bluuebunny/crossref_metadata_2025 huggingface dataset:
Assessment of Red Cell Autoantibodies in Autoimmune Hemolytic Anemia of Warm Type by a Radioactive Anti‐IgG Test1
Authors: Schmitz N. , Djibey I. , Kretschmer V. , Mahn I. , Mueller‐Eckhardt C.
Published: March 2009

To evaluate the applicability of a radioactive 125I‐anti‐IgG test (RIAT) for the detection of small amounts of IgG antibodies on red blood cells (RBC) of patients with autoimmune hemolytic anemia of warm type (AIHA), RBC of 125 patients were studied (AIHA, n=53; Coombs'‐negative AIHA, n=6; chronic cold agglutinin disease, n=7; non‐immune anemias, n=59). It was found that the RBC of all cases (33/33) with a positive direct IgG antiglobulin test (DAT‐IgG), but also 13 out of 20 patients with a negative DAT‐IgG, but detectable complement (C3/C4), and 4 out of 6 cases with Coombs'‐negative AIHA gave positive results in the direct RIAT. RBC‐associated IgG was higher in the DAT‐IgG positive group (n=33; x̄=8.1%) than in the DAT‐IgG negative group (n=26; x̄3.4%). There was no correlation between hypergammaglobulinemia and RBC‐associated IgG. The sensitivity of the indirect RIAT was not remarkably better as compared to the indirect antiglobulin test. The RIAT is valuable in the serology of borderline cases of AIHA.
Auto‐anti‐B in a Patient with Acute Leukaemia
Authors: McClelland W.M. , Bradley A. , Morris T.C.M. , Burmide P. , Robertson J.H.
Published: March 2009

The presence of an auto‐antibody with group B specificity is described for the first time in acute leukaemia. The antibody also shows certain unusual serological characteristics.
Delayed Serum Sickness‐Like Transfusion Reactions in a Multiply Transfused Patient
Authors: Avoy Donald R.
Published: March 2009

The clinical features and progress of a patient with a fatal myeloproliferative disease are reported. Her care required frequent transfusions of red cell products and components. These transfusions were followed, after varying intervals, by fever, arthralgia, myalgia, headache and pericarditis. These reactions could be avoided by transfusing plasma‐free products, and were ameliorated by systemic steroids. This patient, then, is the first reported case of delayed, serum sickness‐like transfusion reactions. During the course of her illness she was demonstrated to have antibodies to immunoglobulins. The role of these antibodies, if any, is uncertain.
Differential Coating of Human Red Blood Cells with C4 or C3 in a Low Ionic Strength Medium
Authors: Heier H. E. , Kornstad L. , Nordhagen R.
Published: March 2009

An EDTA‐containing, low ionic strength medium, pH 5.1, can be used for incubation of blood to obtain coating of red blood cells with C4 or C3. The coating achieved depends on incubation temperature (4 or 37°C) and on whether or not CaCl2 or MgCl2 is added to the medium. The optimal concentration of EDTA appears to be 2 mM under the present conditions. This is equimolar to the final concentration of the added CaCl2/MgCl2. The choice between CaCl2 and MgCl2 depends on whether blood without anticoagulant or ACD blood is employed. Coated cells keep satisfactorily for 5 weeks at 4°C in Alsever's solution as well as after freezing and thawing. C3‐coated cells can be converted to C3dcoated ones by incubation in normal, compatible, EDTA‐containing serum at 37°C.
Prothrombin Complex Concentrates for Clinical Use1
Authors: Chandra S. , Brummelhuis H.G.J.
Published: March 2009

Prothrombin complex concentrates prepared for clinical use are reviewed with emphasis on frequently observed adverse reactions such as viral hepatitis and thromboembolic complications. Preparation procedures and quality control are described briefly, and the use of these concentrates in the treatment of hemophilia A patients who develop factor VIII inhibitors is also described. The causes of and methods for the detection of potential thrombogenicity are discussed and suggestions which may result in a safer product are made.
Red Cell Preservation in Protein‐Poor Media: III. Protection Against in vitro Hemolysis
Authors: Högman Claes F. , Hedlund Kenneth , Sahlestrom Ylva
Published: March 2009

Red cells stored under blood bank conditions normally show less than 1% spontaneous in vitro hemolysis even after 5 weeks; larger hemolysis may be found if the cells are suspended and stored in a saline‐adenine‐glucose (SAG) solution with very little trapped plasma. Delay of the addition of the suspension medium, return of 25 ml plasma after a maximal plasma harvest, addition of mannitol 10–30 mmol.1‐1 to the suspension medium were alternative and effective ways of keeping the spontaneous lysis within normal limits. Mechanical traumatization (centrifugation or shaking) caused considerably more damage to the red cells when these were highly concentrated than when they were diluted. A cell suspension in SAG is a more suitable product for hemotherapy than strongly packed red cell concentrates.
An Investigation into the Relationships between Serum Levels of C3, C4 and CH50
Authors: Salkie M.L. , Kelker D.H.
Published: March 2009

Aliquots of 213 specimens were assayed for C3, C4 and CH50. A graphical presentation of the data was obtained by a block diagram. This showed a generally flat plane with good agreement to quoted normal ranges.

Only 2 of 68 specimens with C3 and C4 normal have a low CH50. If either the C3 or C4 is low then the CH50 may be normal or even high, but if both fractions are low then so is the CH50.

A low CH50 can occur with both C3 and C4 in the normal range or even higher. This implicates other complement factors as being deficient with a sufficiently high occurrence rate to require that the CH50 be kept as a routine laboratory procedure.
A New Miltenberger Class Detected by a Second Example of Anek Type Serum
Authors: Dybkjaer E. , Poole J. , Giles C.M.
Published: March 2009

An incomplete IgG antibody, detected in compatibility testing, had the same specificity as Anek serum, a type serum of the Miltenberger complex. The cells of the donor represent a new Mi class, VIII, by sharing determinants found on cells of Mi IV, VI and VII.
Red Blood Cell Cold Agglutinins and B Lymphocyte Cytotoxins in Breast Cancer Sera1
Authors: Dube Volker , Iwaki Yuichi , Anderson Byron , Terasaki Paul
Published: March 2009

Cytotoxic activity towards B lymphocytes in sera from individuals with breast carcinoma was related to red blood cell cold agglutinin levels and IgM values. The control group consisted of 32 females with a similar age distribution. Significant increases in red blood cell cold agglutinin levels were found in the group of cancer patients and the group with the histological category of mucin‐producing carcinoma with lymphocyte cytotoxins (p<0.025, respectively), in comparison to the control group and cancer patients without lymphocyte cytotoxins. Serum IgM concentrations were significantly increased in the group of cancer patients (p<0.025) and the group with mucin‐producing carcinoma (p<0.01) with lymphocyte cytotoxins, in comparison to cancer patients and controls without lymphocyte cytotoxins. The lymphocyte cytotoxins were completely or partially adsorbed by red blood cells in 58% of the positive sera. These results suggest that red blood cell cold agglutinins constitute a portion of cytotoxic anti‐B lymphocyte antibodies. At present it is unknown whether the lymphocytotoxic antibodies in breast cancer patients are of importance in immunologic host defenses.
A New Approach to Washing Red Blood Cells Frozen with a High Concentration of Glycerol in a Special Freezing Container1
Authors: Kurtz S.R. , Valeri D.A. , Gray A. , Lindberg J.R. , McMican A. , Blumberg N. , Valeri C.R.
Published: March 2009

We report here on a new approach to washing red blood cells frozen with a high concentration of glycerol in a special freezing container. The wash solution consists of a 150‐ml volume of 12% sodium chloride and 2 liters of 0.9% sodium chloride‐0.2% glucose‐25 mEq/l disodium phosphate. Both the Haemonetics Blood Processor 115 and the IBM Blood Processor 2991 have been used with this protocol, with similar results. The in vitro recovery of red blood cells frozen with 8.6M glycerol was 89%, and that of red blood cells frozen with 6.2M glycerol was 93%. The 24‐hour posttransfusion survival values averaged 88% for eight units of outdated‐rejuvenated previously frozen red blood cells washed by this protocol and stored at 4°C for 3 days before autotransfusion.
Plasma Exchange Combined with Cytotoxic Drugs and Lymphocytapheresis for Myasthenia Gravis
Authors: Valbonesi M. , Garelli S. , Zerbi D. , Forlani G. , Cornelio F. , Pelucchetti D.
Published: March 2009

20 patients with myasthenia gravis (MG), refractory to anticholinesterase and steroid therapy, underwent plasma exchange therapy combined with immunosuppressive drugs and lymphocytapheresis. In all these patients an apparent clinical improvement was obtained since their first exchange session. During a follow‐up of 8–18.5 months, a long‐lasting benefit was achieved in 16 patients even though 6 of these had a single recurrence shortly after their first apheretic cycle. 3 patients who achieved substantial improvement of their symptoms during plasmapheresis showed recurrence of weakness after each apheretic course was stopped. However, their clinical response to therapy tended to improve with the following courses, which would be consistent with the effects of immunosuppression. This study strongly suggests that plasma exchange combined with immunosuppressive drugs and lymphocytapheresis can bring about dramatic and sustained improvement of MG and may alter its natural history.
Elution of Granulocyte and Platelet Antibodies
Authors: Helmerhorst F.M. , van Oss C.J. , Bruynes E.C.E. , Engelfriet C.P. , von dem Borne A.E.G.Kr.
Published: March 2009

By lowering the pH of the medium, allo‐ and autoantibodies bound to granulocytes and platelets could be eluted from these cells and their presence in the eluate could subsequently be detected with normal donor granulocytes and platelets, respectively, in the immunofluorescence test and/or agglutination test. IgG‐ as well as IgM‐ and IgA‐containing antibodies were detectable in the eluates. Depending on the attraction mechanism between antigen and antibody, some antibodies were also eluted by dimethylsulphoxide at pH 9.5, ether and/or by heating at 56°C. By differential elution of antibodies from donor platelets, we demonstrated the simultaneous presence of platelet‐specific antibodies and HLA antibodies in the serum of a mother whose newborn suffered from transient neonatal thrombocytopenia.
Observations on the Anton Antigen and Antibody
Authors: Poole Joyce , Giles Carolyn M.
Published: March 2009

Five antisera with Anton specificity were studied serologically. The determinant is not sensitive to treatment by proteolytic enzymes (notably trypsin) and is present on cells of the recessive type of Lu(a‐b‐) phenotype. The Anton ‘para‐Lutheran’ antigen/antibody therefore seems to be associated with the Lutheran blood group system only by means of cells of the dominant type of the Lu(a‐b‐) phenotype which is controlled by In(Lu), a gene independent of the Lutheran locus.
Elution of Group‐Specific Substance A from RBC of Various Subgroups of A and Its Effect on the Agglutination of Ax RBC
Authors: Dodd B.E. , Wood N.J.
Published: March 2009

Experiments are described which demonstrate that when the Landsteiner heat elution technique is used to prepare eluates from group A cells, some A substance as well as anti‐A is released into the eluates. The concentration found is greatest from A1 RBC and least from Ax, suggesting a relationship with the number of antigen sites on RBC of the various subgroups. Further experiments have shown that the presence of this A‐specific substance in eluates accounts for the well‐know phenomenon that anti‐A from selected group O sera, when eluted from A1 RBC, will not agglutinate Ax even though the original serum showed an appreciable titre against Ax. The presence of the eluted antigen also suggests an explanation for the behaviour of antibody recovered from the red cells of various subgroups of A towards Ax.
Immunochemical Studies of Infectious Mononucleosis
Authors: Fletcher Mary Ann , Caldwell Karen E. , Latif Zuhair
Published: March 2009

Abstract. A glycoprotein was isolated from the membrane of the bovine erythrocyte by refluxing the acetone‐ and ethanol‐extracted stroma residue with 75% ethanol. The glycoprotein was purified by phosphocellulose chromatography, ethanol precipitation, lipid‐solvent extraction and DEAE chromatography. The glycoprotein appeared to have two serological determinants, both reactive with antibodies present in the sera of patients with infectious mononucleosis. One of the determinants is similar to the Paul‐Bunnell heterophile antigen found on sheep erythrocytes. It is dependent on carbohydrate, including sialic acid residues. Another specificity, seemingly not shared by sheep erythrocytes to any great extent, is resistance to neuraminidase and to alkaline borohydride treatment and thus it may be located either on the polypeptide portion of the molecule or on an alkali‐stable oligosaccharide. The purified glycoprotein comprises 73% amino acids. Carbohydrate components and their molar ratios were sialic acid (1.0): galactose (1.5): N‐acetylglucosamine (1.1): N‐acetylgalactosamine (0.5): mannose (0.1).
Improved Assay for Detection of Platelet‐Specific PlA1 Antibodies in Neonatal Alloimmune Thrombocytopenia
Authors: Mueller‐Eckhardt C. , Kayser W. , Förster Christa , Mueller‐Eckhardt Gertrud , Ringenberg C.
Published: March 2009

Abstract. A technique is described which permits the detection of platelet‐specific, noncomplement‐fixing (‘blocking’) P1A1 antibodies in serologically negative sera of mothers from children suffering from neonatal alloimmune thrombocytopenia. The technique consists of (1) antibody enrichment in eluates prepared from maternal serum and P1A1‐positive platelets, and (2) quantitation of antibodies in eluates by the platelet radioactive anti‐IgG test. With this assay, P1A1 antibodies were demonstrated in 6 out of 7 maternal sera, and anti‐A antibodies in one. It proved valuable also for longitudinal studies for periods up to 30 months after delivery.
Alloantibody‐Induced Impaired Neonatal Expression of a Red Blood Cell Antigen Associated with Maternal Alloimmunization
Authors: Ferguson S.J. , Blajchman M.A. , Guzewski H. , Taylor C.R. , Moulds J.
Published: March 2009

Abstract. Animals genetically capable of making certain gene products have been shown to have the production of such products completely or partially suppressed by exposure in utero or neonatally to antibodies specific for that gene product. This alloantibody‐induced depression of expression of a specific antigenic determinant has yet to be shown to occur in man. The McCoya red blood cell antigen is reported to be well developed at birth. However, 2 children born to mothers with high‐titer McCoya antibodies in their serum, phenotyped as McCoy (a−) at birth and, on retesting their red blood cells, 1 at 5 months and the 2nd at 11 months, both phenotyped as McCoy (a+). It is possible that this lack of expression of the McCoya antigen at birth represents a characteristic of an obligate heterozygote. However, on testing 50 random umbilical cord red blood cell samples with two potent McCoya antisera, only 1 was found to be negative. If the negative phenotpye were a characteristic of an obligate heterozygote at birth, then the neonatal incidence of the McCoy (a−) phenotype should have been significantly (p<10−4) higher in our random cord blood sample than observed (expected 11). The impaired expression of the McCoya antigen on the red blood cells of these 2 infants at birth, in conjunction with evidence of maternal alloimmunization, strongly suggests alloantibody‐induced antigenic suppression of the McCoya antigen as a probable cause.
Molecular Sieve Chromatography of Some Plasma Protein Products by High Performance Liquid Chromatography: Comparison of Quantitation by UV Absorption and Lowry Protein
Authors: Bertolini Maurice J.
Published: March 2009

Abstract. Commercially prepared fraction V products (5 and 25% albumin and 5% plasma protein fraction) and immune serum globulins were subjected to analytical molecular sieve chromatography by high performance liquid chromatography. Quantitation was performed by UV absorbance and by the Lowry protein assay. The proteins eluting in the void volume from the immune serum globulin samples yield equivalent results by UV absorbance and by Lowry assay. Proteins eluting in the void volume from fraction V products indicate approximately twice as much protein by UV absorbance when compared to Lowry protein assay.
Management of Immune‐Mediated and Paraproteinemic Diseases by Membrane Plasma Separation and Cascade Filtration
Authors: Valbonesi M. , Garelli S. , Montani F. , Cefis M. , Rossi U.
Published: March 2009

Abstract. Membrane plasma separation was utilized in the management of 19 patients with autoimmune, immune complex or paraproteinemic diseases. In these patients 34 procedures were carried out employing a new filter composed of polyvinyl alcohol hollow fibers with a nominal pore size of 0.4 μm. Cascade filtration was performed in 23 procedures using a second filter built up with ethylenevinylalcohol copolymer hollow fibers with a nominal pore size of 0.1–0.2 μm. Both filters employed showed a very good biocompatibility, and no untoward effect was observed. Using the differential sieving effect of cascade filtration it was possible to eliminate circulating immune complexes and immunoglobulin M without the loss of necessary plasma components.
Catabolic Half‐Lives and Antigenic Relationships of Native, Altered and Commercially Prepared Human Albumins in Rabbits
Authors: Anhorn C. , Sheldon S. , Laschinger C. , Naylor D. H.
Published: March 2009

Abstract. As determined in rabbits, the in vivo plasma survival, i.e. half‐life (t 1/2), of albumin produced by the cold‐ethanol procedure did not differ significantly from that of native albumin (p>0.5). Plasma survivals of reprocessed and heat‐ethanol produced albumins were significantly less than that of the native protein (0.02 <p< 0.05). Chemically‐modified albumin had a considerably reduced t 1/2 (p<0.001) and was the only albumin shown to be antigenically different from the native protein. Variations in t 1/2s of albumin preparations could not be attributed to differences in their polymer contents.
Serological Activity of Low Frequency Antigens of the MNSs System and Reappraisal of the Miltenberger Complex
Authors: Giles Carolyn M.
Published: March 2009

Abstract. Low frequency antigens of the MNSs system were tested for sensitivity to trypsin treatment which subdivided them into two distinct groups. A new interpretation of the Miltenberger complex based on determinants rather than type sera is proposed.
Elimination of Monomeric, Dimeric and Polymeric Human Albumin from the Plasma of Inbred Mice and Dissociation of Natural Albumin Polymers
Authors: Ferner P. , Anhorn C.A. , Laschinger C. , Naylor D. H.
Published: March 2009

Covalently linked albumin dimers and polymers had plasma half‐lives (t 1/2s) in mice which were not significantly different from that of native albumin. The elimination of a chemically treated albumin monomer was also not appreciably different from that of the native protein, but it was significantly faster than covalently linked albumin dimers and polymers. Essentially similar results were obtained when natural monomers, dimers and trimers were compared, although precise t 1/2 data may have been affected by the dissociation of natural polymers into appreciable amounts of monomer in vivo.
Plasma Exchange and Immune Complex Diseases: The Predictability of Immune Complexes Removal to Clinical Response
Authors: Valbonesi M. , Garelli S. , Montani F. , Manca F. , Cantarella S.
Published: March 2009

58 patients were treated by discontinuous flow plasma exchange because of an immune complex (IC) disease. 41 patients who had a high level of circulating IC prior to plasmapheresis showed both clinical and immunochemical evidence of improvement with plasma exchange. Only 2 patients with a high level of IC did not improve after therapy: these patients suffered from multiple sclerosis and idiopathic thrombocytopenic purpura, respectively. Prior to PE therapy the level of circulating IC in 15 patients was within the normal range, when measured according to Manca et al. In this group none of the patients worsened after therapy, whereas 8 patients showed an objective improvement. The positive correlation between IC removal and clinical results suggests that patients with a high level of circulating IC are most likely to benefit from apheretic treatment.
Factors Influencing Changes in pH during Storage of Platelet Concentrates at 20–24°C1,2
Authors: Moroff Gary , Friedman Andrea , Robkin‐Kline Linda
Published: March 2009

The magnitude of the pH change during platelet concentrate storage at 20–24°C in polyvinyl chloride containers is not determined solely by platelet count per cubic millimeter of plasma, since a wide variation in pH was observed with similar platelet concentrations. In modified platelet concentrates having lost through centrifugation 3–15% of total platelets and 61–92% of residual leukocytes, the pH was maintained at substantially higher levels than in the paired control platelet concentrates. Leukocyte levels do not appear to determine the magnitude of the pH fall. Continuous oxygen ulilization is needed if the pH is to be maintained near 7.0. However, oxygen tension per se is not the factor which influences the extent of pH change. It has been concluded that a specific platelet subpopulation comprising a small proportion of the total platelets in concentrates and having an enhanced capacity to form lactate may be responsible for a major part of the pH reduction which occurs during storage of many platelet concentrates.
Management of Immune‐Mediated and Paraproteinemic Diseases by Membrane Plasma Separation and Cascade Filtration
Authors: Valbonesi M. , Garelli S. , Montani F. , Cefis M. , Rossi U.
Published: March 2009

Membrane plasma separation was utilized in the management of 19 patients with autoimmune, immune complex or paraproteinemic diseases. In these patients 34 procedures were carried out employing a new filter composed of polyvinyl alcohol hollow fibers with a nominal pore size of 0.4 μm. Cascade filtration was performed in 23 procedures using a second filter built up with ethylenevinylalcohol copolymer hollow fibers with a nominal pore size of 0.1–0.2 μm. Both filters employed showed a very good biocompatibility, and no untoward effect was observed. Using the differential sieving effect of cascade filtration it was possible to eliminate circulating immune complexes and immunoglobulin M without the loss of necessary plasma components.
Proficiency in Testing for Hepatitis B Surface Antigen: Experience with Ausria II®1
Authors: Nath Nrapendra , Fang Chyang T. , Pielech Mitchell , Lawson William , Dodd Roger Y.
Published: March 2009

Eight hepatitis B surface antigen (HBsAg) proficiency panels were tested using a radioimmunoassay (RIA) test kit during 1978 and 1979 by routine processing laboratories within the 57 American Red Cross Blood Services Regions. The test results were analyzed and it was found that lot‐to‐lot variability in the reagents may result in a measurable loss of sensitivity of the test leading to an increased error rate. On the basis of these studies, it is recommended that the positive to negative (P/N) ratio for validation of the test be raised to at least 25.
Precipitin Reaction between Sda‐Active Human Tamm‐Horsfall Glycoprotein and Anti‐Sda Serum
Authors: Serufini‐Cessi F. , Conte R.
Published: March 2009

Abstract. Human Tamm‐Horsfall glycoprotein isolated from an Sd(a+) donor, with high Sda inhibitory activity, was labelled with 14C‐formaldehyde. The labelled glycoprotein was precipitated by anti‐Sda sera. A good correlation was found between the precipitating and agglutinating titers of different anti‐Sda sera.
Elimination of the Prozone Effect in the Antiglobulin Reaction by a Simple Modification
Authors: Salarna A. , Mueller‐Eckhardt C.
Published: March 2009

Abstract. A simple modification of the antiglobulin test (AGT) is described. It consists of one additional washing after antiglobulin incubation. This increases the sensitivity of the test and eliminates the prozone phenomenon in the direct as well as the indirect AGT.
False Reactions in Radioimmunoassay for Viral Hepatitis B Markers in Patients Suffering from Coagulation Disorders
Authors: Grangeat‐Keras L. , Lambert T. , Dubreuil P. , Briantais M.J. , Pillat J.
Published: March 2009

Abstract. Simultaneous occurrence of hepatitis B surface antigen (HBsAg) and antibodies to HBsAg (anti‐HBs) has been frequently described in hemophiliac patients. During our working procedures, this phenomenon was also observed. Interference of coagulation disorders with radioimmunoassay (RIA) used in the detection of viral hepatitis B markers was studied. It was shown that, in vivo or in vitro, correction of coagulation disorders led to the disappearance of false positive reactions and, thus, simultaneous presence of HBsAg and anti‐HBs never occurred. Confirmatory tests which are usually recommended to control RIA were reviewed and it was found that only a completed coagulation of serum by thrombin addition, before performing the test, gave specific results.
Production of Direct Agglutinins by the Combination of Incomplete Antibodies to Protein A‐Bearing Staphylococcus aureus Cells
Authors: Deary I.J. , Moore S. , Pepper D.S.
Published: March 2009

Abstract: A novel method is described for the production of saline‐reacting antibodies from ‘incomplete’ blood grouping sera. IgG anti‐D is combined to protein A‐bearing Staphylococcus aureus (SA), forming a polyvalent anti‐D assembly. The SA‐anti‐D has a high titre in saline and is stable on 4°C storage and lyophilisation. The material was successful in Rh(D) typing cells sensitised in vitro with various antibodies. Saline‐reacting SA‐anti‐Kell and SA‐anti‐c̄ were also produced.
Effects of Source Leukocyte Collection on the Immune System
Authors: Orsini F. , Fitzpatrick J. , Cohen E. , Khurana U. , Michielson C. , Chadha K.C. , Han T. , Ozer H.
Published: March 2009

This study was initiated in order to assess any immunologic effects that source leukocyte concentrate collection might have on double‐bag plasmapheresis donors. Previous studies have shown that surveillance parameters, such as T versus non‐T lymphocyte subpopulations, showed no abnormal values in donors with as many as 500 visits over a 12‐year period. The present study demonstrates that the frequency and the total number of leukocyte donations do not effect the lymphocyte subpopulations and functions observed. No significant changes were noted for specific and nonspecific stimulation, natural killer (NK) cell activity, lymphocyte surface markers and a variety of functional parameters. For example, the apheresis donors manifested no differences in NK cell activity, human leukocyte interferon production, IgG synthesis by B cells and percent suppression of both IgG synthesis and mixed lymphocyte cultures, when compared to non‐apheresed donors.
Normal Viability of Platelet Concentrates Obtained from CPDA‐1 Blood after Storage in CL‐3000 Blood Containers
Authors: Simon Toby L. , Murphy Scott
Published: March 2009

Platelet concentrates were obtained from blood anticoagulated with CPDA‐1 and their viability after storage in CL‐3000 containers for 72 h at 22°C and for 24 and 48 h at 4°C was studied using autologous reinfusion of 51Cr‐labeled platelets. Yield and t1/2 values after such storage were similar to those previously reported for other containers and anticoagulants. Survival of platelets stored at 22°C for 3 days was essentially normal with the expected yields for that duration of storage. As has been described for other containers, adenine has no adverse or beneficial effect on platelet viability when assessed with these methods.
A New Therapeutic Antibody Removal Method Using Antigen Positive Red Cells: Application to M‐Incompatible Pregnant Women
Authors: Yoshida Hisahiro , Yoshida Yataro , Tatsumi Kenichi , Asoh Takeshi
Published: March 2009

A study was made for the therapeutic removal of red cell antibodies using antigen positive red cells. With preliminary experiments in vitro, the criteria for the optimal absorption of anti‐M with O MM red cells were estimated to be the following: an incubation time of 10min, an incubation temperature of 0–4°C (in an ice‐water bath), and a red cell‐to‐plasma ratio of 1:2.5.

On the basis of these in vitro experiments, the following procedure was designed. One unit (about 90 ml) of O MM packed red cells was added to a bag containing 250 ml of patient's plasma, the mixture was incubated in an ice‐water bath for 10 min. Following centrifugation, autologous plasma from which the antibodies were removed was reinfused. After absorption the titer of anti‐M fell from 512 to 4. No variation in the level of other plasma components was detected and no hemolysis was seen. After 2.5 *** of patient's plasma was treated using the above described method, the antibody titer usually decreased one log2 unit.

When this method was compared with exchange plasmapheresis, no significant abnormalities in the immunoglobulin and hemostatic factors could be detected, but a moderate decrease in platelet count was seen after plasmapheresis. Thus, it appears that our method provides a rational therapeutic modality for specifically removing antibodies.
Rapid Phenotyping of C3 by Immunofixation on Cellulose Acetate Strips
Authors: Germenis A. , Babionitakis A. , Fertakis A.
Published: March 2009

Determination of the genetically controlled variants of the C3 system has been accomplished by electrophoresis on cellulose acetate membrane followed by immunofixation with a specific anti‐C3 antiserum. The results have been confirmed with the use of reference sera and compared with those obtained from conventional high voltage agarose gel electrophoresis. The interference of some electrophoretic variables on the technique has been investigated. The method is rapid, accurate, reliable and gives good separation of at least the three common C3 phenotypes.
Intravenous Immune Serum Globulin in Immunodeficiency
Authors: Cunningham‐Rundles Charlotte
Published: March 2009

This study was undertaken to compare intravenous gamma‐globulin (IVGG) for 1 year in patients with primary immune defects who had previously been treated with intramuscular gamma‐globulin (IMGG). Forty‐three patients were available for analysis. After administration of 300 mg/kg of IVGG every 3 weeks, blood immunoglobulin levels increased to within 1 SD of normal for the patient's ages. In 5 patients, this dosage regimen did not raise the immunoglobulin levels satisfactorily, although they were still better than those achieved with IMGG. In 18 patients, treatment benefits became apparent after 5–6 months of treatment. Substantial improvement was observed in 70% of these patients. Sixteen reactions occurred with 638 infusions, an incidence rate of 2.5%. If 2 patients with an IgM macroglobulin and 1 patient with an anti‐IgA antibody were excluded, the reaction incidence would probably have been 1%.
Development of CMV Antibody Tests and Their Clinical Evaluation
Authors: Wielaard F. , Scherders J. , Dagelinckx C. , Hooijmans A. , Smit‐Siebinga C. T. , Welle F.
Published: March 2009

Abstract. An anti‐CMV ELISA was developed in which monoclonal antibodies to immobilize the CMV antigens are used. The test was compared with the IHA test and there was 98.6% agreement. Using the CF test as a referee, sensitivity and specificity of the ELISA were calculated to be 98.6 and 99.0%, respectively. The seroconversion rate among Dutch donors was estimated from the age‐related prevalence of anti‐CMV antibodies to be about 0.4%. Specific anti‐CMV IgM was searched for in a large donor population using an antibody‐capture IgM ELISA; 1 of 600 donor sera was found positive. This makes it unlikely that screening for IgM anti‐CMV will effectively prevent posttransfusion CMV infections.

In order to meet the demand for simpler tests, a one‐step ELISA was recently developed, based on the inhibition principle. Preliminary studies with this test indicate a high sensitivity and specificity.
Application of Robotics in Blood Banking1,2
Authors: Friedman Leonard I. , Severns Matthew L.
Published: March 2009

Abstract. To demonstrate the feasibility of using robots in blood banking applications several prototype systems were developed. Activities associated with sample testing and component preparation were examined. In one project, a general‐purpose laboratory robot (Zymate Laboratory Automation System, Zymark, Inc., Hopkinton, Mass.) was configured to prepare samples for microplate‐based ABO/Rh testing. In a second project, this same robot was configured to carry out specific steps in evaluating bar‐coded labels, as part of a quality control procedure.

A fluid‐handling robot (Sampler 505, Tecan AG, Hombrechtikon, Switzerland) was used to prepare the dilution of serum samples for the evaluation of an anti‐HTLV‐III test kit. It was then configured to aspirate, dilute and dispense samples for anti‐HTLV‐III and HBsAg testing. This robotic system is now in field trial.

The use of large industrial robots for automating component production was also considered. The key element in this design was the development of a fixture that would hold the blood bag set during the balancing, centrifugation and expressing steps. However, a fixture which was capable of performing these operations and that was equivalent in size and adaptable to a standard centrifuge bucket could not be fabricated.
Immunologic Disorders: The Regulation of Humoral Immunity
Authors: Wedgwood Ralph J. , Ochs Hans D.
Published: March 2009

Abstract. The modulation of humoral immunity by complement and complement receptors is presented, as well as the importance of the X chromosome in immune response. Gene rearrangement and ontogeny of the immunoglobulin isotypes are discussed with regard to humoral immunodeficiency diseases.
Children with AIDS
Authors: Pahwa Savita
Published: March 2009

Abstract. The etiologic agent for acquired immune deficiency syndrome (AIDS) has been identified as human T lymphotropic virus type III (HTLV‐III). Its distinguishing characteristics and putative receptor, the T4 molecule, are discussed. The Western Blot method has been applied to assess the presence of antibody to HTLV‐III in patients as a measure of infection with HTLV‐III. The definition of AIDS in children, based on the Centers for Disease Control surveillance criteria, is explored and expanded. Clinical and laboratory manifestations of pediatric AIDS are explained in the context of ‘HTLV‐III infection’ through an analysis of prospective and retrospective serologic studies. Transmission of the disease is explored through parental history of infected children. Finally, the rationale for antibody replacement therapy is set forth.
Major Histocompatibility Complex Class I to III Allotypes in Patients with AIDS‐Related Complex/Walter‐Reed 5, Disseminated Kaposi's Sarcoma and in Normal Controls
Authors: Plum Georg , Siebel Eberhard , Bendick Christoph , Rasokat Heinrich , Braun Margot , Krickeberg Holger , Mauff Gottfried ,   ARC‐IVIG Study Group Cologne/ Berne
Published: March 2009

Abstract. In HIV‐infected patients major histocompatibility complex (MHC) class I and II (= HLA‐A, B, C, DR) association has been controversial. Of the MHC class III coded complement components C2, BF, C4A/C4B especially C4 allotypes appear of major immunogenetic relevance for their potential differences in virus neutralizing potency and immune complex formation. In the present study 29 patients with AIDS‐related complex and Walter‐Reed 5 (ARC/WR5), 35 patients with disseminated Kaposi's sarcoma (KS), and 160 HIV‐negative control individuals were compared for MHC class I to III allotypes. Diagnosis of ARC and KS (WR criteria) was done by clinical and laboratory parameters, MHC testing, by standard procedures. An increase in frequency (p≤0.05) was observed between ARC/WR5 patients and controls for HLA‐B35/CW4, DRW14, a decrease for B16, CW6/DR7. However, values were not significant if corrected for the number of tested antigens. No significant differences were seen between KS and ARC patients or controls for class III allotypes, nor for previously reported associations, e.g. for B8, DR2, DR3, and especially DR5, including the DR5 splits DRW11, 12. The results indicate the lack of a strong MHC association with the investigated antigens in West German Caucasoids, and support the hypothesis of ethnic dependence of HIV‐related diseases. The HLA‐B35/CW4 increase, also associated with the duplicated C4 A*3 A*2 and the silent C4B*Q0, was more pronounced in ARC patients with progression to AIDS‐OI. The increased frequency of C4B*Q0 alleles in these patients was thought to be secondary to a hypothetical increase in ‘converted’ and dysregulated C4 genes not seen to be associated in this study.
Hepatitis A Virus Transmission by Blood Products in the United States
Authors: Mosley James W. , Nowicki Marek J. , Kasper Carol K. , Operskalski Eva A. , Donegan Elizabeth , Aledort Louis M. , Hilgartner Margaret W. ,   The Transfusion Safety Study Group
Published: March 2009

To address the question of HAV prevalence and seroconversion in relation to clotting factor concentrates, we assayed an early serum for 339 hemophiliacs followed every 6 months by the Transfusion Safety Study in the period from mid‐1985 until mid‐1992. We found 58.4% positive around entry, with an age‐specific prevalence that did not vary with age. In comparison to rates for anti‐HIV‐negative blood donors, they were significantly higher. Based on testing of subsequent sera, 11 hemophiliacs (7.8% of 141 susceptibles) changed their anti‐HAV status from negative to positive. In 9 instances, positivity immediately followed the first dose of intravenous immune globulin. A possible seroconversion followed treatment with blood components, and a possible seroconversion followed intermediate‐purity, solvent/detergent(SD)‐treated factor VIII concentrate. Neither of these 2 patients was anti‐HAV IgM positive, so that passively transferred antibody is possible. The high prevalence among hemophiliacs at entry must be further investigated by determining the expected background rate in a US population similar to the hemophilia patients, and by comparing anti‐HAV prevalence associated with SD‐treated and heat‐inactivated concentrates.
Sequence Variability of Hepatitis A Virus and Factor VIII Associated Hepatitis A Infections in Hemophilia Patients in Europe: An Update
Authors: Robertson Betty H. , Friedberg Devorah , Shouval Daniel , Normann Andrea , Graff Judith , Flehmig Bertram
Published: March 2009

Outbreaks and sporadic cases of hepatitis A have been observed in 4 European countries in hemophilia patients receiving factor VIII preparations. PCR amplification of potential hepatitis A virus (HAV) nucleic acid present in plasma pools, purified factor VIII and acute‐phase sera from infected individuals has been performed and the nucleic acid sequence determined for those samples that resulted in a positive PCR product. HAV sequences were detected in the serum of 2 German patients, but not in the factor VIII lots administered to these individuals. Screening of plasma pools and the corresponding 5 lots of factor VIII associated with the outbreak in Ireland did not reveal any HAV sequences. In contrast, a study of samples from Italy detected HAV sequences in 5 of 12 lots and in 2 hemophilia patients who developed hepatitis A. These data suggest that implicated factor VIII preparations might have been involved in the outbreaks of HAV infection among Italian hemophiliacs. However, no molecular evidence was obtained for a similar association in Germany or Ireland. The preliminary data from these two investigations must be verified by animal inoculation studies and supported by epidemiologic analysis.
Distribution of Hepatitis A Antibody over a Process for the Preparation of a High‐Purity Factor VIII Concentrate
Authors: Hart H. , Jones A. , McIntosh R.V. , Cuthbertson B. , Cubie H.
Published: March 2009

The levels of hepatitis A antibody were studied in factor VIII products manufactured by the Scottish National Blood Transfusion Service. It was found that the current high‐purity factor VIII product has no detectable hepatitis A antibody, whereas the superseded intermediate‐purity product had significant antibody levels (6,700 mIU/ml). Although the finished high‐potency factor VIII product has no detectable hepatitis A antibody, significant levels of antibody are found in the early stages of product manufacture. This antibody may offer some protection against hepatitis A contamination present in very occasional plasma donations. Antibody to parvovirus B19 is also present at intermediate stages in the manufacture of high‐potency factor VIII, but its significance is not known. Hepatitis A antibody levels were also measured in normal immunoglobulin. These data indicate that antibody levels in the plasma pools used to manufacture factor VIII are falling.
Detection of Hepatitis A Virus in a Factor VIII Preparation by Antigen Capture/PCR
Authors: Normann Andrea , Graff Judith , Flehmig Bertram
Published: March 2009

The antigen capture/PCR (AC/PCR) has been applied in the analysis of various factor VIII preparations, which were suspected to be contaminated with hepatitis A virus (HAV). AC/PCR involves capturing the antigen, i.e. the intact virus particles, by binding to the HAV monoclonal antibody mAb 7e7, reverse transcription of the viral RNA and amplification of the cDNA with HAV‐specific primer pairs. The PCR analysis of one factor VIII concentrate yielded an HAV‐specific DNA product, which could be confirmed by Southern blot analysis. The HAV strain recovered by AC/PCR from this factor VIII concentrate could be classified into genotype III after solid‐phase sequencing of the product and comparison with the consensus sequences for the known HAV genotypes. Analysis of the sera from 3 haemophiliacs treated with this batch has not resulted in a reliable product. However, the results obtained indicate that HAV can be detected in purified factor VIII preparations by AC/PCR.
Removal and Inactivation of Enveloped and Non‐Enveloped Viruses during the Purification of a High‐Purity Factor IX by Metal Chelate Affinity Chromatography
Authors: Roberts Peter L. , Walker Chris P. , Feldman Peter A.
Published: March 2009

Virus reduction during the copper chelate affinity chromatography stage used during the purification of a new high‐purity factor IX (BPLs 9MC) has been investigated. Virus reduction for the enveloped virus Sindbis was 6.5 log, a value which included approximately 2 log of inactivation due to the use of an acidic wash buffer (pH 4.4) during chromatography. In the case of the non‐enveloped hepatitis‐A‐like poliovirus, which is acid‐resistant, the virus reduction value was 4.0 log and was exclusively due to physical virus removal during the chromatographic process.
Removal and Inactivation of Hepatitis A Virus (HAV) during Processing of Factor VIII Concentrates
Authors: Hamman J. , Zou J. , Horowitz B.
Published: March 2009

Although hepatitis A virus (HAV) transmission by single donor blood products is a rare event, recent reports have raised concerns about the potential for HAV transmission by pooled plasma products. In this study the removal and inactivation of HAV during production of a factor VIII concentrate, which included solvent‐detergent treatment and anion‐exchange chromatography steps, was assessed at 5 distinct stages of production using infectivity and, in some cases, isotopic assays. The results of these ‘spiking’ studies revealed an overall HAV clearance of >8 log with appreciable contributions from antibody neutralisation, cryoprecipitation, lyophilisation and possibly chromatography, but not filtration. In addition it was shown that neither solvent detergent treatment or ion‐exchange chromatography reversed or impeded the neutralisation of HAV by antibody.
Virology of the Hepatitis A Epidemic in Italy
Authors: Purcell Robert H. , Mannucci Pier M. , Gdovin Susan , Gringer Alessandro , Colombo Massimo , Melec Alfonso , Schinaia Nicola , Ciavarella Nicola , Emerson Suzanne U.
Published: March 2009

We studied the virologic aspects of a hepatitis A epidemic that occurred among hemophilia patients in Italy between 1989 and 1992. Twelve lots of factor VIII concentrate manufactured by the solvent‐detergent chromatographic technique and suspected of contamination by the hepatitis A virus (HAV) were analyzed by a two‐step, nested polymerase chain reaction (PCR) procedure. PCR was applied to 1‐ml samples of factor VIII concentrate and 100‐μl serial serum samples available from 2 patients. Particular care was taken to rule out the possibility of false‐positive results during analysis. Results demonstrated PCR amplification of the 3′‐region of the VP3 gene in 5 of the 12 implicated lots of factor VIII and in the serial serum samples of both patients. PCR amplification also revealed that the gene sequences detected in patients' sera were identical to the sequences detected in the product they had received. In all, 3 VP3 sequences (found to be 96–99% identical) were amplified. Further characterization of the HAV found in the factor VIII concentrate and the patients' sera was attempted by PCR amplification of the VP1/2A region. Successful amplification of this region was achieved in the serum of only 1 patient and in the concentrate he received. This fourth amplified sequence was identical in both serum and factor VIII concentrate. Attempts to transmit hepatitis A from the contaminated lots to 3 chimpanzees resulted in no signs of infection after 10 months of observation. Based on the Italian experience, persons with severe hemophilia who receive large‐pool concentrate are at potential risk for HAV infection and should be vaccinated against HAV or use an alternative to solvent‐detergent‐prepared concentrate. Since June 1992, no cases of HAV have been reported in Italian hemophiliacs, most of whom have been vaccinated.