetic Analysis of the ABO Blood Group System: 1. Weak Subgroups: A3 and B3 Alleles
Authors: Yamamoto Fumi‐ichiro , McNeil Patricia D. , Yamamoto Miyako , Hakomori Sen‐itiroh , Harris Teresa , Judd W. John , Davenport Robertson D.
Published: March 2009

We have determined the nucleotide sequences of the coding region in the last two coding exom of ABO genes (which occupy 91% of the soluble form of A1 transferase) from 7 individuals with weak subgroup phenotypes. Four of the individuals had an A3 phenotype and 3 individuals had a B3 phenotype. We determined the nucleotide sequences based on PCR followed by subcloning and DNA sequencing of the amplified fragments. Two cases of the A3 allele and 1 case of the B3 allele were found to contain a single‐base substitution which resulted in an amino acid substitution. However, no other cases of A3 and B3 alleles were found to contain differences in this region. This finding demonstrates for the first time heterogeneity among these weak subgroups at the nucleotide level.
Ultraviolet Irradiation Inhibits Killer‐Target Cell Interaction
Authors: Kobata Tetsuji , Ikeda Hisami , Ohnishi Yoshiko , Urushibara Noriko , Nakata Sayuri Otsuka , Takahashi Tsuneo A. , Sekiguchi Sadayoshi
Published: March 2009

The effects of ultraviolet (UV) irradiation on cell‐mediated cytolysis were examined in order to clarify the inhibitory mechanisms of allosensitization by UV irradiation. UV‐B‐irradiated target cells (Sa; an Epstein‐Barr virus‐transformed B cell line) exhibited more resistance against alloreactive cytotoxic T lymphocytes (CTL) than mitomycin C (MMC)‐treated target cells. In the conjugate formation assay, UV‐B‐irradiated target cells showed a considerably lower binding to alloreactive CTL than MMC‐treated target cells.

UV‐B irradiation induced a reduction of HLA‐class I, ‐DR, CD54 (ICAM‐1) and CD58 (LFA‐3) expression on target cells. However, it does not seem to contribute to the inhibition of cell adhesion induced by UV‐B irradiation because a similar reduction of cell surface antigens was observed in MMC‐treated target cells. Number of cells capped with anti‐HLA‐class I, ‐DR, CD54 or CD58 monoclonal antibody were markedly reduced by UV‐B irradiation compared to that by MMC treatment. These findings suggest the possibility that the inhibition of cell adhesion between UV‐B‐irradiated Sa target cells and alloreactive CTL is due to the impaired mobility of cell surface antigens which will affect the early process of cell‐mediated cytolysis.
Immunofixation Electrophoresis: A Technique for the Study of Protein Polymorphism
Authors: Alper C. A. , Johnson A. M.
Published: March 2009

A technique is described which allows direct visualization of individual proteins in mixtures by specific antiserum after electrophoresis. By minimizing diffusion it permits rapid, direct, and clear detection of genetic polymorphism and ‘conversion’ of proteins in the complement and coagulation systems.
Frozen Blood and Transfusion‐Transmitted Hepatitis C Virus
Authors: Norol F. , Bachir D. , Bernaudin F. , Beaujean F. , Desforges L. , Duedari N. , Galacteros F.
Published: March 2009

The purpose of the present study was to evaluate the impact of frozen red cell transfusion on the transmission of hepatitis C virus (HCV) before the introduction of blood donor screening. Anti‐HCV antibodies were detected in 59 patients with sickle‐cell disease who required chronic transfusions and had exclusively received frozen red blood cells (RBC). The files were reviewed for clinical signs of chronic hepatitis C. Anti‐HCV antibodies were detected in 2 adult patients; both also had clinical evidence of HCV infection. No other patient showed signs of acute or chronic HCV hepatitis. In a control group of 28 patients who had received nonfrozen RBC transfusions, the prevalence of anti‐HCV antibodies was 25%. So, our study seems to indicate that the use of frozen RBC had reduced the risk of HCV contamination.
Production of Stable Human‐Mouse Hybridomas Secreting Monoclonal Antibodies against Rh D and c Antigens
Authors: Rapaille André , François‐Gérard Christiane , Donnay Danielle , Sondag‐Thull Danièle
Published: March 2009

Peripheral blood lymphocytes from donors immunized against Rh antigens were fused with mouse myelomas and heteromyelomas in order to obtain human‐mouse hybridomas secreting antibodies specific for these antigens. Three cell lines secreting anti‐D IgG and two secreting anti‐c IgM were stabilized and produced immunoglobulins for several months. These human monoclonal antibodies were evaluated as reagents for Rh phenotyping. Their complementary activity towards weak D and partial D antigens is examined.
Positive Antiglobulin Test in Association with Sulindac: Involvement of the Rh Factor
Authors: DeCoteau J. , Reis M. D. , Pinkerton P. H. , Ward J. , Coovadia A.S.
Published: March 2009

We investigated the development of a positive direct antiglobulin test associated with the use of the nonsteroid anti‐inflammatory drug sulindac. The drug was shown to be the cause of the positive direct antiglobulin test using red cells treated in vitro with solutions of native sulindac and its two major metabolites. Serum from the patient contained sulindac‐dependent red cell antibodies which could not be demonstrated when red cells having the Rh phenotype D– were employed in the test procedure. An eluate prepared from the patients' red cells reacted against untreated red cells having common Rh phenotypes, but not against target red cells with the Rh phenotypes D– or Rh null. The eluate showed stronger reactivity against the cells having common Rh phenotypes when they were treated with solutions of a metabolite of sulindac, but failed to react against treated red cells having the Rh phenotype D– or Rh null. The results of our investigations point to an interaction between sulindac and/or its metabolites and Rh structures on the red cell membrane as the initial step in the production of drug dependent and autoantibodies leading to the positive direct antiglobulin test.
Effect of Monosaccharides during Severe Dry Heat Treatment of Coagulation Factor VIII Concentrates
Authors: Knevelman A. , De Wit H.J.C. , Griffin B. , Hart H. , McIntosh R.V.
Published: March 2009

During product development of a factor VIII concentrate (Dutch blood banks) the conversion from unsterilized to autoclaved freeze‐drying buffer caused impaired product characteristics after severe dry heat treatment (80°C for 72 h). Analysis of the freeze‐drying buffers showed the presence of fructose and glucose in heated buffers, resulting from hydrolysis of sucrose. The detrimental effect of glucose and fructose on solubility, yield of factor VIII and color of the heat‐treated product was confirmed by freeze‐drying and heating products spiked with increasing levels of these monosaccharides. The effect of the use of freeze‐drying buffers autoclaved with and without sucrose was examined in two other factor VIII concentrates, S(8) and Z8 (Protein Fractionation Centre, Edinburgh, UK). If sucrose was present during autoclaving of the buffer, a slightly lower yield over freeze‐drying and 80°C heat treatment was observed.

Since glucose is present as a substrate in the medium for the host cells during cultivation of viruses, its potential effect on the 80°C heated product (Dutch blood banks) was examined during the validation study of the inactivation of HIV‐1 and Sindbis. The cultivation cycles for the virus inolcula were simulated and residual glucose levels measured. In the supernatant medium of the host cell culture used for the propagation of Sindbis no residual glucose was found. Glucose however was found in the supernatant medium of the host cell culture for propagation of HIV‐1, and therefore small molecular weight substances were removed from the actual HIV‐1 inoculum by ultrafiltration. The study of the inactivation of both viruses during freeze‐drying and 80°C heat treatment was subsequently carried out without complications.
Serological Characteristics of Partial D Antigen Category VI in 8 Unrelated Blood Donors
Authors: van Rhenen D.J. , Thijssen P.M.H.J. , Overbeeke M.A.M.
Published: March 2009

Classification of subjects with a partial D antigen is traditionally performed with immune anti‐D sera. The development of monoclonal antibodies enables a fine analysis to be made of the specificity of the epitopes that are present or missing in these cases. A systematic search in a Caucasian donor population of 17,500 revealed 8 unrelated male individuals (frequency 0.05%) with a red cell phenotype characteristic of partial D category VI, but without anti‐D in their serum. The relation to the ‘classic’ partial D category VI was investigated and is discussed, as is the observed serological heterogeneity of the partial D category VI group. Clinical consequences for the prevention of immunization of these subjects are mentioned.
Complement Killing of Yersinia enterocolitica and Retention of the Bacteria by Leucocyte Removal Filters
Authors: Gong J. , Rawal B. D. , Högman C. F. , Vyas G. N. , Nilsson B. , Gustafsson I.
Published: March 2009

We report studies on the complement sensitivity of four strains of Yersinia enterocolitica, serotypes O:3, O:9, O:5,27, and O:20, isolated from blood units involved in transfusion fatalities. Complement in fresh CPD plasma killed Y. enterocolitica within 4 h at 22°C in 100% of the experiments. The bactericidal action was serotype and complement activation pathway dependent. Both classic and alternate pathways seemed to be active, but the latter to a lesser degree. When the classic pathway was blocked by chelation of Ca2+ no complete killing was obtained. Complement did not enhance or condition Yersinia for leucocyte filter retention. Direct removal of Yersinia by filtration was also related to serotype; all strains were reduced by filtration in heat‐inactivated plasma, and all except serotype O:5,27 were reduced in Ca2+ ‐chelated plasma. Our findings may explain why plasma products and platelet concentrates are rarely involved in Yersinia sepsis related to transfusion.
Hepatitis C Virus Antibodies and Virus Replication in Asymptomatic Blood Donors
Authors: Yuki Nobukazu , Hayashi Norio , Hagiwara Hideki , Naito Masafumi , Ohkawa Kazuyoshi , Kasahara Akinori , Fusamoto Hideyuki , Ohtani Satoshi , Okubo Yasuto , Kamada Takenobu
Published: March 2009

We assessed hepatitis C virus (HCV) infection in 99 asymptomatic blood donors positive using a first‐generation HCV antibody assay. When tested with second‐generation assays, 86 (87%) donors were reactive (group 1), 2 (2%) were indeterminate (group 2), and 11 (11%) were non‐reactive (group 3). Viraemia was revealed by polymerase chain reaction in all group 1 cases. The 2 group 2 cases and 6 (55%) group 3 cases were also viraemic. Viraemia was confirmed by a branched DNA assay in the 2 group 2 cases and 4 (36%) group 3 cases. Serum HCV RNA levels were further studied using a competitive reverse transcription‐polymerase chain reaction assay. All cases in groups 2 and 3 were low viraemic (range 104–105.5copies/ml) compared with the 9 group 1 cases examined (range 107–109copies/ml). No correlation was evident between viraemic levels and antibody cut‐off index in the first‐generation assay. These findings indicate the possibility that low levels of viraemia can occur in individuals non‐reactive in second‐generation HCV antibody assays.
Characteristics of Anti‐A and Anti‐B in Black Zimbabweans
Authors: Adewuyi James O. , Gwanzura Christine , Mvere David
Published: March 2009

Sera from young, black, group 0 Zimbabwean blood donors were screened for anti‐A and anti‐B haemolysins. Nearly one fifth of the sera were found to be strongly haemolytic for either A or B cells or both. Some of the sera were titrated for agglutination in saline before and after treatment with dithiothreitol. Serum dilutions beyond the endpoint of agglutination were further tested by the indirect antiglobulin technique using specific anti‐IgM and anti‐IgG sera. More than 60% of the strongly lytic sera had high titres of IgG (≥ 64). The IgM and IgG concentrations both of anti‐A and anti‐B were correlated and these levels were in turn correlated with haemolytic activity.
Circulating Stem Cell Collection in Lymphoma and Myeloma after Mobilization with Cyclophosphamide and Granulocyte Colony‐Stimulating Factor for Autologous Transplantation
Authors: Cancelas J.A. , Hernández‐Jodra M. , Zamora C. , Perez‐Oteyza J. , Brieva J.A. , Roldan E. , Navas G. , Garcia‐Laraña J. , Lopez J. , Odriozola J.
Published: March 2009

We report the results of 72 leukapheresis procedures performed for autologous peripheral blood stem cell collection in 18 patients with lymphoma and myeloma, after combined mobilization with cyclophosphamide and granulocyte colony‐stimulating factor (G‐CSF). The numbers of mononuclear cells (MNCs), CD34+ cells and granulocyte‐macrophage colony‐forming units (CFU‐GM) either in the peripheral circulation (preleukapheresis sample) or in the product obtained from leukapheresis (leukapheresis sample) were evaluated. A highly superior proportion of CD34+ cells (14‐fold) and CFU‐GM (5‐fold) resulted from the mobilization therapy. CFU‐GM and CD34+ cells were highly enriched with respect to all MNCs (relative recoveries: 2.13, range 0.3–41, and 1.08, range 0.2–8.5, respectively) due to an additional mobilization effect by the leukapheresis procedure. Also, a relatively strong linear correlation between the three different parameters was found in the leukapheresis product (CD34+:CFU‐GM, r = 0.81; MNCs:CD34, r = 0.69; MNCs:CFU‐GM, r = 0.75; CFU‐GM:CD34+, and MNCs, r = 0.85). Our data suggest that the number of MNCs and CD34+ cells obtained after combined mobilization with cyclophosphamide and G‐CSF can be used as predictor of the number of granulomonocytic progenitors.
Serotype Switching in a Partially Deleted RHD Gene
Authors: Blunt Tracy , Daniels Geoff , Carritt Ben
Published: March 2009

We have studied the RH genes in donors with the RhD‐negative haplotype dCes. In contrast to the usual arrangement of genes in RhD‐negative individuals, where the lack of antigen expression is due to deletion of the entire RHD gene, we find that the dCeshaplotype includes an RHD gene with an internal deletion. Moreover, there appear to be no 5′ sequences characteristic of RHC suggesting that the RhC antigen may be encoded by a truncated RHD or a recombinant RHD/CE gene in these dCes/dce genomes.
Effect of Agitation on the Quality of Platelet Concentrates
Authors: Mitchell S.G. , Hawker R.J. , Turner V.S. , Hesslewood S.R. , Harding L.K.
Published: March 2009

Platelet concentrates (PCs) were stored for 4 days at 22°C in 400 ml second‐generation (PL1240) platelet packs with either constant agitation, manual mixing once every 24 h or without agitation at any time. After 4 days storage, in vivo recovery, survival and biodistribution were determined following indium‐111 labelling of platelets and infusion into autologous volunteers. In vitro assays of platelet function and biochemistry were likewise carried out after 4 days storage. The PCs stored without agitation had significantly lower in vivo recoveries, pH and aggregation responses to ionophore A23187 and a combination of collagen and epinephrine and significantly higher β‐thromboglobulin and indium‐111 release than the agitated PCs. The manually mixed PCs were not significantly different from the constantly agitated PCs. PCs mixed simply once every 24 h remained viable with active oxidative phosphorylation and a pH above 6.74 in all but 1 case indicating that PCs stored at 22°C for up to 4 days with only intermittent mixing are satisfactory for transfusion. A change from constant agitation would reduce capital costs in mixing equipment and simplify the transport of PCs from the transfusion centre to small hospital blood banks.
Direct Capture of Plasma Factor VIII:C by Ion Exchange Chromatography
Authors: Teh L.‐C. , Froger M.
Published: March 2009

An ion exchange medium, DEAE‐Fractogel 650M, can be used to capture factor VIII (FVIII) directly from plasma. Previous reports have focussed on the use of this medium to capture FVIII from cryoprecipitate. In this report, citrate phosphate dextrose plasma was batch‐stirred with DEAE‐Sephadex A50, filtered, diluted, loaded onto a column packed with DEAE‐Fractogel TSK 650M, and chromatographed. Most of the unwanted proteins flowed through the gel unadsorbed. Bound FVIII was eluted by increasing the ionic strength of the buffer. A citrate‐based buffer gave an overall FVIII:C yield of 670 IU/kg plasma with a specific activity of 0.68 IU FVIII:C/mg protein. This process gave a higher yield of FVIII:C but significantly more prothrombin complex factors than cryoprecipitation. This method is a promising alternative to cryoprecipitation of FVIII.
The Effect of Previous Blood Transfusion on Lymphocyte Subsets and Natural Killer Cell Function in Patients with Colorectal Cancer
Authors: Mathiesen O. , Lund L. , Brodthagen U. , Gandrup P. , Grunnet N. , Balslev I. , Jersildy C.
Published: March 2009

To elucidate the possible influence of previous blood transfusion on immune functions, the transfusion history of 153 patients admitted to hospital for elective colorectal surgery was correlated with lymphocyte subsets and natural killer (NK) cell function. The subsets determined were CD2, CD3, CD4, CD8, CD16, CD20, CD56, CD57 and HLA‐DR‐positive. The NK cell function was determined by measuring the killing capacity against cFDA‐labelled K562 target cells monitored via a flow‐cytometer. We found that 42 patients (27%) had been transfused before surgery, of these 13 had been transfused less than 30 days before surgery and 29 (19%) transfused more than 30 days before (median 10 years, range 0.1–37 years). In transfused patients, we found a significantly reduced number of B lymphocytes (CD20; p = 0.01), a reduction in HLA‐DR‐positive cells (p = 0.02) and a just significant reduction of NK cell function in transfused compared to nontransfused patients. The reduction in NK cell function is marginal and the NK cell function is within normal range, and probably without clinical significance. Reduction in NK cell function has been described before, whereas the reduction in B cells has not been reported earlier. The results may suggest an impaired humoral immunity and a minor reduction in cellular immunity in patients following blood transfusion.
A New Low‐Frequency Red Cell Antigen, ‘SARAH’
Authors: Stern D.A. , Hawksworth D.N. , Watt J.M. , Ford D.S.
Published: March 2009

There are more than 100 low‐frequency antigens (LFAs) which have been given International Society of Blood Transfusion (ISBT) numbers as members of systems, collections or the 700 series. In addition, there are a number of well‐known (to reference laboratories) unpublished LFAs. The presence of an LFA was suspected when 2 sera were found to react with a single example of K homozygous cells. Anti‐K reacting with K homozygotes was eliminated on testing with other KK cells. Testing of the reactive cell with antibodies to known LFAs and the reactive sera with cells known to carry LFAs failed to identify the specificity. A study on the family of the cell donor showed inheritance of the antigen in two generations. Further testing, which included immunoblotting and RFLPs, was carried out in Australia, the UK, Canada and the USA. By March 1993 all published LFAs had been excluded, and an application was made to the ISBT to have the antigen, SARAH, assigned a 700 series number. In April the number 700.052 was provisionally designated for this new antigen.
Type 2 T‐Cell‐Independent Murine Immune Response to the Human AB0 Blood Group Antigens
Authors: Néron Sonia , Lemieu Réal
Published: March 2009

The human AB0 blood group determinants are simple carbohydrate structures which are widely distributed in nature. Much work has been done on the structure of the A and B antigens but little is known on the regulation of anti‐A and anti‐B immune responses. To develop a model system, we have characterized the AB0 immunity of normal Balb/c mice and found a significant level of serum natural anti‐A but almost no anti‐B. This finding and the known IgM predominance among immune anti‐A produced in the Balb/c mouse indicate that the AB0 immunity of this mouse strain is comparable to the one of human blood group B individuals. Following immunization with human red blood cells, similar levels of anti‐A were produced in normal and athymic Balb/c mice showing that the anti‐A response is T cell independent. Furthermore, no anti‐A or anti‐B antibodies were produced in CBA/xid mice indicating a type 2 T‐cell‐independent immune response. These results may contribute to a better understanding of human AB0 responses and establish the mouse as a suitable model to study the immunobiology of AB0 antigens.
Tolerability and Kinetics of a Solvent‐Detergent‐Treated Intravenous Immunoglobulin Preparation in Hypogammaglobulinaemia Patients
Authors: Ebeling Freja , Baer Maija , Hormila Pirkko , Järventie Greta , Koistinen Pirjo , Kätkä Kalevi , Oksanen Kalevi , Perkkiö Mikko , Ruutu Tapani , Soppi Esa , Veijola Lea , Myllylä Gunnar
Published: March 2009

The tolerability and kinetics of a solvent‐detergent‐treated 6% intravenous immunoglobulin (IVIG) preparation were studied in 15 hypogammaglobulinaemia patients during 3–4 regular substitution infusions of 9–18 g, the mean dose being 359 mg/kg. The infusions were well tolerated, and the trough serum IgG levels achieved were comparable to two commercial IVIG preparations. The stepwise increase of the infusion rate up to 5 mg/kg/min and the use of this IVIG as a 12% solution were possible without serious adverse events in all the 6 studied hypogammaglobulinaemia patients. This greatly reduced the time needed for the infusions.
Carboxyhemoglobin Levels in Neonatal Immune Hemolytic Jaundice Treated with Intravenous Gammaglobulin
Authors: Ergaz Zivanit , Gross Ditsa , Bar‐Oz Benjamin , Peleg Ofra , Arad Ilan
Published: March 2009

In order to examine the effect of intravenous immunoglobulin (IVIG) on the rate of hemolysis in immune hemolytic hyperbilirubinemia, we measured the carboxyhemoglobin levels of 5 newborn infants who were subjected to IVIG treatment. The pretreatment rate of hemolysis, in the 5 patients with isoimmune hemolytic jaundice (3 patients with Rh hemolytic disease of the newborn and 2 patients with ABO hemolytic disease of the newborn), as reflected by caboxyhemoglobin levels was higher than the rate of hemolysis in normal newborn infants. In 4 out of the 5 patients treated with IVIG, there was a rapid decline (> 30%) of carboxyhemoglobin levels, a pattern which was different from that observed in normal newborn infants with no hemolytic jaundice and in 3 untreated patients with ABO hemolytic disease of the newborn. None of the treated patients required an exchange transfusion. Our preliminary results support the theory that the attenuation of jaundice observed following IVIG treatment in patients with immune hemolytic hyperbilirubinemia is caused, at least in part, by the reduction in hemolysis.