bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of the Reactions of the Rh‐Related Murine Monoclonal Antibodies BS58 and R6A
Authors: Sonneborn H.‐H. , Ernst M. , Tills D. , Lomas C. G. , Gorick B. D. , Hughes‐Jones N. C.
Published: March 2009

The murine monoclonal antibodies BS58 and R6A are known to recognize epitopes related to the human Rh system: neither antibody reacts with Rhnull cells and the BS58 antigen is not expressed by‐D‐or ·D· cells. It is shown here that the numbers of BS58 and R6A antigen sites vary with Rh phenotype. Both epitopes are well represented on cells of the CDe/CDe, CDe/cDE and CDe/cde phenotypes; BS58 sites are markedly reduced on cde/cde and cDE/cde and are only just detectable on cDE/cDE cells when compared with R6A sites. The number of R6A sites per red cell ranged between 20,000 and 150,000. The evidence indicates that the BS58 epitope is not on the polypeptides carrying D or R6A, nor is it uniquely on one of the polypeptides carrying either C, c, E or e. It is suggested that the BS58 epitope is either common to all the CcED polypeptides or that it is present on a polypeptide which has not yet been identified biochemically.
Serological and Immunochemical Specificity of a Human Autoanti‐Gerbich‐Like Antibody
Authors: Pookle J. , Reid M. E. , Banks J. , Liew Y. W. , Addy J. , Longster G.
Published: March 2009

Abstract. An 85‐year‐old male with cardiac failure secondary to anaemia had an apparent anti‐Ge2 (Ge = Gerbich) in his serum which did not agglutinate his own red cells even though they were Ge‐positive in tests with alloanti‐Ge. The direct antiglobulin test was negative; however, an antibody with apparent anti‐Ge2 specificity was eluted from his red cells. The patient's autoantibody was shown in immunoblotting experiments to react with an antigenic determinant on (3‐sialoglycoprotein. This case illustrates that an autoanti‐Ge can masquerade as an alloantibody, thereby complicating antibody identification, and implies that the immunochemical specificity of autoanti‐Ge2 is different from that of alloanti‐Ge2.
Electron Microscopy of Red Blood Cells Altered by Auto‐Inimunity‐Inducing Drugs
Authors: Rahamim Ezra , Kahane Amnon , Sharon Rafael
Published: March 2009

Abstract. The fact that levodopa, a drug used for the treatment of Parkinson's disease induces a direct Coombs‐positive reaction in about 15% of treated patients, indicating the presence of auto‐antibodies against the patient's red blood cells (RBCs), is well known. Another known fact is that only 1% of those patients do actually develop auto‐immune haemolytic anaemia. In this paper, we describe our findings utilizing the direct ELISA, a method for measuring the presence of IgM and IgG auto‐antibodies on the patients' red blood cells (RBCs), as well as an indirect ELISA, testing the presence of antibodies in their serum. The tests were performed on 8 patients, 7 of whom had been receiving the drug for long periods of time. Our results show that the serum antibodies precede the bound antibodies in a significant fashion and that the severity of the anaemia is directly related to the amount of auto‐antibodies. Since auto‐antibodies do not seem to be directed against the drug itself, some modification of the RBC must occur. For this reason, we attempted to determine whether patients receiving the drug show any morphological changes of their RBCs which could be determined by scanning electron microscopy (SEM) and whether these changes could be related to direct and indirect ELISA. In most levodopa‐treated patients, a small number of echinocytes could be observed by SEM, while in 1 patient who suffered a severe anaemic phase, a very high percentage of the RBCs were echinocytes and spherocytes. In vitro treatment of normal human RBCs showed similar forms of transformed RBCs, possibly explaining the reason for the auto‐immune response caused in patients with renal failure undergoing extracorporeal haemodialysis with formaldehyde‐resterilizable membranes.
Monoclonal Antibody to Blood Group Glycosyltransferases, Produced by Hybrids Constructed with Epstein‐Barr‐Virus‐Transformed B Lymphocytes from a Patient with ABO‐Incompatible Bone Marrow Transplant and Mouse Myeloma Cells
Authors: Kominato Yoshihiko , Fujikura Takashi , Shimada Ichirou , Takizawa Hisao , Hayashi Kyoko , Mori Takao , Matsuda Tamotsu
Published: March 2009

Abstract. A patient with chronic myeloid leukemia secreted an antibody to blood group glycosyltransferases after ABO‐incompatible bone marrow transplantation (B recipient/O donor). Peripheral B lymphocytes from the recipient were transformed with Epstein‐Barr virus, and then fused by polyethylene glycol with mouse myeloma cell line P3‐X63/Ag8.653. After the cloning of the hybridoma cells, a cell line which produced human IgM antibody to blood group glycosyltransferases was established. The antibody completely neutralized B transferase activity at low concentration, while a larger amount of immunoglobulins was required to neutralize A transferase activity.
The Influence of Cryopreservation on Activity and Surface Markers of Lymphokine‐Activated Killer Cells
Authors: Letellier C. , Rameliarison L. , Fizet D. , Ferrer A.M. , Vezon G.
Published: March 2009

Ficoll‐separated and monocyte‐depleted mononuclear cells isolated from normal leukapheresis products were cryopreserved. These cells were incubated with or without 1,000 U/ml of recombinant interleukin‐2 (rIL‐2) for 4 days, and their lymphokine‐activated killer (LAK) and natural killer (NK) activities were measured. IL‐2 activation induced a significant increase in the expression of the CD25 antigen. There was no change in CD2, CD3, CD4, CD8, CD16, CD56 and CD57 cell marker expression. Cryopreservation did not induce any change in the membrane antigen expression and in the lymphocyte subsets. The NK activity was well preserved and the decrease of LAK activity of IL‐2‐activated cells after cryopreservation was not significant. In contrast, cells activated before cryopreservation had a significantly lower cytotoxic activity and the number of cells expressing the IL‐2 receptor was also significantly reduced. However, the decrease of CD56 expression was not significant. CD25 expression seemed to be proportional to the LAK activity of the cells. This study demonstrated that cryopreserved lymphocytes, after 4 days of culture with rIL‐2, could be as active and could express the CD25 and CD56 cell surface markers in the same manner as fresh LAK cells.
HLA Typing and Lymphocytotoxic Crossmatches using Conventional Techniques or Immunobeads1
Authors: Mueller‐Eckhardt G. , Kölzow S. , Conrath K. , Hoffmann O.
Published: March 2009

In a prospectively designed comparative study, HLA class I and class II typing was performed after B/T lymphocyte separation by nylon wool (NIH technique) or using immunomagnetic beads (IMB technique). Samples from 70 healthy individuals (group A) and from 47 patients with potential typing problems (group B) were examined. Complete typing failures occurred more frequently with NIH than with IMB. Except for class I in group A, the rate of correct decisions for antigen assignment was significantly higher with IMB. For class II serum reactions, specificity, predictive value and efficiency were superior for IMB, while sensitivity appeared higher for NIH. This can possibly be explained by a prozone effect which was observed in 9 of 36 titration experiments only with the IMB technique. Correlation of both methods in 104 crossmatches prior to kidney transplantation was poor using peripheral blood lymphocytes, but excellent with spleen cells. In addition to the important advantages of the IMB technique for potential problem cases, especially for class II, typing time is reduced to one third, which is of particular clinical importance in kidney transplantation. However, lymphocytotoxic crossmatches prior to transplantation, if performed by IMB, should be carried out using different serum dilutions or with donor spleen cells instead of peripheral blood lymphocytes.
Posttransfusion Fulminant Hepatitis B Associated with Precore‐Defective HBV Mutants
Authors: Kojima M. , Shimizu M. , Tsuchimochi T. , Koyasu M. , Tanaka S. , Iizuka H. , Tanaka T. , Okatnoto H. , Tsuda F. , Miyakawa Y. , Mayumi M.
Published: March 2009

Abstract. Fulminant hepatitis B developed in 8 recipients of blood units without detectable hepatitis B surface antigen on routine screening. All 124 hepatitis B virus (HBV) DNA clones propagated from their sera possessed defects in the precore region. A point mutation from guanine to adenine at nucleotide 83, converting codon 28 for tryptophan (TGG) to a stop codon (TAG), was the commonest, and it was found in all 113 clones from 7 cases. The remaining case displayed 1 clone with this point mutation and 10 clones with an insertion of 2 base pairs after nucleotide 26. Antibody to hepatitis B core antigen (anti‐HBc) was detected in a high titer in 1 of 10 pilot plasma samples of blood units transfused to this case. HBV DNA clones propagated from it exhibited the same precore‐region defects as those from the recipient. On the basis of these results HBV mutants, defective in the precore region, would appear to be responsible for posttransfusion fulminant hepatitis B, and the exclusion of blood units with high‐titered anti‐HBc would be efficacious in preventing it.
Elution of Anti‐Zwa (‐PlA1) from Autologous Platelets after Normalization of Platelet Count in Post‐Transfusion Purpura
Authors: Taaning Ellen , Skov Fleming
Published: March 2009

Abstract. In two patients suffering from post‐transfusion purpura, anti‐Zwa was demonstrated in ether eluates from autologous platelets harvested during the remission phase and after normalization of the platelet count. In both patients, platelet‐associated immunoglobulin (PAIg) was demonstrated during the acute phase. PAIg disappeared concomitantly with recovery and, thus, seems to be associated with the thrombocytopenia. These data support the assumption that immunocomplexes are adsorbed nonspecifically to the platelets and cause destruction of autologous platelets lacking the corresponding antigen. An association between the IgG3 subclass of anti‐Zwa antibodies and the destruction of autologous platelets was also seen.
Inactivation of Lipid‐Enveloped Viruses in Proteins by Caprylate
Authors: Lundblad J. L. , Seng R. L.
Published: March 2009

Abstract. The use of caprylate for the inactivation of lipid‐enveloped viruses in biologically active proteins both plasma derived and produced by cell culture was evaluated. Viruses consisted of herpes simplex virus type I, vesicular stomatitis virus, vaccinia virus, and Sindbis virus. Utilizing the dissociation reaction and varying the concentration of the ionized form of caprylate, a specific amount of the nonionized form of caprylate was maintained over a wide pH range. Virus‐spiked protein solutions contacted with caprylate provide rapid virus inactivation under a variety of conditions while maintaining the integrity of the respective protein or activity. With the exception of coagulation factor AHF, protein and biological activity yield were essentially quantitative. Caprylate is removed after treatment by size exclusion chromatography or anion/cation exchange adsorption of the protein, followed by buffer wash.
Evaluation of Bovine Serum Albumin Preparations and Ficoll Addition in Capillary Tube Hemagglutination
Authors: King M.‐J. , Crawford M.N. , Reid M.E.
Published: March 2009

Abstract. Eighteen commercial bovine serum albumin (BSA) preparations were evaluated for their efficacy in hemagglutination enhancement by the capillary tube method. Their physicochemical variables (pH, conductivity, free fatty acids, total protein, and polymer content) were quantified. The results showed that a BSA preparation which consistently performed well in hemagglutination enhancement contained some albumin polymer and had a low free fatty acid content. We also found that the addition of Ficoll (final concentration of 1.24%) could convert BSA preparations producing mediocre hemagglutination enhancement into good enhancers, provided that these preparations did not have high free fatty acid content. Hence, the addition of Ficoll can obviate the requirement for selecting a polymer‐enhanced albumin for use in capillary tube tests.
Alloantiserum Recognizing a DQw2 Split Which Is Associated with DR3
Authors: Flesch Brigitte , Neppert Jürgen , Ziegler Petra , Achtert Gudrun
Published: March 2009

Abstract. A typing serum MUE 38539 II, was found to recognize a DR3‐associated split of DQw2. In cytotoxicity tests, MUE 38539 II yielded positive test results with B lymphocytes but not with monocytes of DR3‐positive cell donors. This was in contrast to other typing reagents for DR3 that react with B lymphocytes as well as monocytes. Lymphocytotoxicity tests using MUE 38539 II were negative with DR7‐ and DQw2‐positive cells. The assumption that the serum recognizes a DR3‐associated split of DQw2, and not DR3 itself, was confirmed by the lack of reactivity with a DQw4‐ and DR3‐positive lymphoblastoid cell line (RSH). The assumption was also corroborated using reagents from a family in which DR3 and DQw2 were not found in the usually described linkage. In two lines, DR3 was associated with DQw‐ (2707 and 2710), and in the cell line 2704, DQw2 was associated with DRw‐. The serum MUE 38539 II was exclusively cytotoxic with lymphoblastoid cell lines from those family members who were positive for DQw2, independently of the DR3 antigens of the cells.
Autoimmune Haemolytic Anaemia due to Anti‐Phospholipid Antibodies
Authors: Arvieux J. , Schweizer B. , Roussel B. , Colomb M.G.
Published: March 2009

Abstract. The clinical course and laboratory findings of a patient with autoimmune haemolytic anaemia due to anti‐phospholipid antibodies and secondary to a systemic lupus erythematosus‐like syndrome are described. IgG and IgM with anti‐phospholipid activity coexisted in both sera and acid eluates prepared from the patient's red cells, as demonstrated by ELISA using a panel of anionic and neutral phospholipids. The anti‐erythrocyte binding activity of eluted autoantibodies, determined by a radioactive antiglobulin technique, was totally inhibited by absorption with phospholipid micelles. The patient's serum also contained an IgM warm haemolysin detectable with enzyme‐treated cells only, as well as anti‐C, ‐E and ‐S alloantibodies.
Chromatographic Preparation of a Therapeutic Highly Purified von Willebrand Factor Concentrate from Human Cryoprecipitate
Authors: Burnouf‐Radosevich M. , Burnouf T.
Published: March 2009

A therapeutic highly purified von Willebrand factor (vWF) concentrate has been prepared from cryoprecipitate by a three‐step chromatographic procedure. After solvent/detergent treatment to inactivate viruses, the cryoprecipitate solution was chromatographed on DEAE‐fractogel TSK 650 M to separate vWF from most cryoprecipitate proteins, including factor VIII (FVIII) and fibrinogen. A second DEAE‐fractogel TSK 650 M was then performed to further purify vWF and to allow concentrating it to over 100 U ristocetin cofactor activity/ml. The last step on immobilized gelatin removed fibronectin and increased the purity of vWF. vWF was recovered with about 18 and 40% yield in antigen and collagen‐binding (CB) activity, respectively, from cryoprecipitate. vWF was obtained in an essentially pure state corresponding to a purification factor of over 10,000‐fold from plasma. Immunonephelometric and SDS‐PAGE analyses of the concentrate did not reveal any detectable cryoprotein contaminants, especially fibrinogen, fibronectin, immunoglobulins and albumin. The content in intermediate‐ and high‐molecular‐weight multimers in the concentrate was similar or higher than that of plasma, as the ion‐exchanger selectively favored the binding and concentration of the larger multimeric forms while reducing the amount of the smaller forms with abnormal structure and low activity. Other characteristics of the concentrate included a CB activity to antigen ratio of 1.69 and a high capacity (86%) to correct platelet adhesion in a perfusion system. Clinical use of this standardized vWF concentrate has been shown to be efficacious in the treatment of vWF patients.
Note on the Measurement of Alanine Aminotransferase in the Screening of Blood Donors
Authors: Dorizzi Romolo Marco , Schiavon Renzo , Disperati Alberto , Chiavolini Paolo , Capuzzo Enrico
Published: March 2009

It should be remembered that nothing in nature stands alone; but that every art and science has relation to some other art or science, and that it requires a knowledge of these others, as far as this connection takes place, to enable us to become perfect in that which engages our particular attention. John Hunter [1]
Second‐Generation Anti‐HCV Tests Predict Infectivity
Authors: Larsen Jonn , Skaug Kjell , Maeland Arild
Published: March 2009

Twenty‐four blood donors found positive for the first‐generation hepatitis C antibody (anti‐HCV) test (Ortho EIA‐I) and 88 of their recipients over the period from 1972 to 1990 were retrospectively investigated with different first‐and second‐generation anti‐HCV tests. The aim of the study was to identify the infective donors and to evaluate the tests. Seven donors, who probably were infective carriers of HCV, were also second‐generation test (EIA‐II) positive, compared to only 3 out of 17 noninfective donors. Among the infected recipients, 14 out of 29 (48%) were positive for the second‐generation test only. The second‐generation test identified the infective donors in our study and was more sensitive than the first‐generation test. We therefore recommend that blood donors are screened with EIA‐II. Positive test results should be confirmed by the recombinant immunoblot assay (RIBA‐II), and persons with positive or not conclusive RIBA‐II should not be accepted as blood donors.
Prevalence and Classification of Anemia in Elective Orthopedic Surgery Patients: Implications for Blood Conservation Programs
Authors: Goodnough Lawrence Tim , Vizmeg Karl , Sobecks Ronald , Schwarz Adam , Soegiarso Wida
Published: March 2009

We audited 281 consecutive orthopedic patients scheduled for surgery for whom blood type/cross‐matching was requested over a 6‐month period. One hundred and sixty‐two patients predonated autologous blood at University Hospitals of Cleveland, and 34 (21%) of these were anemic [hematocrit (Hct) ≤39%] at initial donation. Twelve (35%) of these 34 anemic autologous blood donors subsequently received homologous blood. In contrast, 18 (15%) of 128 nonanemic autologous blood donors received homologous blood (p = 0.05). In 119 patients who did not donate autologous blood, 39 (33%) were anemic at admission. Of these, 22 (56%) received homologous blood. In the 80 remaining nonanemic patients, 33 (41%) received homologous blood (p = 0.119). Analysis of discharge Hct indicates that 31 (12%) of 263 evaluable patients were possibly transfused inappropriately. The anemias of a cohort of 30 autologous donors were analyzed: 5 had rheumatoid arthritis without iron deficiency. Nine (30%) others had evidence of iron deficiency. Sixteen (53%) had an unclassified anemia of chronic disease. We conclude: (1) the high rates of homologous blood exposure indicate a need for innovative blood conservation strategies in anemic autologous blood donors; (2) the prevalence of anemia and the high rates of homologous blood exposure in anemic patients who did not donate autologous blood demonstrate a need for early recognition and treatment in order to procure autologous blood and reduce homologous blood exposure; (3) the presence of inappropriate autologous and homologous transfusions demonstrates a need for more effective physician education programs that emphasize ‘no blood transfusion’ as an alternative to enhance blood conservation effectiveness.
Elevated Platelet‐Associated IgG in PIA1‐Negative Mothers following Sensitization to the PIA1 Antigen during Pregnancy
Authors: Kickler Thomas S.
Published: March 2009

Between 1984 and 1990, we studied 25 infants with neonatal alloimmune thrombocytopenia caused by alloimmunization to the PIA1 alloantigen. We investigated whether mothers of these infants developed elevated platelet‐associated immunoglobulin (PAIgG) in addition to anti‐PIA1. Eight of the women were found to have PAIgG which persistet at least 7–10 days postdelivery. Eluates prepared from six of the women's platelets reacted with PIA1‐positive and PIA1‐negative donor platelets, and platelets from donors with Glanzmann's thrombasthenia. None of the women with elevated PAIgG had thrombocytopenia, eclampsia, or infections. Two women were found to have autereactive antibodies present in plasma.

These results indicate that elevated PAIgG may be found in women immunized to the PIA1 antigen. Some women may also have autoreactive antiplatelet antibodies in their plasma. These findings may lead to confusing serologic findings in evaluating the cause of neonatal alloimmune thrombocytopenia.
Potassium Loss and Cellular Dehydration of Stored Erythrocytes following Incubation in Autologous Plasma: Role of the KCI Cotransport System
Authors: Olivieri Oliviero , de Franceschi Lucia , de Gironcoli Marzia , Girelli Domenico , Corrocher Roberto
Published: March 2009

We studied the regulation of cell volume and cation content in erythrocytes stored at 4°C under blood bank conditions for various lengths of time and subsequently incubated in autologous plasma at 37°C for 4 or 24 h. Cell swelled during storage at 4°C whereas marked K+loss and cell shrinkage were observed when erythrocytes were incubated at 37°C in autologous plasma. The cell shrinkage was inhibited only by the K+Cl‐cotransport‐specific inhibitor, [(dihydroindenyl)oxy] alkanoic acid, and not by other specific inhibitors of cation transport systems such as ouabain (Na+‐K+ATPase pump), bumetanide (Na+‐K+‐Cl‐cotransport) or carbocyanine (Ca++‐activated K+channel). Acidification and swelling of the erythrocytes are well known to be able to activate the K+Cl‐cotransport; such conditions, which were demonstrated to occur during the storage, could lead to activation of the K+Cl‐cotransport in reinfused cells. These data strongly support the evidence that K+Cl‐cotransport plays a role in K+loss and dehydration of stored erythrocytes, when incubated in autologous plasma.
Comparison of Three Different Immunoassays and PCR for the Detection of Hepatitis C Virus Infection in Pregnant Women in Taiwan
Authors: Lin Ho‐Hsiung , Kao Jia‐Horng , Leu Jer‐Herng , Young Yin‐Chin , Lee Tzu‐Yao , Chen Pei‐Jer , Chen Ding‐Shinn
Published: March 2009

To compare different hepatitis C virus (HCV) immunoassays and HCV‐RNA in pregnant women, we investigated two independent groups: 1,687 cases without screening for serum alanine aminotransferase (ALT) (group A) and 333 cases with elevated ALT (>45 IU/1) (group B), after screening 21,459 pregnant women. In group A, 11 (0.65%) and 21 (1.24%) were anti‐HCV‐positive by first‐ and second‐generation tests, respectively, while in group B 8 (2.40%) and 19 (5.71%) were positive, respectively. The results revealed by second‐generation assays based on either recombinant protein or synthetic peptides were identical, as were the anti‐HCV titers in group B. Among 40 second‐generation anti‐HCV‐positive cases, 18 (86%) of 21 in group A and 17 (89%) of the 19 in group B contained serum HCV‐RNA by RT‐PCR. Thus the prevalence of anti‐HCV in Taiwanese pregnant women is 1.24% versus 5.71% in those with elevated ALT level.
Paired Plateletpheresis with the CS 3000 Plus Blood Cell Separator Using the New 30 ml Low‐Volume Collection Chamber and the A‐35 Collection Chamber
Authors: Moog R. , Müller N.
Published: March 2009

Eight blood donors underwent paired plateletpheresis using the CS 3000 Plus blood cell separator. For each donor, two samples were run: one with the TNX‐6TM separation and the new 30‐ml low‐volume collection chamber (LVCC) and one with TNX‐6 separation and the A‐35 collection chamber. With the LVCC, 4.8±0.5 times 1011 thrombocytes could be harvested, whereas 4.4.±1.4 times 1011 platelets (PLT) were collected with the A‐35 chamber. The separation efficiencies were 51.7±6.6% for the LVCC and 43.8±10.7% for the A‐35 chamber. The leukocyte contamination of the platelet concentrates was not statistically different.
Haemolytic Anaemia to the Alpha‐lnterferon Treatment: A Proposed Mechanism
Authors: Barbolla L. , Paniagua C. , Outeiriño J. , Prieto E. , Fayos J. Sánchez
Published: March 2009

Auto‐immune haemolytic anaemia (AIHA) has been found in a case of alpha‐interferon treatment. Serum antibody and eluate were positive in the absence of the drug. Although the patient recovered after the treatment was stopped, DAGT remained positive for at least 8 months. The mechanism proposed to explain why this drug induced AIHA is similar to that proposed for alpha‐methyl‐dopa. Drugs could alter the red cell membrane and impair the immune system. Such changes have been observed with alpha‐interferon and were related with increased autoimmunity.
Prevalence and Consistency of ALT Elevation in Plasmapheresis Donors: Implications for the Assessment of Blood Product Infectivity
Authors: Picken M. , Gillon J. , Prowse C.
Published: March 2009

Alanine aminotransferase (ALT) levels were determined in 8,420 plasmapheresis donations obtained from 431 donors over a period of 18 months. Using sex‐differentiated normal ranges 2.5% of donations but 23% of donors exhibited elevated ALT levels on at least 1 occasion. Amongst the donors with elevated ALT this was only seen on 1 occasion in one third, while a quarter had elevations in consecutive donations. No donors with consecutive elevations above 100IU/1 were detected. The results are discussed in terms of the guidelines currently recommended for assessing post‐transfusion hepatitis infectivity of blood products, such as factor VIII. It is concluded that the current allowance for infection acquired from sources other than blood products under consideration may be over‐generous, leading to a potential underestimate of the true rate of infection.
Use of the MAIEA Technique to Confirm the Relationship between the Cromer Antigens and Decay‐Accelerating Factor and to Assign Provisionally Antigens to the Short‐Consensus Repeats
Authors: Petty A. C. , Daniels G. L. , Anstee D. J. , Tippett P.
Published: March 2009

The MAIEA (monoclonal‐antibody‐specific immobilisation of erythrocyte antigens) assay has recently been developed for the assignment of red cell antigens, recognised by human alloantisera, to particular membrane components of the red cell membrane [17]. This technique detects trimolecular complexes formed by the reaction of a human antibody and a mouse antibody with a particular red cell protein. A positive reaction, in an ELISA‐type detection procedure, occurs if the epitopes to the human and mouse antibodies are present on the same membrane component but at different regions. In this report, we show how the MAIEA assay can be used to confirm the relationship between Cromer system antigens and the complement‐regulatory protein, decay‐accelerating factor (DAF, CD 55). In addition, the location of the antigens along the protein is postulated by using three anti‐DAF monoclonal antibodies with specificities to different regions of DAE Tca and Esa are assigned provisionally to the first short‐consensus repeat (SCR), UMC to the second SCR, Dra to the third SCR and Cra, WESa and WESb to the fourth SCR or to the serine/threonine rich region of the DAF protein.
Cytomegalovirus Studies on Blood Donors in North‐East Scotland and a Review of UK Data
Authors: Galea G. , Urbaniak S.J.
Published: March 2009

The epidemiology of cytomegalovirus (CMV) infection as assessed by CMV antibody testing varies significantly in different parts of the world and also in different regions of a country. A study was therefore carried out to assess the prevalence of CMV antibody in our blood donors, looking in particular at different subgroups of donors as divided by age, sex and social class, to see how we could maximise the yield of CMV‐negative blood from our testing. It was shown that patterns of CMV infection in the North East of Scotland followed well‐established norms. Data from other Regional Transfusion Centres within the UK were also collected for comparison.
CD4+ Lymphocyte Depletion in HIV‐Infected Patients is Associated with gp120‐Immunoglobulin‐Complement Attachment to CD4+ Cells
Authors: Daniel Volker , Süsal Caner , Prodeus Andrej P. , Weimer Rolf , Zimmermann Rainer , Huth‐Kühne Angela , Opelz Gerhard
Published: March 2009

The mechanism of CD4+ lymphocyte depletion, which is the main immunological feature in HIV‐infected patients, is unclear. We investigated whether gp120‐immunoglobulin‐complement complexes on the surface of CD4+ cells might be involved in the elimination of CD4+ lymphocytes. The results obtained in 63 HIV‐infected patients show that gp120 is attached to a variable degree to CD4+ cells. Importantly, the percentage of CD4+gp120+ lymphocytes is inversely associated with CD4+ lymphocyte counts in the peripheral blood (p = 0.0004). CD4+gp120+ blood lymphocytes bind IgM (p = 0.0027) and IgG antibodies (p = 0.0001) and complement (p = 0.0005). These results suggest that immune complex‐mediated cell elimination is an important mechanism of CD4+ cell depletion in patients with AIDS.
Functional Cooperation among Human IgG‐Specific Murine Monoclonal Antibodies for the Detection of Weak Blood Group Antibodies in Routine Agglutination Tests
Authors: Lauren Mark , Marcil Anne , Verrette Sylvain , Lsmieux Réal
Published: March 2009

Seven monoclonal antibodies (MAbs) selected from a large panel of human IgG‐specific murine MAbs were characterized serologically and studied for their ability to cooperate in routine antihuman globulin agglutination tests. In binding inhibition experiments, three of these MAbs were shown to bind simultaneously to immobilized human IgG molecules. Cooperation among these MAbs increased significantly the capacity of the individual MAbs to agglutinate red cells sensitized with weak blood group antibodies. These results demonstrate the usefulness of selected MAb blending for the preparation of potent antihuman IgG reagents from murine monoclonal antibodies.
Evidence for the Protective Effect of Maternal FcR‐Blocking IgG Alloantibodies HLA‐DR in Rh D‐Haemolytic Disease of the Newborn
Authors: Dooren M. C. , van Kamp I. L. , Kanhai H. H. H. , Gravenhorst J. Bennebroek , von dem Borne A. E. G. Kr. , Engelfriet C. P.
Published: March 2009

In cases of Rh D alloimmunization, strong results in the antibody‐dependent cell‐mediated cytotoxicity (ADCC) assay (> 80% lysis as compared to that of the standard anti‐D serum) are indicative of severe hemolytic disease to occur in the newborn (HDN). However, discrepant cases were found in which the maternal anti‐D gave strong ADCC results and the newborns had no or only mild hemolysis. In the majority of these cases the mother had produced monocyte‐reactive IgG alloantibodies, mostly with HLA‐DR specificity. Such antibodies may be capable of blocking FcR‐mediated functions of the fetal MPS, and it has been postulated that they inhibit destruction of anti‐D‐sensitized red cells. We here describe 2 families in which such discrepancies were noticed. In 1 family, in spite of ADCC results of >80%, the Rh D‐positive second child was born without signs of hemolysis. However, the Rh D‐positive third child suffered from very severe hemolytic disease. The mother had produced monocyte‐reactive HLA‐DR antibodies in the second pregnancy which, however, did not react with the cells of the third child. In the other family, the severely Rh D‐alloimmu‐nized mother had lost her fourth child because of intrauterine death due to HDN. The Rh D‐positive fifth child was born with only mild HDN and only in this pregnancy had the mother produced such antibodies. These 2 case histories give further evidence that maternal monocyte‐reactive alloantibodies, in both these cases directed against HLA‐DR antigens, can protect the Rh D‐positive child against hemolytic disease in case of severe Rh D alloimmunization.
Plasma and Red‐Cell Glycolipid Patterns of Le(a+b+) and Le(a+b‐) Polynesians as Further Evidence of the Weak Secretor Gene Sew
Authors: Henry S. M. , Woodfield D. G. , Samuelsson B. E. , Oriol R.
Published: March 2009

Monoclonal antibodies and thin‐layer chromatography were used to study the unusual erythrocyte Lewis phenotypes found in healthy Polynesians. A single monoclonal anti‐Leb reagent 073 (clone LM129) was found which could detect Leb antigen on the Polynesian erythrocytes of samples that were unreactive with various polyclonal and monoclonal anti‐Leb reagents. Glycolipid fractions prepared from the plasma and erythrocytes of selected Polynesian samples of red‐cell Le(a‐b‐), Le(a+b‐) and Le(a+b+) phenotypes were found to have Leb glycolipids. The Leb antigen in some individuals is so weakly expressed that it is undetectable by routine erythrocyte phenotyping. Unusually large glycolipids bearing the Leb epitope were also found in some Polynesian samples, although the contribution of these novel glycolipids to phenotyping is unclear. The inability to detect Leb by routine methods and the presence of novel structures can be partially explained in terms of the presence of a weak secretor gene Sew.
HLA Antigens on Leukocyte Fragments and Plasma Proteins: Prestorage Leukoreduction by Filtration
Authors: Dzik S. , Szuflad P. , Eaves S.
Published: March 2009

HLA alloimmunization following blood transfusion results from recipient exposure to donor alloantigens. Numerous studies have documented that intact donor leukocytes are capable of provoking primary alloimmunization and that leukoreduction can decrease the incidence of primary HLA alloimmunization. HLA antigens also exist in soluble form and are present on leukocyte cell fragments. We measured the concentration of soluble HLA class I antigen in both standard and leukoreduced blood components during storage. Although the concentration of soluble class I HLA protein varied widely among different individuals, the concentration was stable during refrigerated storage of red cell concentrates and was not affected by leukocyte reduction by filtration. We also investigated whether or not HLA antigens present on leukocyte fragments were capable of stimulating either resting or in‐vitro‐primed lymphocytes in the mixed lymphocyte reaction (MLR). Leukocyte fragments prepared by repeated freeze‐thaw were found to express both class I and class II HLA antigens, but fragments were not able to stimulate in primary or secondary MLR even in cultures supplemented with recombinant interleukin‐2. These studies provide in vitro evidence to support the hypothesis that HLA antigens are not shed from intact cells to soluble forms during storage and that HLA alloantigens on cell fragments do not elicit a cellular immune response.
Transplacental IgG Subclass Concentrations in Pregnancies at Risk of Haemolytic Disease of the Newborn
Authors: Lubenko A. , Contreras Marcela , Rodeck C.H. , Nicolini U. , Savage Julia , Chana H.
Published: March 2009

The relationship of haemolytic disease of the newborn (HDN) to the transplacental passage of the four IgG subclasses was assessed at varous gestational ages by comparing the maternal and fetal IgG subclass concentrations in 34 pregnancies at risk of HDN with those in 30 pregnancies not at risk. Higher maternal and fetal IgG1 levels were attained in pregnancies at risk of HDN than in pregnancies not at risk. In contrast, a slight decrease in maternal IgG2 and IgG4 levels occurred in pregnancies at risk of HDN, as compared with a slight rise in maternal IgG2 and IgG4 levels in pregnancies not at risk of HDN. Changes in fetal IgG2 and 4 concentrations in either type of pregnancy were very similar, showing only slight increases between the 19th and 34th week of gestation. A slight decrease in maternal IgG3 occurred in both types of pregnancy. In contrast, higher and fairly steady levels of fetal IgG3 were observed in fetuses not at risk of HDN throughout gestation, when compared with those in ‘at risk’ pregnancies. However, the statistical reliability of these results is not clear since only small numbers of samples were tested and because wide variations in IgG concentrations were observed. The IgG subclass concentrations in 50 paired maternal and cord blood samples were also measured and revealed that IgG1 levels were substantially higher in cord rather than maternal blood; cord and maternal IgG2, 3 and 4 levels, on the other hand, were fairly similar.
A New Private Platelet Antigen, Groa, Localized on Glycoprotein IIIa, Involved in Neonatal Alloimmune Thrombocytopenia
Authors: Simsek Suat , Vlekke André B.J. , Kuijpers Robert W.A.M. , Goldschmeding Roel , von dem Borne Albert E. G. Kr.
Published: March 2009

The serum of a Caucasian woman who gave birth to a child with neonatal alloimmune thrombocytopenia contained antibodies directed against a platelet antigen of the newborn. There was no incompatibility for the known platelet alloantigens HPA‐1 to HPA‐7 or for the private or low‐frequency antigens Sraand Vaa, between the platelets of the parents. However, crossmatching with the serum of the mother and the platelets of the child and the father was strongly positive, suggesting a new platelet antibody specificity. To investigate the inheritance of the ‘Groa’ antigen involved, the available family members were tested in the platelet immunofluorescence test (PIFT) and the monoclonal antibody‐specific immobilization of platelet antigens (MAIPA) assay. The Groaantigen was found to be inherited in an autosomal‐codominant fashion. In the MAIPA, we localized the Groaantigen on the glycoprotein IIb/IIIa complex (αIIbβ3). The GP IIb/IIIa localization was confirmed in immunoprecipitation studies. In Western blotting experiments, we further localized the Groaantigen on the GP IIIa (β3) subunit of the GP IIb/IIIa complex. Until now we have tested approximately 400 unrelated donors. None of these appeared to be positive for the Groaantigen, suggesting a phenotype frequency in the Dutch population of less than 0.01.
Soluble HLA Class I and Beta‐2‐Microglobulin Plasma Concentrations during Interferon Treatment of Chronic Myelogenous Leukemia
Authors: Hillebrand Klaus , Moritz Thomas , Westhoff Ulrike , Niederle Norbert , Grosse‐Wilde Hans
Published: March 2009

Soluble class I molecules (sHLA‐ABC) were measured by an enzyme‐linked immunosorbent assay (ELISA) in plasma samples of 13 patients with chronic‐phase Ph1‐positive chronic myelogenous leukemia (CML). The patients were treated once daily with interferon (IFN) s.c. at a dosage of 4 times 106IU/m2IFN‐α‐2b or in combination with 50 μg IFN‐γ. Measurements were performed before 2, 4, 6, 8, 24, 48, and 72 h after the start of treatment and thereafter every 2–4 weeks. Baseline sHLA‐ABC levels were within normal limits (mean 22.1+8.8 mg/l). An initial decrease of sHLA‐ABC (mean 3.2+2.7 mg/l) was seen in all patients during the first 2–8 h of IFN treatment. Thereafter, sHLA‐ABC levels increased steadily reaching maximum values within 2–5 weeks. The overall increase was 12.7+12.4 mg/l. During the following 2–4 months of IFN treatment sHLA‐ABC decreased to near baseline levels in 12 of 13 patients. No difference was detected between IFN‐α and IFN‐α plus IFN‐γ treatment. β2‐Microglobulin values were measured in 8 patients and were found to be correlated to sHLA‐ABC concentrations (r = 0.48).
Reliable and Very Sensitive Flow‐Cytometric Method for Counting Low Leucocyte Numbers in Platelet Concentrates
Authors: Deneys V. , Mazzon A.M. , Robert A. , Duvillier H. , De Bruyère M.
Published: March 2009

Reduction of leucocytes in blood components, achieved by filtration, may reduce the risk of HLA alloimmunization, virus transmission, and febrile reactions. Nevertheless, white blood cell (WBC) concentrations obtained in this way can be too low to be accurately counted with automated and manual techniques. We describe here a rapid, reliable, and very sensitive method for counting low leucocyte numbers using flow cytometry. WBC nuclei are labelled by propidium iodide. A known amount of fluorescein‐ and phycoerythrin‐conjugated beads (Standard Brite, Coulter) is added as an indicator of the examined volume. The time required for analyzing one tube is approximately 5 min. This method was first validated by comparing WBC counts (120–2,570/μl) assessed on a haemocytometer (Bürker) and by flow cytometry. The second step was to determine the sensitivity of the method: we diluted normal platelet concentrates with phosphate‐buffered saline. The comparison of expected with observed values showed a very good correlation up to a concentration of 1 WBC/μl. This technique is an accurate method that can be applied in blood bank quality controls and in clinical studies.
Production of a Murine Monoclonal Antibody to the Low‐Incidence Red Cell Antigen Wra: Characterisation and Comparison with Human Anti‐Wra
Authors: Ring S.M. , Green C. A. , Swallow D. M. , Tippett P.
Published: March 2009

The production of a monoclonal antibody which detects the low‐incidence red cell antigen Wra is described. The antibody (BGU1‐WR) was isolated following immunization of BALB/tk mice with Wr(a+) cells. BGU1‐WR is an IgGl antibody and reacted in a manner similar to human polyclonal anti‐Wra with untreated, protease treated and chemically modified Wr(a+) cells.
Comparative Immunochemical Analysis of Wra and Wrb Red Cell Antigens
Authors: Ring S. M. , Tippett P. , Swallow D. M.
Published: March 2009

This paper describes immunoblotting and immunoprecipitation studies using monoclonal anti‐Wra and anti‐Wrb antibodies to investigate the nature of the lowincidence blood group antigen, Wra and its high‐incidence allelic antigen Wrb. No membrane components were identified by the immunoblotting experiments. Immunoprecipitation studies confirmed that the Wrb antigen involves both glycophorin A and band 3. The monoclonal anti‐Wra BGU1‐WR, failed to immunoprecipitate these or any other red cell membrane component. The significance of these findings is discussed.
Neonatal Thrombocytopenia in HLA‐DR, ‐DQ, ‐DP‐Typed Mother due to Rare Anti‐HPA‐1b (PLA2) (Zwb) Fetomaternal Immunization
Authors: Mercier P. , Chicheportiche C. , Reviron D. , Dabanian C. , Bourgue F. , Chiaroni J. , Diallo M. , Delord Y.
Published: March 2009

A new case of rare neonatal alloimmune thrombocytopenia, due to an IgG anti‐HPA‐1b in a mother HPA‐1 (a+, b‐), was diagnosed using monoclonal antibody‐specific immobilization of platelet antigens. Clinically, it was similar to the 2 previously reported observations and confirmed that, in this particular case of anti‐HPA‐1b, the treatment with random platelet pools may be as effective as selected single‐donor platelet units when maternal platelets are unusable. The HLA‐DR, ‐DQ, ‐DP genotypes of the family were obtained by PCR‐SSO. The mother's typing, compared to the HLA‐DR of the 6 similar cases reported in Europe, suggests that a combined effect of two rare HLA haplotypes might enhance this immunization.
Monoclonal IgM Cold Agglutinins with Anti‐Pr1d Specificity in a Patient with Peripheral Neuropathy
Authors: Herron B. , Willison H.J. , Veitch J. , Roelcke D. , Illis L.S. , Boulton F.E.
Published: March 2009

A patient with a demyelinating sensory motor polyneuropathy secondary to IgM paraproteinaemia is reported. The paraprotein binds to the gangliosides GD1b, GT1b, GQ1b and GD3, all of which contain disialosyl groups with the sequence NeuAcα2–8NeuAcα2–3Gal. The paraprotein also acts as a cold agglutinin recognising the sialic‐acid‐dependent Pr1d antigenic determinant of the red cell membrane glycophorins. In this and in similar cases that have been reported, the coexistence of anti‐Pr cold agglutinins and peripheral neuropathy suggest that they might be the causative agents of the disease.
Irradiation of Fresh Whole Blood for Prevention of Transfusion‐Associated Graft‐versus‐Host Disease Does Not Impair Platelet Function and Clinical Hemostasis after Open Heart Surgery
Authors: Lavee J. , Shinfeld A. , Savion N. , Thaler M. , Mohr R. , Goor D.A.
Published: March 2009

Since our previous studies suggested that the transfusion of 1 unit fresh whole blood (FWB) after cardiopulmonary bypass (CPB) using a bubble oxygenator may provide hemostatic benefit equivalent to 8–10 units of platelet concentrates, we have routinely used FWB at the termination of CPB. Two patients who received FWB and developed transfusion‐associated graft‐versus‐host disease (TA‐GVHD) prompted us to investigate the effect of irradiation of FWB on platelet and clinical hemostasis. Twenty‐four patients were randomized to receive either 1 unit FWB (12 patients), or 1 unit irradiated FWB (IrFWB, 1,500 cGy, 12 patients) after CPB. Platelet aggregation on extracellular matrix, studied by a scanning electron microscope and graded from 1 to 4 (from poor to excellent aggregation), was similar in both groups preoperatively [3.3±0.9 (FWB) and 3.5±0.5 (Ir FWB)], and at the end of CPB [1.8±1.2 (FWB) and 1.9±0.9 (IrFWB)]. Platelet aggregation was similar after transfusion of FWB (3.0±1.0) and after IrFWB (3.2±0.8), as was the increase in platelet count. Twenty‐four hours total postoperative bleeding was similar (560±420 and 523±236 ml for FWB and IrFWB, respectively). We conclude that irradiation of FWB for prevention of TA‐GVHD does not impair platelet aggregating capacity, and can be used when blood is donated by the patient's next of kin.
Flow Cytometric Analyses of the Subclasses of Red Cell IgG Antibodies
Authors: Lynen R. , Neuhaus R. , Schwarz D.W.M. , Simson G. , Riggert J. , Mayr W.R. , Köhler M.
Published: March 2009

We report on flow cytometric IgG subclass determinations of red cell antibodies using polyclonal FITC‐labeled antibodies. The limit of detection of this method was 1 ng anti‐D per 1×107 red cells. The inter‐ and intra‐assay coefficients of variance were 8.2 and 2.3%, respectively. In 8 newborns with a positive direct antiglobulin test (DAT) in the gel centrifugation test (GCT), due to ABO antibodies, IgG1 was detected in all and IgG2 additionally in 4 of these cases. In 5 severe cases of hemolytic disease of the newborn (HDN) due to anti‐D, large amounts of IgG1 were found, and in 3 of these 5, IgG3 in combination with IgG1. In 8 mild or moderate HDN cases (4 anti‐D, 2 anti‐E, 1 anti‐Fya, 1 anti‐Jka), phototherapy sufficed, and IgG1 was the only antibody. In 7 adult patients with malignant lymphoma and a positive DAT (GCT), only small amounts of IgG1 red cell autoantibodies could be demonstrated by flow cytometry. In 5 further patients with malignant lymphoma, a positive DAT, and severe hemolytic anemia, large amounts of IgG1 autoantibodies were found and IgG3 was also present in 3 of these cases. Flow‐cytometric determination of IgG subclasses may be a useful tool in immunohematology, since subclass determinations were possible in all of these cases. This method is suited for clinical routine and offers the possibility of sufficient standardization.
Anti‐N‐methyl‐d‐aspartate receptor antibody limbic encephalitis
Authors: Khadem G. M. , Heble S. , Kumar R. , White C.
Published: March 2009

We report the case of a 57‐year‐old woman who developed acute psychiatric symptoms, behavioural disturbances, insomnia and dystonia resembling a catatonic state. During the course of her illness she developed hypoventilation and required monitoring in the intensive care unit. Her serum and cerebrospinal fluid showed antibodies to the NR1/NR2 heteromers of the N‐methyl‐d‐aspartate receptor (NMDAR). Anti‐NMDAR encephalitis is a severe form of autoimmune encephalitis, which has only recently been described in the published work. Most patients improve with immunosuppressive treatment. Raising awareness of this rare but increasingly reported condition is important, as it is responsive to treatment and potentially reversible.