bluuebunny/crossref_metadata_2025 huggingface dataset:
Stabilization of Human Albumin by Caprylate and Acetyltryptophanate
Authors: Yu M.W. , Finlayson J.S.
Published: March 2009

Abstract. The thermal stabilization of human albumin by caprylate (CA) and acetyltryptophanate (AT) was studied by monitoring the formation of albumin polymer (defined as species larger than dimer) on the basis of its molecular size as well as its characteristic migration as α‐globulin. Heating 5% protein solutions of purified albumin monomer, Cohn fraction V, and fraction IV‐4 + V at 60 °C in 0.1 M sodium phosphate or 145 mM sodium chloride, pH 7.0, established the following order of stabilizer effectiveness: 4 mM CA + 4 mM AT ≅ 4 mM CA > 8 mM AT ≥ 2 mM CA > 4 mM AT. However, albumin was more thermally stable in the chloride medium. Raising the CA concentration above 4 mM provided little additional stabilization. The D‐ and L‐enantiomers of AT were equally effective, but 16 mM AT was needed to equal the effect of 4 mM CA. L‐Tryptophanate exerted only slight stabilization, even at 32 mM; D‐tryptophanate was even less effective. The albumin polymer level increased progressively with time at 60 °C in 2 mM CA or 4 mM AT, whereas in 4 mM CA it reached a plateau in 4–6 h. Acetone drying of albumin‐rich fractions was shown to remove nearly all endogenous fatty acid, rendering the protein thermally labile unless sufficient exogenous stabilizer(s) was added. Even in the presence of 145 mM sodium chloride and 4 mM CA + 4 mM AT or 4 mM CA, the stabilizing effect of endogenous fatty acid was still detectable.
Viral Hepatitis Markers in Thai Blood Donors
Authors: Seidl S. , Chandanayingyong D.
Published: March 2009

Abstract. 1,410 voluntary blood donors from Bangkok were investigated by radioimmunoassay for HBsAg, anti‐HBs, anti‐HBc, HBeAg, anti‐HBe and anti‐HAV. An HBsAg carrier state was observed in 10.7% of whom 42.1% were HBeAg positive and 40.2% had anti‐HBe. Antibody prevalences (except anti‐HBe) rose with increasing age. The occurrence of hepatitis markers did not vary significantly between donors living along the canals and those living in the Greater Bangkok area.
Malaria‐Induced Immune Thrombocytopenia
Authors: Sørensen P.G. , Mickley H. , Schmidt K.G.
Published: March 2009

Abstract. On return from Liberia, a previously healthy 36‐year‐old man showed signs of malaria accompanied by severe haemolysis and slight thrombocytopenia. We found evidence of a platelet‐associated IgG being responsible for the thrombocytopenia, inasmuch as the direct platelet suspension immunofluorescence test was strongly positive, the indirect immunofluorescence test and tests for drug‐dependent antibodies at the same time being negative. We suggest that autoimmunity may be a contributing mechanism for platelet destruction in acute malaria.
Evaluation of an Ion‐Exchange Procedure for Removal of Hepatitis Type B Contamination from Human Gamma Globulin Products
Authors: Zolton Raymond P. , Padvelskis John V.
Published: March 2009

Abstract. Utilizing a unique human plasma sample that contained extremely high concentrations of hepatitis B surface antigen (HBsAg), we were able in the laboratory to rapidly screen the binding efficiency for HBsAg of various ion‐exchange resins that can be used to purify gamma globulin product from human plasma. ‘Nonrigid’ resins like QAE‐Sephadex and DEAE‐Sephadex showed a high affinity for HBsAg by removing greater than 104 of applied challenge. Much lower binding was achieved by ‘rigid’ resins like DEAE‐Sepharose CL‐6B and DEAE‐Spherodex. Also examined were the usefulness of certain auxiliary procedures, special filters and resins, that offer the potential to remove HBsAg. We failed to find any significant removal of the HBsAg, and in some cases large losses of the gamma globulin product occurred. While encouraged by our findings, certain precautions are discussed concerning the conversion from large‐scale production of gamma globulin by the conventional alcohol method to one utilizing chromatographic techniques.
Carbohydrate‐Specific Antibodies in Normal Human Sera
Authors: Lalezari Parviz , Jiang An Fu
Published: March 2009

Abstract. IgG and IgM antibodies against several sugars have been characterized in sera of normal donors by passive hemagglutination and a quantitative hemagglutination inhibition test. These antibodies distinguish between the equatorial and axial OH groups at C2, C3, or C4 positions of the glycopyranose configuration, differences between the anomers, linkage types, changes in the primary alcohol group at C6, and OH substitution. In the examples of antibodies to mannose, galactose, and glucose investigated, specificities were usually directed against the β‐anomers. In disaccharides, the antibodies appeared to react only with 1 of the 2 sugar subunits, but unlike monosaccharides, the glycosidic linkages also seemed to be a part of the reaction site. Thus, the reacting moiety in gentiobiose was β‐D‐glucopyra‐nosyl with 1→6 linkage, in cellobiose was β‐D‐glucopyranosyl with 1→4 linkage, in melibiose was α‐D‐galactopyranosyl with 1→6 linkage, and in lactose the reaction was directed against β‐D‐galactopyranosyl with 1→4 linkage. In the maltose‐dependent hemagglutination, α‐D‐glucose appeared to be the main reaction site. ManNAc exemplified the specificity determined by OH group substitution. Antibody to D‐Fucose represented example of specificity evolving from substitution of the primary alcohol.
A Rare Example of Weakened Expression of the Kell (K1) Antigen1
Authors: Kline William E. , Sullivan Carolyn M. , Bowman Robert J.
Published: March 2009

Abstract. Random typing of donor units revealed a donor whose red blood cells gave very weak reactions with all anti‐Kell (anti‐K1) antisera. All other Kell‐system antigens (including Kx) and antigens of other red‐cell systems tested were normal. This weak K1 antigen was present in four generations of the donor's family. The red‐cell morphology is normal in all family members with the weakened K1 antigen. A consistent hypothesis is that the weak antigen is inherited through a variant allele at the Kell locus and represents a quantitative, not a qualitative, change.
Effect of the Binding of Anti‐Zwa Antibodies on Platelet Function
Authors: van Leeuwen E.F. , Leeksma O.C. , van Mourik J.A. , Engelfriet C.P. , von dem Borne A.E.G. Kr.
Published: March 2009

Abstract. Anti‐Zwa antibodies can induce a Glanzmann‐like platelet dysfunction in normal Zwa‐positive platelets. Although a normal shape change and release reaction was always recorded in response to adenosine diphosphate and collagen, the aggregation on these inducers could be completely inhibited by anti‐Zwa antibodies. Analogous to the platelets from patients with Glanzmann's thrombasthenia, normal platelets sensitized with anti‐Zwa did not associate with fibrinogen upon exposure to adenosine diphosphate. Our data indicate that the binding site for fibrinogen is closely associated with the Zwa‐antigenic determinants on the platelet membrane glycoproteins and thus with glycoprotein IIIa which is known to carry Zwa.
A Mouse Monoclonal Antibody with Anti‐A,(B) Specificity Which Agglutinates Ax Cells
Authors: Moore S. , Chirnside A. , Micklem L.R. , McClelland D.B.L. , James K.
Published: March 2009

Abstract. A hybridoma (ES‐15) was obtained by fusing the NS‐1 cell line with spleen cells from a mouse immunised with soluble blood group A2 substance. The cloned hybridoma culture supernatant was shown to contain an IgM class antibody which strongly agglutinates group A cells and weakly agglutinates group B cells. The serological specificity of this antibody is described as anti‐A,(B) in this report. The abilities of unconcentrated monoclonal anti‐A,(B), a commercial human polyclonal anti‐A,B (group O serum) and a commercial monoclonal anti‐A reagent to detect 15 examples of Ax cells were compared by both slide and tube techniques. Using a slide technique monoclonal anti‐A,(B) agglutinated 14 examples of Ax cells, human anti‐A,B 2 examples, while monoclonal anti‐A failed to detect any of the Ax cells tested. Similar differences in the reactivity of the three antibodies were observed using a tube technique. Data are also presented which show that a 1:1 (v/v) mixture of monoclonal anti‐A,(B) with a monoclonal anti‐B reagent is an effective replacement for human anti‐A,B in ABO grouping procedures.
Monoclonal Antibodies to an Immunoglobulin Allotype Marker Glm(f)
Authors: Bird P. , Lowe J. , Jefferis R. , de Lange G. , van Loghem E. , Ling N.R.
Published: March 2009

Abstract. The properties of a group of mouse monoclonal antibodies (McAbs) specific for the human Glm(f) allotype marker on immunoglobulin G are described. The specificity of all 5 McAbs was anti‐Glm(f) in haemagglutination assays detecting the Glm(f) determinant on 6 ng of purified Glm(f) paraproteins. A high dilution (≥1/104) could be used for the majority of McAbs in this assay. In Elisa the Glm(f) marker could be detected in homozygote Glm(f+z‐) sera at a serum dilution of ≥ 1/104. In Elisa assays the Glm(f) specificity was lost when IgG was bound directly to polystyrene but was restored when IgG was bound via anti‐human IgG to the polystyrene plate. A possible conformational change in IgG to account for this loss of specificity is discussed. It is expected that these McAbs with their high titre and increased sensitivity over conventional Gm antisera will allow more detailed analysis of the Gm marker system.
Influence of IgG Subclasses on Automated Anti‐D (Rh0) Quantification
Authors: Boudart D. , Rivat‐Peran L. , Brossard Y. , Lopes P. , Yviquel M.
Published: March 2009

Abstract. Anti‐D quantification by both an automated Polybrene method and an automated trypsin‐albumin‐dextran (TAD) method gave discrepant results in certain cases. These discrepancies, expressed as the polybrene TAD ratio of reactivity (PTR), were related to the IgG subclass of anti‐D. Anti‐D of the IgG3 subclass showed a higher PTR than IgG1 (0.94 vs 1.65). No difference was shown between Glm(1) and Glm(3) (0.93 and 0.95, respectively) or between G3m(11) and G3m(21) (1.40 and 1.81, respectively) allotypes. The simultaneous use of our automated Polybrene and TAD methods provides information about the anti‐D subclass composition.
Isolation of Fab and Fc Fragments from a Plasmin‐Treated Human Immunoglobulin by High‐Speed Gel Filtration on TSK G3000SW and G3000SWG
Authors: Watabe S. , Takada K. , Shitori Y. , Hashimoto K.
Published: March 2009

A method is described for isolating intact 7S IgG, Fab and Fc fragments from a plasmin‐treated immunoglobulin by high‐speed gel filtration on a TSK G3000SW or G3000SWG column. The isolated 7S IgG, Fab and Fc fragments reacted with antihuman IgG, Fab and Fc antiserum, respectively, in both Ouchterlony's double immunodiffusion and immunoelectrophoresis. Each of the Fab and Fc fragments formed a single precipitation line, demonstrating their homogeneity. Anticomplementary activities of intact 7S IgG, Fab and Fc fragments were 2.5,≤10 and 2.5 mg protein/ml to inhibit 2 units of CH50, respectively, and the diphtheria antitoxin contents were 2.1, 6.0 and 1.5 IU/150 mg protein, respectively. The molecular composition of plasmin‐treated immunoglobulin as determined by gel filtration on a TSK G3000SW column was as follows: 7S IgG 38.0±0.2%; Fab 42.6±0.1% and Fc fragment 19.3±0.1%.
Protective Effects of Human Gamma‐Globulin Preparations against Experimental Aerosol Infections of Mice with Bordetella pertussis
Authors: Imaizumi Atsushi , Suzuki Yoji , Sato Hiroko , Sato Yuji
Published: March 2009

The protective effects of three types of pooled human gamma‐globulin preparations (intact: GG, S‐sulfonated: GGS, and pepsin‐treated: GGP) for intravenous use against experimental aerosol infection of mice with Bordetella pertussis have been evaluated. The gamma‐globulin preparations GG and GGS showed significant protective activity whereas GGP did not, as evaluated by survival numbers, body weight gains and suppression of leucocytosis. GG and GGS but not GGP also possessed neutralizing activity against leucocytosis‐promoting and histamine‐sensitizing activities of pertussis toxin (PT). Evaluation of protective activities of GG, GGS and GGP prepared from rabbit gamma‐globulin, highly immunized with PT and not containing any anti‐F‐HA (filamentous haemagglutinin) or agglutinin antibodies, demonstrated that anti‐PT‐GG and anti‐PT‐GGS but not anti‐PT‐GGP protected against experimental infection by B. pertussis. These studies showed that the protective activity was correlated with anti‐PT titre but that the Fc portion of the gamma‐globulin molecule is necessary for actual protection against whooping cough by B. pertussis.
Evidence for Sialylated Type 1 Blood Group Chains on Human Erythrocyte Membranes Revealed by Agglutination of Neuraminidase‐Treated Erythrocytes with Waldenström's Macroglobulin IgMwoo and Hybridoma Antibody FC 10.2
Authors: Picard Jean K. , Loveday Danuta , Feizi Ten
Published: March 2009

Haemagglutination studies have been performed with untreated and neuraminidase‐treated human erythrocytes of the three Lewis antigen types Le(a‐b‐), Le (a+b‐) and Le(a‐b+) using two monoclonal antibodies, IgMwoo and FC 10.2, which were previously shown to recognize the type 1 based blood group chains: Galβl→3GlcNAcβl→3Galβl→4Glc/GlcNAc (for explanation of abbreviations see table IV legend). Both antibodies behaved as cold agglutinins with neuraminidase‐treated but not with untreated erythrocytes of the three Lewis antigen types. Neuraminidase‐treated erythrocytes of i antigen type were similarly agglutinated. This haemagglutination was specifically inhibited by the type 1 based milk oligosaccharide lacto‐N‐tetraose. Thus, there is strong evidence for the occurrence of sialylated type 1 chains on human erythrocyte membranes of I and i antigen types. In addition, evidence for the presence of type 1 chains which are both sialylated and fucosylated was obtained by (1) haemagglutination of Le(a+b‐) erythrocytes with the monoclonal antibody 19.9; (2) increased haemagglutination of neuraminidase‐treated erythrocytes with anti‐H antibodies of Bombay serum; (3) increased haemagglutination of neuraminidase‐treated Le(a+b‐) cells with anti‐Lea antibodies, and (4) the appearance of Lea antigen activity on neuraminidase‐treated erythrocytes of Le(a‐b+) type.
HBeAg/Anti‐HBe Determination with a New Monoclonal Immunoradiometric Assay
Authors: Pizzocolo G. , Giammatteo D. , Barbieri U. , Boniolo A. , Callegaro L. , Palla M. , Albertini A.
Published: March 2009

Abstract. The performance of two assays for the HBeAg/anti‐HBe system associated with hepatitis B virus infection, using monoclonal (EBK‐Sorin) and polyclonal (HBe‐Abbott) antibodies, has been compared. The results on a random population (1,000 samples) demonstrated for the monoclonal reagent a higher sensitivity, without any loss in specificity, and with the further advantage of the use of lower radioactivity levels. The proportion of positives and negatives obtained with the two kits was found to remain unchanged in the case of HBeAg, while a markedly larger percentage of positives (7% higher) was detected for anti‐HBe using the monoclonal antibodies.
Biochemistry of Whole Blood in Poly(Ethylene‐Co‐Ethylacrylate) Experimental Blood Containers1
Authors: Horowitz Bernard , Rowe Arthur W. , Stryker Martin H. , van der Sande Johan , Waldman Alan A. , Wong Show‐Chu , Gass Jerald D. , Woods Kenneth R.
Published: March 2009

Abstract. The biochemical status of whole blood stored in containers fabricated of ethylene ethylacrylate (EEA) film was monitored at several times during 4 weeks of storage at 4°C. Fifteen biochemical indicators were studied to reflect on erythrocyte integrity, cellular metabolism, plasma protein stability, and microaggregate formation. Comparison to storage in polyvinyl chloride (PVC) containers was made by distributing aliquots from each unit of blood among the containers being compared. Whole blood in EEA developed significantly higher levels of plasma hemoglobin, erythrocyte osmotic fragility, and D‐glycerate‐2,3‐diphosphate (2,3‐DPG), and somewhat greater glucose utilization, lactate production, and pH. These biochemical differences were not of great magnitude and the data suggest that EEA containers are compatible with the storage of whole blood.
Stabilization of Red Blood Cells by the Plasticizer, Diethylhexylphthalate1
Authors: Horowitz Bernard , Stryker Martin H. , Waldman Alan A. , Woods Kenneth R. , Gass Jerald D. , Drago James
Published: March 2009

Abstract. The red blood cells of blood stored in containers made of polyvinylchloride (PVC) film are osmotically more stable and lose on average about 1/3 less hemoglobin than when blood is stored in another plastic [poly‐(ethylene‐co‐ethyl acrylate); EEA]. The stability of uniform volumes of stored red blood cells varies directly with PVC surface area, whereas changes in EEA surface area have comparatively little or no effect. PVC contains high concentrations of the plasticizer, diethylhexylphthalate (DEHP), known to migrate into blood and to have a high potential for toxicity. To determine if DEHP could be the red cell stabilizing agent in PVC, whole blood was stored in containers made from EEA into which was incorporated varying amounts of DEHP. Incorporation of DEHP into EEA significantly reduced erythrocyte osmotic fragility (p = 0.01). The degree of reduced fragility correlated with the level of DEHP in the cell phase implicating DEHP in PVC containers as the stabilizing agent for red cells.
Li Cold Agglutinin: A Further Antibody Recognizing Sialic Acid‐Dependent Antigens Fully Expressed on Newborn Erythrocytes
Authors: Roelcke D.
Published: March 2009

Abstract. The cold agglutinin anti‐Li recognizes a sialic acid‐dependent antigen fully expressed only on newborn red cells. Although it resembles anti‐i in all aspects except for its nonreactivity with neuraminidase‐treated red cells, it is entirely different from anti‐i, since sialic acid is not involved in i antigenic determinants.
A Two‐Stage Immunoradiometric Assay with 125I‐Staphylococcal Protein A for the Detection of Antibodies and Complement on Human Blood Cells1
Authors: Salama A. , Mueller‐Eckhardt C. , Bhakdi S.
Published: March 2009

Abstract. A sensitive two‐stage immunoradiometric assay (IRMA) using unlabeled specific antiglobulin sera followed by binding of radiolabeled staphylococcal protein A has been developed for detection of human IgG, IgM, IgA and complement C3 on human blood cells. In an experimental model system, purified human immunoglobulins (Ig) were coupled onto red blood cells, platelets, and leukocytes. The Ig‐coated red blood cells were analyzed in parallel by three methods: the radioimmune antiglobulin test with radiolabeled anti‐Ig; the radiolabeled staphylococcal protein A test, and the IRMA. Of the three methods evaluated, the latter was found to be sensitive and the easiest to perform. The applicability of the IRMA was established by investigating a group of 14 selected patients with autoimmune warm type hemolytic anemia. We observed that the IRMA detected cell‐bound antibodies (IgG and/or IgM) in several cases where conventional assays yielded negative results.
Use of Synthetic Antigens with the Carbohydrate Structure of Asialoglycophorin A for the Specification of Thomsen‐Friedenreich Antibodies1
Authors: Höppner W. , Fischer K. , Poschmann A. , Paulsen H.
Published: March 2009

Abstract. The panagglutination phenomenon described by Thomsen and Friedenreich (TF) is due to the reaction of naturally occurring TF‐antibodies with the carbohydrate group β‐D‐Gal‐(1–3)‐D‐GalNAc of desialylated glycophorin A, the major glycoprotein component of the erythrocyte membrane. The specificity of human TF‐antibodies reacting with this disaccharide was investigated by hemagglutination inhibition assay and radioimmunoassay using various synthetic oligosaccharides and neoglycoproteins as well as asialoglycophorin A. The results indicate that TF‐antibodies represent a heterogeneous mixture of carbohydrate‐specific antibodies. The disaccharide β‐D‐Gal‐(1–3)‐D‐GalNAc is the common structure recognized by all TF‐antibodies. However, the conjugation mode of the carbohydrate to the carrier protein is important for defining the specificity of different subpopulations of TF‐antibodies. The immunological reaction depends on the configuration of the glycosidical linkage as well as on the chemical nature of the aglycon, which is coupled to the disaccharide. These findings suggest that the heterogeneity of natural TF‐antigens is due to the wide distribution of the carbohydrate structure β‐D‐Gal‐(1–3)‐D‐GalNAc. The characterization of TF‐ or TF‐like antibodies directed to particular natural TF‐antigens (e.g. asialoglycophorin A, tumor TF‐antigens, glycolipids, bacterial antigens) requires TF‐analogues, which contain the additional molecular regions together with the TF‐disaccharide. These structures, apart from the TF‐hapten, are obviously important for defining the immunodeterminant group of TF‐antigens of different origin.
Quantitation of Immunoglobulin Classes and Subclasses in Anti‐Rh (D) Antibodies1
Authors: Mattila Petri S. , Seppälä Ilkka J. T. , Eklund Jarl , Mäkelä Olli
Published: March 2009

Abstract. The isotype profiles of anti‐Rh (D) antibodies were measured from 16 serum samples with a solid‐phase radioimmunoassay that employed monoclonal anti‐isotype antibodies. These antibodies had been standardized in another solid‐phase system with the aid of monoisotypic antibodies (e.g., anti‐tetanus toxoid of isotype IgG4), and this permitted calculations of the relative concentrations of different isotypes in each anti‐D population. Of the heavy‐chain isotypes, only IgG1 and IgG3 were found in anti‐Rh (D) antibodies. The share of IgG1 varied from 100% (2 sera) to 0% (1 serum) of detected units, and the mean proportion was 73.8%. The remainder was IgG3 (mean share 26.2%). The proportions of kappa and lambda immunoglobulins varied greatly in individual sera, in three sera only kappa Ig and in one serum only lambda Ig could be detected.
Whole Blood Immunoadsorption of Anti‐A or Anti‐B Antibodies1
Authors: Bensinger William I. , Buckner C. Dean , Clift Reginald A.
Published: March 2009

Abstract. Immunoadsorption columns capable of specifically removing antibodies show promise as an alternative treatment to plasma exchange. Most columns are designed to be perfused with plasma and it is necessary to use a plasma separation device (filtration or centrifugation) for the procedure. This increases the cost and complexity of the immunoadsorption procedure. We have adapted immunoadsorption columns capable of removing anti‐A or anti‐B red cell antibody from plasma to a whole blood perfusion system by coating the silica with a thin layer of collodion and incubating with albumin (1%). Patients were given heparin, 25 units/kg intravenously, followed by a continuous infusion of 3% citrate and heparin, 10 units/ml, to achieve a ratio of 1:16–1:20 (anticoagulant to whole blood) before delivery through the columns. Effective and specific removal of anti‐A or anti‐B antibody was achieved with this technique. Effects on coagulation parameters and the complement system were acceptably small.
Blood Donors Positive for HBsAg and Negative for Anti‐HBc Antibody
Authors: Couroucée Anne‐Marie , Drouet J. , LeMarrec N. , Drouet A. , Soulier J.P.
Published: March 2009

Abstract. From January 1980 to December 1983,989,907 blood donations were screened for HBsAg. A total of 1,345 was found HBsAg positive; 33 of whom were negative for anti‐HBc (2.45%); in 10 cases HBeAg was present at a low level along with HBsAg. 2 of the 33 subjects were lost to follow‐up. Late serum samples were available for 28 blood donors, and clinical and biological data only were known for the 3 others. An evolution of HBV markers was observed in each case; anti‐HBc antibody became positive in 24 subjects (the 4 remaining subjects had a very short follow‐up); HBeAg was positive at a high titer in 18 subjects and seroconversion to anti‐HBe was observed in 7 individuals; HBsAg concentration increased in 21 subjects from 2 to 4,000 times and decreased in the 7 others. Seroconversion to anti‐HBs was observed in 8 individuals. According to these serological, biological and clinical findings, different outcomes were observed: in 6 subjects too short a follow‐up was available to allow appropriate classification, but there was evidence to suggest all probably developed clinical hepatitis; in 6 subjects asymptomatic hepatitis with rapid loss of HBsAg occurred; in 2 subjects asymptomatic hepatitis with the persistence of HBsAg at a low level and seroconversion to anti‐HBe occurred; and in 17 subjects clinical hepatitis developed. These observations establish that most of the 33 blood donors were infectious at the time of their blood donation, and that anti‐HBc antibody screening must not replace HBsAg screening for blood donations.
Rapid Purification of Anti‐I and Anti‐i Cold Antibodies by Affinity Chromatography
Authors: Glick Joseph , Oppenheim Joel D.
Published: March 2009

Abstract. A method is described for the rapid purification of serologically active high titer anti‐I and anti‐i cold antibodies from the sera of patients with chronic cold agglutinin disease (CCAD). The purification procedure is based on thermal affinity chromatography, using desialated orosomucoid (α1‐acid glycoprotein)‐Sepharose 4B conjugated beads. The nature of the interaction between the cold agglutinins (CA) and the desialated orosomucoid is unknown. Inhibition studies, however, revealed that the cold hemagglutinating activities of all the anti‐i sera were inhibited by desialated orosomucoid while only 1 out of 4 of the anti‐I sera was similarly affected. Anti‐I or anti‐i antibodies were separated from whole sera in 7 out of 7 samples with a recovery in most cases of 100% of the cold hemagglutinating activity. The resultant products were purified monoclonal IgM fractions which could react with anti‐χ and anti‐μ. but not with anti‐λ, sera. The homogeneity, purity and specificity of all preparations were confirmed by immunodiffusion analysis against purified I and i blood group antigens isolated from human erythrocyte membranes, zonal and right‐angle electrophoresis and hemagglutination or hemagglutination inhibition studies.
A Diffusable Factor in Scottish Coagulation Factor Concentrates Can Inhibit Lymphocyte Transformation in vitro
Authors: McDonald Carolyn , Jackson Vivienne , Kilpatric David C. , Yap Peng Lee , Prowse Christopher
Published: March 2009

Abstract. Coagulation factor VIII and IX concentrates produced in Scotland inhibited human lymphocyte transformation induced by lectins or the recall antigen, purified protein derivative of tuberculin (PPD). Although concanavalin A could bind factor VIII directly and PHA could bind factor IX directly, most of the inhibition was not due to direct clotting factor – lectin interaction nor to simple toxicity. Most of the inhibitory activity from both clotting factors could be removed by dialysis. A similar degree of inhibition was observed when the buffers used to prepare the concentrates were substituted for the concentrates themselves, and a comparable concentration of sodium citrate was also found to be inhibitory. Coagulation factor VIII partially purified by gel filtration was not found to have an appreciable effect on lymphocyte transformation in vitro.
What is the ‘Optimum’ Optimal Additive Solution?
Authors: Napier J.A.F. , Ashford P.R. , Hayward M.M.
Published: March 2009

Spontaneous formation of clotted material occurs occasionally in all stored red cell units, but larger amounts have been noted in SAG‐M optimal additive suspensions. These can be substantially reduced by substitution of the SAG‐M solution by a SAG‐M/CPDA‐1 mixture suggesting that there is insufficient citrate in SAG‐M red cell suspensions to completely inhibit coagulation of the residual plasma.
Two Further Examples of the Low‐Frequency Antigen Rea (Reid)
Authors: Rowe G.P. , Bowell P.
Published: March 2009

A positive direct antiglobulin reaction of a baby's cells, found during antenatal testing at Cardiff, was caused by a low‐frequency antigen which has been identified as Rea (ISBT No. 700019). Investigations with this, only the second example, anti‐Rea revealed another Re(a+) propositus in testing 6,635 random donors at Oxford; no further positive was found in 4,770 random donors at Cardiff. Study of the families of these two propositi, the first to be identified since the original Canadian family, confirmed that Rea was inherited as an autosomal Mendelian dominant. The families showed that Rea is not controlled by the ABO, Kell or Au loci and confirmed that it was not controlled by MNSs and was not X‐ or Y‐borne.
Effects of Oxygen on Red Cells during Liquid Storage at +4°C
Authors: Högman Claes F. , De Verdier Carl‐Henric , Ericson Åke , Hedlund Kenneth , Sandhagen Bo
Published: March 2009

Red cells collected in CPD and suspended in SAGM medium were stored in plastic (PVC) containers for 42 days at +4°C. Comparison was made between aerobic storage (normal air exposure) and anaerobic storage (exposure to nitrogen gas). The air‐exposed units showed a strong increase in pO2 and oxygen saturation as a result of oxygen penetration into the bags from outside. This resulted in a decrease in ATP and adenylate energy charge, a slower metabolization of adenine and hypoxanthine to AMP and IMP, respectively, and a faster decrease in red cell fluidity. To explain the findings it is concluded that aerobic storage causes an increased need of high‐energy phosphate groups, possibly used for replacement of the phospholipid membrane bilayer or in repair of phosphate bonds in the cytoskeleton. It is further proposed that a slight formation of hydrogen peroxide from free oxygen radicals moderately increases the oxidation of reduced (GSH) to oxidized (GSSG) glutathione and slightly enhances the need for reduced nicotinamide‐adenine dinucleotides mainly provided by increased flux through the pentose phosphate shunt.
Correlation of Immune Defects in Hemophilia with HTLV‐III Antibody Titers
Authors: Daniel V. , Opelz G. , Schäfer A. , Schimpf Kl. , Wendler I. , Hunsmann G.
Published: March 2009

Abstract. Of 170 hemophilia patients, 22% had high‐titer, 29% had low‐titer, and 49% had no antibodies against HTLV‐III. The strength of HTLV‐III antibodies was correlated significantly with a decreased OKT4/T8 ratio (p < 0.0005), decreased in vitro response to pokeweed mitogen (p < 0.025), and elevated serum neopterin (p < 0.05) and serum IgG (p < 0.0005). The fraction of patients with abnormal immunological findings was consistently greater among patients with high‐titer than among patients with low‐titer HTLV‐III antibodies. Testing these immunological parameters may be useful for monitoring the breakdown of immune functions leading to AIDS.
A Study of IgE in Immunoglobulin Preparations for Intravenous Administration
Authors: Paganelli R. , Quinti I. , D'Offizi G. P. , Papetti C. , Cabello A. , Aiuti F.
Published: March 2009

Abstract. We have studied the IgE content of different lots of commercial intravenous IgG preparations used in our day hospital as replacement therapy for patients with agammaglobulinemia. Two enzymatic methods were used: Phadezym and FAST. The average amounts of IgE detected in Endobulin and Sandoglobulin lots were very high with respect to serum values in the general population. In some lots we found low titers of specific IgE (RAST class 1) to house dust mite, rye grass and cow's milk. We also found in most preparations the presence of IgG anti‐IgE which are usually present in sera from atopic patients. Despite a low post‐infusion increase of serum IgE and the absence of sensitization or adverse reactions observed, a careful selection of donors with normal IgE levels may need to be recommended to manufacturers in the future.
Inhibition of Immune Phagocytosis by Human Sera with HLA A, B, C and DR but Not with DQ or EM Type Reactivity
Authors: Neppert Jürgen , Pohl Elisabeth , Mueller‐Eckhardt Christian
Published: March 2009

Abstract. 613 sera from pregnant women were investigated for inhibition of immune phagocytosis (IPI) by monocytes exposed to anti‐D (Rh)‐sensitized human red blood cells. IPI was detected in 42 (26%) of 165 and in 78 (17%) of 448 sera assessed against monocytes from 15 or 5 panel donors, respectively. All IPI‐positive sera reacted in an allotypic pattern. Eight IPI‐positive sera tested with autologous monocytes were found to be nonreactive. Upon comparative analysis of 448 sera, a significant correlation was found between the specific patterns of IPI and lymphocytotoxicity by HLA antibodies. In addition, 13 sera (4%) were positive for IPI, but not for lymphocytotoxicity. Twenty IPI‐positive sera were tested by indirect immunofluorescence against platelets and T lymphocytes and revealed IgG antibodies in 16 and 11 sera, respectively. IPI activity could be removed from 8 out of 10 positive sera by platelet pool absorption. While virtually all sera exhibiting HLA, A, B or C‐like activity (cytotoxicity with all cells) or HLA DR‐like activity (exclusive cytotoxicity with B lymphocytes and monocytes) were IPI‐positive, IPI was infrequently observed with sera containing HLA DQ‐like activity (cytotoxicity with B lymphocytes only). IPI was also rarely seen with sera cytotoxic only to monocytes and not at all with sera containing antibodies against endothelial/monocyte antigens. We conclude that IPI is caused by cytotoxic as well as noncytotoxic HLA A, B, C, DR‐specific antibodies. This effect may bear significance for the maternal immune response against fetal antigens and may be useful for pretransplant histocompatibility testing.
An Autoanalyzer Test for the Quantitation of Platelet‐Associated IgG1
Authors: Levitan Nathan , Teno Richard A. , Szymanski Irma O.
Published: March 2009

Abstract. A new quantitative antiglobulin consumption (QAC) test for the measurement of platelet‐associated IgG is described. In this test washed platelets are incubated with anti‐IgG at a final dilution of 1:2 million. The unneutralized fraction of anti‐IgG remaining in solution is then measured with an Autoanalyzer and soluble IgG is used for calibration. The dose‐response curves depicting the percent neutralization of anti‐IgG by platelets and by soluble IgG were compared in detail and found to be nearly identical, indicating that platelet‐associated IgG can be accurately quantitated by this method. The mean IgG values were 2,287 molecules/platelet for normal adults and 38,112 molecules/platelet for ITP patients. The Autoanalyzer QAC test is a sensitive and reproducible assay for the quantitation of platelet‐associated IgG.
Generation and Degradation of Fibrinopeptide A in Stored Platelet Concentrate
Authors: Bode Arthur P. , Miller David T.
Published: March 2009

Abstract. The levels of fibrinopeptide A (FPA) were measured in samples from stored platelet concentrates (PC) by radioimmunoassay. In 27 standard, citrated PC, the mean FPA was 13.6 ng/ml, which is elevated 5–6 ′ over background levels. This value did not change significantly over a 7‐day storage period. Addition of PGE‐1 and theophylline resulted in higher initial levels of FPA (18.0 ng/ml) and a pronounced rise during the storage period (to 43.4 ng/ml by day 10). In contrast, addition of a thrombin inhibitor, D‐phenylalanyl‐L‐prolyl‐L‐arginine chloromethyl ketone – 2 HCl (PPACK) or hirudin, resulted in lower initial levels of FPA relative to standard, citrated PC and a slow increase over time. Introduction of exogenous FPA into citrated PC resulted in a predicted elevation of FPA levels followed by a rapid loss of immunoreactivity (t1/2 = 18 h). Addition of PPACK did not affect this fall‐off. However, PC prepared and stored in the presence of PGE‐1 and theophylline showed a much slower fall‐off of exogenous FPA (t1/2 = 38 h). These data indicate that FPA levels in samples from citrated PC represent a dynamic balance between generation and degradation processes and, thus, the data above underestimate the amount of thrombin activity present in stored, citrated PC.
Standardization of the Processing of Human Bone Marrow for Allogeneic Transplantation
Authors: Gilmore M. J. M. L. , Prentice H. G.
Published: March 2009

Abstract. Allogeneic bone marrow transplantation has been undertaken within this centre on 62 patients with acute or chronic leukaemia. Employing the standard separation protocol described, all bone marrows were processed on the 2991 Blood Cell Processor to isolate the ‘buffy coat’ cells and subsequently the mononuclear cell component using a density separation medium of Ficoll‐metrizoate. Following the mononuclear cell separation, the cells were identified for their T lymphocyte component using a combination of two murine monoclonal antibodies, MBG6 reacting with a pan T cell antigen (CD6) and RFT8 detecting the ‘cytotoxic/supressor’ cell antigen (CD8). The numerical results for nucleated cells, red blood cells, T lymphocytes and colony forming units‐granulocyte macrophage are presented.
Reactions of Murine Monoclonal Antibodies to Blood Group MN Antigens1
Authors: Rubocki Ronald , Milgrom Felix
Published: March 2009

Abstract. BALB/c mice were immunized by intraperitoneal injections with human blood group substances; 4 mice with M and 4 with N. Immune spleen cells were fused with murine myeloma cells X63‐Ag‐8.653. Two clones secreting monoclonal anti‐M and seven secreting monoclonal anti‐N were identified. All antibodies were of IgG1 subclass and had k light chains. Three clones have been maintained that produced antibodies useful for typing purposes: anti‐M, A09 originating from a mouse injected with M, anti‐N, AH7 originating from a mouse injected with N, and anti‐N BO10 originated from a mouse injected with M substance. In typing 370 erythrocyte samples, monoclonal reagents gave identical results with commercial anti‐M or anti‐N typing sera of rabbit origin. Significantly, anti‐N reagent AH7 obtained by immunization with N substance showed serological differences from anti‐N reagent BO10 obtained by immunization with M substance in that AH7 had apparently higher avidity to N specificity on glycophorin A of N erythrocytes than BO10, whereas BO10 showed higher avidity for ‘N’ specificity on glycophorin B of M and N erythrocytes than AH7. These two reagents showed also somewhat different patterns of reaction with enzymatically digested erythrocytes. This apparent serologic difference between N and ‘N’ specificities is at variance with current immunochemical data.
Comparisons of Pooled Polyclonal Rabbit Anti‐Human C3d with Four Monoclonal Mouse Anti‐Human C3ds
Authors: Chaplin Hugh , Monroe Martha C.
Published: March 2009

Abstract. Performance characteristics of pooled rabbit IgG polyclonal anti‐C3d are compared with one mouse IgM and three mouse IgG monoclonal anti‐C3d antibodies (MAs). IgG MA,s employed singly or in combination, failed to precipitate C3d; by contrast, IgM MA and polyclonal anti‐C3d precipitated C3d. Measurements of polyclonal anti‐C3d concentration by chemical means and by 125I‐C3d radioimmunoassay (RIA) agreed closely. RIA values were 50% of chemical measurement values for three of the four MAs. Use of sucrose density gradient ultracentrifugation to assess MA C3d/anti‐C3d molar combining ratios for soluble anti‐C3d/C3d was not possible because fast‐sedimenting multimeric C3d/anti‐C3d complexes did not form. Dissociation and competitive binding studies indicate that (1) two MAs haad substantially lower affinities than the other anti‐C3d antibodies, and (2) polyclonal anti‐C3d recognizes more C3d epitopes than are recognized by individual MAs. The results demonstrate antigenic complexity of C3d fragment and illustrate the difficulties of predicting individual MA performance based on prior experience with polyclonal antibodies.
Comparisons of Pooled Polyclonal Rabbit Anti‐Human C3d with Four Monoclonal Mouse Anti‐Human C3ds
Authors: Chaplin Hugh , Monroe Martha C.
Published: March 2009

Abstract. Labelled polyclonal IgG anti‐C3d and monoclonal IgM and IgG anti‐C3d antibodies (MAs) were employed at increasing antibody excess to measure the number of C3d molecules on human red blood cells (RBC) coated by complement in vitro and in vivo. Values for the number of C3d sites per cell determined with polyclonal anti‐C3d were at least 4‐fold higher than when MAs were used. The results suggest that the molar combining ratio for polyclonal anti‐C3d with a single RBC‐bound C3d fragment is more likely >4.0 than 1.0 as previously assumed. Antiglobulin agglutination studies compared polyclonal and monoclonal anti‐C3d antibodies against C3d‐coated RBC from 27 patients with autoimmune hemolytic anemia. All four MAs showed striking prozones, requiring their use over a 25‐fold higher range of dilutions than polyclonal anti‐C3d. Polyclonal anti‐C3d produced stronger agglutination reactions than any of the IgG MAs. Only the IgM MA produced agglutination as strong as, or stronger than, polyclonal anti‐C3d. While IgM MA always gave the strongest MA agglutination reactions, no consistent ranking of the three IgG MAs was observed. Agglutination was not enhanced when all IgG MAs were combined; addition of IgG MAs to IGM MA reduced the strength of agglutination seen with IgM alone, suggesting blocking of IgM binding by competing IgG anti‐C3d.
Fc Receptors for IgG1 and IgG3 on Human Mononudear Cells ‐An Evaluation with Known Levels of Erythrocyte‐Bound IgG
Authors: Zupańska Barbara , Thomson Eileen E. , Merry Anthony H.
Published: March 2009

Abstract. The Fc receptors on mononuclear cells were investigated by a rosette technique in which human erythrocytes were sensitized with a known number of molecules of anti‐Rh antibodies (IgG1 or IgG3). The number of IgG molecules was quantitated by a radiometric antiglobulin test. The present quantitative evaluation reveals that (1) there is a logarithmic relationship between the proportion of rosettes and the amount of erythrocyte‐bound immu‐noglobulin for both types of mononuclear cells, and for both subclasses; (2) similar percentage of rosettes can be obtained with fewer IgG3 than IgG1 molecules (about 1:4); (3) for a given number of erythrocyte‐bound immuno‐globulins a higher percentage of rosettes is observed with monocytes than with lymphocytes (ratios of about 3:1 for IgG1 and 5:1 for IgG3); (4) the minimum number of IgG3 molecules for adherence is 180–460 for monocytes, 520‐1,300 for lymphocytes, whilst for IgG1 the numbers are 1,180‐4,300 for monocytes and 3,400‐14,200 for lymphocytes; (5) the minimum levels of sensitization by alloantibodies for adherence should be detectable by the antiglobulin test.
Enrichment of Human Bone Marrow Aspirates for Low‐Density Mononuclear Cells Using a Haemonetics Discontinuous Blood Cell Separator
Authors: Raijmakers R. , de Witte T. , Koekman E. , Wessels J. , Haanen C.
Published: March 2009

Abstract. Isopycnic density floatation centrifugation has been proven to be a suitable technique to enrich bone marrow aspirates for clonogenic cells on a small scale. We have tested a Haemonetics semicontinuous blood cell separator in order to process large volumes of bone marrow with minimal bone marrow manipulation. The efficacy of isopycnic density floatation was tested in a one and a two‐step procedure. Both procedures showed a recovery of about 20% of the nucleated cells and 1–2% of the erythrocytes. The enrichment of clonogenic cells in the one‐step procedure appeared superior to the two‐step enrichment, first separating buffy coat cells. The recovery of clonogenic cells was 70 and 50%, respectively. Repopulation capacity of the low‐density cell fraction containing the clonogenic cells was excellent after autologous reinfusion (6 cases) and allogeneic bone marrow transplantation (3 cases). Fast enrichment of large volumes of bone marrow aspirates with low‐density cells containing the clonogenic cells by isopycnic density floatation centrifugation can be done safely using a Haemonetics blood cell separator.
Prevalence of Hepatitis B Surface Antigen in Northern Nigerian Blood Donors
Authors: Kulkarni A. G. , Aloowooja F. O. , Wayo G. B.
Published: March 2009

Abstract. 1,860 serum samples of blood donors from northern Nigeria were tested for the presence of the hepatitis B surface antigen. 8.9% of them were found to be positive. Significant differences in frequency were observed amongst various ethnic groups as well as ABO blood groups.
Blood Units for Patients with Irregular Blood Group Antibodies: Frequency and Supply
Authors: Frick T. , Schenker T. , Frey‐Wettstein M.
Published: March 2009

Abstract. Blood banks face significant problems supplying blood units compatible with erythrocyte antigens other than ABO and rhesus D. In this study the number of such units was evaluated between 1973 and 1982, and it was found that 1.5% of the total number of blood units were compatible with other antigens than ABO and rhesus D. Based on data of the number of units requested for each such antigen and the frequency of the antigen in the European population, a logistical system providing sufficient numbers of ‘antibody‐compatible’ blood units is proposed.
On the Specificity of Human Anti‐H Antibodies
Authors: Pendu J. Le , Lambert F. , Gérard G. , Vitrac D. , Mollicone R. , Oriol R.
Published: March 2009

Abstract. Anti‐H antibodies from 2 homogeneous subgroups of H‐deficient individuals (Bombay and Réunion phenotypes) were studied for their specificity with regard to synthetic H oligosaccharides. It appears that Bombay individuals synthesize large amounts of both anti‐H type 2 and anti‐H type 1 antibodies, whereas Réunion individuals may possess strong anti‐H type 1 antibodies, but have only weak anti‐H type 2 antibodies in their serum. This can be explained by the presence of small amounts of H‐type 2 determinants on the erythrocytes of Réunion individuals contrasting with the complete lack of H determinants on the red cells of Bombay individuals.
Assessment of Multimeric Structure and Ristocetin‐Induced Binding to Platelets of Von Willebrand Factor Present in Cryoprecipitate and Different Factor VIII Concentrates
Authors: López‐Fernández M.F. , López‐Berges C. , Corral M. , García‐Talavera J.R. , Borrasca A. López , Batlle J.
Published: March 2009

Abstract. The multimeric structure of von Willebrand factor (vWF) and its ristocetin‐induced binding to platelets, using a simple and very sensitive radiomonoclonal antibody‐labeled vWF method, was compared in normal plasma, single‐donor cryoprecipitate (CP) and five different antihemophilic factor (AHF) concentrates. All the AHF showed a lack of larger vWF multimers, an abnormal ‘triplet’ pattern, and much lower vWF binding to platelets than that of plasma or CP, vWF being the lowest for those with a lesser proportion of larger vWF multimers. These results suggest that the combination of vWF multimeric analysis and the radiomonoclonal‐labeled vWF method may be very useful in the assessment of AHF preparations.