bluuebunny/crossref_metadata_2025 huggingface dataset:
Virus reduction in the preparation of intravenous immune globulin: in vitro experiments
Authors: Chandra Sudhish , Cavanaugh James E. , Lin Chaomei M. , Pierre‐Jerome Claudette , Yerram Nagender , Weeks Richard L. , Flanigan Everett , Feldman Fred
Published: March 2003

BACKGROUND: While immune globulins for intravenous administration (IGIV) have an excellent record with respect to virus safety, concern regarding these preparations has been raised by reports of transmission of hepatitis C virus (HCV) to patients treated with IGIV and the presence of genetic material for HCV in IGIV preparations.

STUDY DESIGN AND METHODS: This in vitro study evaluated the effectiveness of several manufacturing steps, including ethanol precipitation and pasteurization, in reducing HIV and model viruses including encephalomyocarditis (EMC) virus, pseudorabies virus (PRV), bovine viral diarrhea virus (BVDV), Sindbis virus, vaccinia virus, and vesicular stomatitis virus (VSV), as well as HCV RNA, in IGIV.

RESULTS: Ethanol precipitation carried out after pasteurization resulted in virus reductions (log10) of >3.97 for HIV, 1.95 for EMC virus, >5.39 for PRV, and 3.52 for BVDV. Pasteurization inactivated EMC virus by 4.52 log10 and resulted in a log10reduction of >6.54 for HIV, >5.39 for PRV, >6.64 for BVDV, >7.78 for Sindbis virus, >5.84 for vaccinia virus, and >6.99 for VSV. All viruses except EMC virus were reduced below the limit of detection within 6 hours of the beginning of pasteurization. Cohn processing of Fraction II + III paste and the 4.5‐percent alcohol precipitation step prior to pasteurization provided additional virus removal. Studies using the polymerase chain reaction technique found that HCV RNA was detectable in the starting fraction of Cohn Fraction II paste, but not in the final IGIV preparation.

CONCLUSION: These findings strongly support the viral safety of IGIV prepared by this method and show a significant added measure of virus safety associated with pasteurization of this preparation.
Distinguishing immunosilent AIDS from the acute retroviral syndrome in a frequent blood donor
Authors: Kopko Patricia M. , Calhoun Loni , Petz Lawrence D.
Published: March 2003

BACKGROUND: There are seven reports of “immunosilent AIDS” in which there was a lack of development of anti‐HIV for more than 6 months. Thus, when a frequent blood donor presented with clinical findings highly suggestive of overt AIDS, there was concern that he may have had a prolonged immunosilent infection.

CASE REPORT: A 24‐year‐old man who had donated blood six times in the previous year was diagnosed as having AIDS; he presented with fever, nausea, vomiting, diarrhea, weight loss, and oral candidiasis. The anti‐HIV enzyme immunoassay was positive, the Western blot was indeterminate (gp160 only), the CD4+ count was 174 per mL, the HIV polymerase chain reaction was positive (2.8 × 106 copies/mL), and the HIV p24 antigen assay was positive. Twelve components from previous donations had been transfused, and 2 units of fresh‐frozen plasma were still in inventory. Repeat donor testing 57 days after donation indicated seroconversion with a positive anti‐HIV enzyme immunoassay, a positive Western blot, a negative HIV p24 antigen assay, and a positive test for HIV by polymerase chain reaction (89,000 copies/mL). Both units of fresh‐frozen plasma tested negative for HIV by polymerase chain reaction. Four transfusion recipients had died, and the remaining eight are anti‐HIV negative with >6 months' follow‐up.

CONCLUSION: The donor had an unusually severe acute retroviral syndrome and presented with findings that were difficult to distinguish from overt AIDS.
The value of oxygen‐carrying solutions in the operative setting, as determined by mathematical modeling
Authors: Brecher Mark E. , Goodnough Lawrence T. , Monk Terri
Published: March 2003

BACKGROUND: The use of oxygen carriers (red cell [RBC] substitutes) in acute trauma and in surgery, with or without the use of acute normovolemic hemodilution (ANH), is being investigated. Mathematical modeling was used to assess the impact of RBC substitutes, with or without ANH, in the elective surgical setting.

STUDY DESIGN AND METHODS: Mathematical equations and computer models were developed on the basis of previously described mathematical principles, for better understanding of the potential efficacy of RBC substitutes for blood needs with or without ANH. Savings were calculated for a patient with a blood volume of 5000 mL and an initial hematocrit (Hct) of 45 or 30 percent.

RESULTS: Substantial increases in the tolerable blood losses (or reduced allogeneic RBC needs) were most evident when the use of an RBC substitute to achieve severe ANH to a Hct that the patient might not otherwise have been able to tolerate was combined with the use of RBC substitutes as replacement for the surgical blood subsequently lost. However, the benefit was greatly dependent on the patient's initial Hct. For example, for a patient with a blood volume of 5000 mL and an initial Hct of 45 percent, a blood loss of approximately 2500 mL resulted in a final Hct of 28 percent without the use of an RBC substitute or ANH. In contrast, with the combined use of staged ANH with an RBC substitute and the RBC substitute for lost surgical blood, a blood loss of up to 14.5 L could be tolerated. However, in an anemic patient (blood volume 5000 mL, initial Hct 30%), a Hct of 28 percent cannot be sustained without the use of allogeneic RBCs for any of the described strategies, even when blood losses were as low as 1 L.

CONCLUSION: The use of RBC substitutes has the potential to result in a substantial reduction in allogeneic RBC exposure. This benefit is essentially limited to the nonanemic patient when the use of an RBC substitute is combined with severe ANH and there is concomitant large perioperative blood loss. Anemic patients can be expected to have only limited benefit, because of an inability to sequester an adequate volume of autologous RBCs via ANH.
Immune hemolytic anemia induced by 6‐mercaptopurine
Authors: Pujol M. , Fernández F. , Sancho J.M. , Ribera J.M. , Millá F. , Feliu E.
Published: March 2003

BACKGROUND: Myelosuppression is the main hematotoxic effect of 6‐mercaptopurine (6‐MP), which is an antimetabolite chemotherapy drug. Immune hemolytic anemia associated with this drug has not been previously reported.

CASE REPORT: A 67‐year‐old man with chronic myelomonocytic leukemia presented with anemia 2 weeks after 6‐MP therapy had been initiated. Additional tests provided laboratory evidence of hemolysis. When treatment was stopped, the patient's condition and laboratory results showed a progressive improvement.

RESULTS: The direct antiglobulin test was positive for IgG. The eluate and the serum were not reactive with panel red cells but reacted with 6‐MP‐treated red cells, while the normal serum pool was unreactive. The direct antiglobulin test was no longer positive by 20 days after the cessation of 6‐MP therapy.

CONCLUSION: This drug, 6‐MP, should be added to the list of drugs that have been reported to cause immune hemolytic anemia by means of the so‐called hapten mechanism.
Neutrophil antigen 5b is carried by a protein, migrating from 70 to 95 kDa, and may be involved in neonatal alloimmune neutropenia
Authors: De Haas M. , Muniz‐Diaz E. , Alonso L.G. , Van Der Kolk K. , Kos M. , Buddelmeijer L. , Porcelijn L. , Von Dem Borne A.E.G.Kr.
Published: March 2003

BACKGROUND: Neutrophil antigen 5b has been described as involved in transfusion reactions and not in neonatal alloimmune neutropenia.

CASE REPORT: Anti‐5b was found in the serum of a mother of a persistently neutropenic newborn, who had several bacterial infections. The neutropenia responded to treatment with recombinant human granulocyte–colony‐stimulating factor. Immunoprecipitation experiments performed with this and three other 5b antisera identified a protein, migrating from 70 to 95 kDa, as carrier of 5b. The observed pattern of migration may point to heavy glycosylation of this protein.

RESULTS: Six 5b‐negative donors were identified among 54 screened white donors, for a 5b gene frequency of 0.66.

CONCLUSION: Alloimmunization to 5b in pregnancy is rare. In the patients with neonatal neutropenia analyzed in the last decade, this was the first case discovered.
Regeneration of erythropoiesis after related‐ and unrelated‐donor BMT or peripheral blood HPC transplantation: a major ABO mismatch means problems
Authors: Worel N. , Greinix H.T. , Schneider B. , Kurz M. , Rabitsch W. , Knöbl P. , Reiter E. , Derfler K. , Fischer G. , Hinterberger W. , Höcker P. , Kalhs P.
Published: March 2003

BACKGROUND: Blood group incompatibility in allogeneic BMT is common but does not appear to affect the outcome in terms of incidence of graft rejection or delayed engraftment. However, major ABO incompatibility may be associated with prolonged erythroid aplasia.

STUDY DESIGN AND METHODS: In a retrospective analysis of 286 allogeneic transplant recipients, the prevalence of prolonged erythroid aplasia, including pure RBC aplasia, was determined.

RESULTS: Patients receiving major ABO‐incompatible grafts showed a significant delay in reticulocyte engraftment (median, 32 days; range, 12‐347) from that in patients receiving ABO‐identical (20; 10‐152) or minor ABO‐incompatible (21; 12‐47) grafts. Pure RBC aplasia occurred in 7 (3%) of 240 evaluable recipients and was observed only in the major ABO‐incompatible group (7/43, 16%). Treatment of pure RBC aplasia consisted of either plasma exchange, which resulted in a response within 16 to 68 days, or immunoadsorption, in which the response occurred between Days 119 and 204 after initiation of treatment.

CONCLUSION: Major ABO incompatibility may lead to delayed reticulocyte engraftment, resulting in prolonged transfusion dependency and increased risks of transmission of infection and iron overload. Therefore, therapeutic strategies should be taken into consideration to allow erythroid reconstitution in these patients.
Alloimmunization in preterm infants after repeated transfusions of WBC‐reduced RBCs from the same donor
Authors: Strauss Ronald G. , Johnson Karen , Cress Gretchen , Cordle Delores G.
Published: March 2003

BACKGROUND: Preterm infants are among the most heavily transfused of patient groups, yet multiply transfused infants only rarely produce alloantibodies against RBC or WBC antigens. It is not known whether rates of alloimmunization might be increased by repeated exposure to RBCs and WBCs from the same donor, as in limited‐donor‐exposure programs, or whether infants might benefit from WBC‐reduced RBC components as a means of diminishing the risk of possible alloimmunization.

STUDY DESIGN AND METHODS: Preterm infants (birth weight 0.6‐1.3 kg) received prestorage WBC‐reduced RBCs from dedicated donors, collected in AS‐3 as a means of limiting donor exposures. Blood samples were collected serially from infants shortly after birth until either discharge or age 6 months and were studied for RBC and WBC antibodies—the latter with reactivity against either HLA class I or neutrophil‐specific antigens.

RESULTS: Thirty preterm infants received 139 transfusions (mean, 4.6; median, 4 transfusions per infant), with 81 percent of transfusions obtained from one donor per infant. Eighty‐four blood samples (mean, 2.7/infant) were studied, and no infant produced RBC antibodies. Twenty‐seven percent of infants exhibited WBC antibodies, but only 13 percent actually produced WBC antibodies (passive maternal antibody excluded). Of the WBC antibodies produced by infants, three were against HLA class I and one was against neutrophil‐specific antigens; none were linked to adverse effects.

CONCLUSIONS: Because infants only rarely produce RBC antibodies, no changes in blood banking practices are necessary for limited‐donor‐exposure programs. Although the production of WBC antibodies by infants occurs, it seems to be uncommon; thus, the possible benefits, if any, of WBC reduction are uncertain, and further study is required before changes in practice can be justified.
A new h allele detected in Europe has a missense mutationin α(1,2)‐fucosyltransferase motif II
Authors: Wagner Thomas , Vadon Maria , Staudacher Erika , Schmarda Andreas , Gassner Christoph , Helmberg Wolfgang , Lanzer Gerhard , Flegel Willy A. , Wagner Franz F.
Published: March 2003

BACKGROUND: The FUT1 gene encodes an α(1,2)‐fucosyltransferase (H transferase), which determines the blood group H. Nonfunctional alleles of this gene, called h alleles and carrying loss‐of‐function mutations, are observed in the exceedingly rare Bombay phenotype. Twenty‐three distinct h alleles have been characterized at the molecular level in various populations. The FUT2 (SE) gene is highly homologous to FUT1 (H).

STUDY DESIGN AND METHODS: The FUT1 gene of an Austrian proband with the Bombay phenotype was characterized by nucleotide sequencing of the full‐length coding sequence. A PCR method using sequence‐specific primers for FUT2 genotyping in whites was developed. The plasma α(1,2)‐fucosyltransferase activity was determined. The distribution of the mutations underlying 24 h alleles and 7 se alleles was analyzed.

RESULTS: The proband carried a new h allele. Two nucleotide changes, G785A and C786A, in codon 262 of the FUT1 gene resulted in the replacement of serine by lysine. No α(1,2)‐fucosyltransferase activity was detected in the proband's plasma. The proband was homozygous for the seG428A allele. Six of 17 missense mutations in nonfunctional h and se alleles occurred in highly conserved fucosyltransferase motifs. No loss‐of‐function mutation was observed in the aminoterminal section encompassing the transmembraneous helix.

CONCLUSION: The missense mutation S262K in the FUT1 gene caused the loss of H transferase activity. The analysis of the distribution of mutations in nonfunctional FUT1 and FUT2 genes can point to functionally important domains in the H transferase.
Long‐term survival in transfusion recipients in Sweden, 1993
Authors: Tynell Elsa , Norda Rut , Shanwell Agneta , Björkman Anders
Published: March 2003

BACKGROUND: Survival and characteristics of transfusion recipients have not been studied enough, although they represent key measures in cost‐effectiveness analyses of various donor screening procedures.

STUDY DESIGN AND METHODS: Hospital and blood bank records were collected on all patients in Örebro County, Sweden, from March through May 1993 (1111 transfusion episodes) and a random sample from Stockholm County during April 1993 (793 transfusion episodes). All patients were then matched with the national register of deaths in Sweden during a follow‐up period of 40 months.

RESULTS: The median patient age was 71 years and the median transfusion total was 2 units. Only 35 percent of the patients were under the age of 65, 9 percent under 40, and 1.6 percent under 1 year. About half (56%) were women. Among the Örebro patients, 47 percent were surgical and 29 percent internal medicine patients. Of 1720 patients whose survival could be investigated, 66 percent were alive after 1 year and 51 percent after 40 months. The survival rates were rather similar in patients receiving RBCs and plasma but lower in those receiving platelets.

CONCLUSION: The survival of patients transfused in Sweden in 1993 differered significantly from estimations based on studies from the 1980s. This difference has major implications for the estimations of cost‐effectiveness of blood donor screening for infectious agents.
Human platelets express gangliosides with LKEactivity and ABH blood group activity
Authors: Cooling Laura L.W. , Zhang DeSheng , Koerner Theodore A.W.
Published: March 2003

BACKGROUND: Platelets express several neutral glycosphingolipids with ABH and P blood group activity that may play a role in infectious, autoimmune, and alloimmune thrombocytopenia. In RBCs, sialylated glycosphingolipids or gangliosides with blood group activity have also been reported. To determine whether similar antigens are expressed by platelets, the total platelet ganglioside fraction was isolated and screened for blood‐group‐active glycosphingolipids.

STUDY DESIGN AND METHODS: Platelet gangliosides were isolated by organic extraction, base hydrolysis, anion exchange, silicic acid, and high‐performance liquid chromatography. Gangliosides were identified and characterized by high‐performance thin‐layer chromatography‐immunostaining with blood group‐specific MoAbs and glycosidase digestion.

RESULTS: Group A, but not group O, platelets express five gangliosides with group A activity. Of five A MoAbs and lectins examined, only MoAbs Birma‐1 and MHO4 recognized all five sialyl A bands. The sialyl A bands were sensitive to endoglycoceramidase and neuraminidase. One sialyl A band may represent a branched ganglioside with sialyl‐I and group A activity. Platelets also express an LKE‐active ganglioside consistent with sialyl‐galactosylgloboside.

CONCLUSION: In addition to sialyl‐iI and sialyl‐Lex gangliosides, group A platelets express gangliosides with LKE activity and group A activity. Like RBCs, group A‐active gangliosides may act as alloantigens and autoantigens to naturally occurring isohemagglutinins.
Analysis of the kinetics of TPOuptake during platelet transfusion
Authors: Folman Claudia C. , De Jong Shreyas M. , De Haas Masja , Von dem Borne Albert E.G.Kr.
Published: March 2003

BACKGROUND: It has been shown in several studies that platelets play a role in the removal of TPO from the circulation. For instance, in vitro studies have shown that platelets can bind and internalize TPO, and transfusion studies have shown that the concentration of circulating TPO decreased after platelet transfusion. In the current study, the in vivo kinetics of plasma TPO levels and TPO uptake by transfused platelets is analyzed in more detail.

STUDY DESIGN AND METHODS: Serial blood samples from patients who received a platelet transfusion were analyzed with respect to platelet count, plasma TPO concentration, and TPO content per platelet. In addition, the capacity of transfused platelets to bind TPO in vitro was assessed.

RESULTS: Platelet counts increased immediately after transfusion, but subsequently started to decrease. Conversely, TPO levels decreased significantly but then returned to baseline level by 44 hours after transfusion. Platelet count and plasma TPO concentration were inversely correlated (rp = −0.9; p<0.05). The decrease in TPO concentration upon transfusion was accompanied by a significant increase in the platelet‐associated TPO concentration. After transfusion, platelets isolated from the patient still displayed functional TPO receptors, as indicated by their intact capacity to bind TPO in vitro.

CONCLUSION: The decrease in plasma TPO followed by the increase in platelet TPO provides evidence that platelets are responsible for the clearance of TPO in circulation. In vivo, platelets can bind and may degrade TPO upon platelet transfusion.
In vitro and in vivo measurementsof gamma‐radiated, frozen, glycerolized RBCs
Authors: Valeri C. Robert , Pivacek Linda E. , Cassidy George P. , Ragno Gina
Published: March 2003

BACKGROUND : Transfusion‐associated GVHD results from the presence of viable lymphocytes in transfused allogeneic blood components. Viable immunocompetent lymphocytes have been detected in RBCs that were frozen with glycerol and washed before transfusion.

STUDY DESIGN AND METHODS : The study reported here assessed the effect of irradiation on human RBCs frozen with 40‐percent (wt/vol) glycerol and stored at –80°C. In vitro and in vivo testing was done on human RBCs that were frozen with 40‐percent (wt/vol) glycerol at –80°C, with some units exposed to 2500 cGy of gamma radiation and others not irradiated, and that, after thawing and washing, were stored in a sodium chloride‐glucose solution at 4°C for 3 days before autologous transfusion.

RESULTS : The glycerol‐frozen RBCs treated with 2500 cGy before deglycerolization had a mean freeze‐thaw‐wash recovery of 87 percent and a mean 24‐hour posttransfusion survival of 86 percent after storage for 3 days at 4°C in a 0.9‐percent NaCl and 0.2‐percent glucose solution. For the nonirradiated units, the mean freeze‐thaw‐wash recovery was 85 percent and the mean 24‐hour posttransfusion survival was 83 percent.

CONCLUSION : These data show similar, acceptable results for RBCs frozen with 40‐percent (wt/vol) glycerol at –80°C and treated in the frozen state with 2500 cGy of gamma radiation and for RBCs that were not irradiated, all of which were washed and then stored in a sodium chloride‐glucose solution for 3 days before autologous transfusion.
The effect of external pressure, catheter gauge, andstorage time on hemolysis in RBC transfusion
Authors: Frelich Regina , Ellis Martin H.
Published: March 2003

BACKGROUND: RBC transfusions are often administered using externally applied pneumatic pressure. The effect of this practice on RBC hemolysis was studied during simulated transfusion performed in the laboratory.

MATERIALS AND METHODS: RBC transfusions were performed sequentially via 16‐, 18‐, 20‐, and 22‐gauge cannulas with the application of a pneumatic pressure device at pressures of 150 and 300 mmHg. Hb concentration, Hct, RBC count, free Hb, potassium, and LDH concentrations were measured in 5 mL of transfusate.

RESULTS: Forty‐seven RBC units, 20 fresh units (mean age, 10.8 days) and 27 old units (mean age, 28.9 days) were studied under all conditions. Multiple regression analysis revealed that the age of the unit and the external pressure applied were significant determinants of the degree of hemolysis.

CONCLUSIONS: The application of an external pressure device results in minimal destruction of transfused RBCs, even under the most stringent conditions examined. Thus, external pressure application to expedite an RBC transfusion is likely to be a safe procedure for the majority of patients. Unusual clinical situations, such as massive transfusion in pediatric patients, should be specifically examined to confirm the safety of this procedure.
The effect of the interruption of agitation on plateletquality during storage for transfusion
Authors: Hunter Suzanne , Nixon Janice , Murphy Scott
Published: March 2003

BACKGROUND: A considerable amount of data and the CFR suggest that platelet concentrates (PCs) should be stored with continuous, gentle agitation before transfusion. However, there are only limited data concerning the mechanisms of platelet damage that may occur when agitation is interrupted, and there are no CFR guidelines concerning shipment between periods of storage.

STUDY DESIGN AND METHODS: PCs were prepared by the platelet‐rich plasma method and stored for 5 days at 20 to 24°C; agitation was interrupted for 1 to 3 days either by simply stopping the agitator or by placing the PCs in a stationary shipping container. Measurements of platelet metabolism and quality were made during storage and on Day 5.

RESULTS: With interruption on the agitator, the production of lactic acid was increased during the interruption in proportion to the number of platelets in the PC and the duration of the interruption. The pO2 was increased during agitation interruption, which suggested a decline in oxygen utilization. With the use of the hypotonic shock response and the extent of shape change as reflections of platelet quality, there was no evidence of platelet damage unless the pH fell to or below 6.5. No PC reached this level after an interruption of agitation for only 1 day, irrespective of which day was chosen for interruption. PCs whose agitation was interrupted for 2 and 3 days were at risk of having a pH less than 6.5 if their contents were greater than 1.25 × 1011 and 0.75 × 1011 platelets, respectively. Interruption of agitation for 1 day in the shipping container produced results essentially identical to those produced by interruption on the agitator.

CONCLUSION: Interruption of agitation of PCs for 1 day, either on the agitator or in the shipping container, produces no platelet damage measurable by these in vitro techniques. However, an interruption of agitation for 2 days can result in significant damage in some components. Further studies will be required to learn more about the mechanisms that lead to the metabolic changes described and to determine if the same generalizations apply to apheresis PCs and PCs prepared from pooled buffy coats.
Ceftizoxime‐induced hemolysis secondary to combined drug adsorption and immune‐complex mechanisms
Authors: Calhoun Beverly W. , Junsanto Tipsuda , Donoghue Maria De Tolve , Naureckas Edward , Baron Joseph M. , Baron Beverly W.
Published: March 2003

BACKGROUND: Immune hemolytic anemia has been associated with the administration of various antibiotics, including cephalosporins. Presented here is a patient who developed severe acute hemolysis while receiving ceftizoxime (Ceftizox, Fujisawa USA), a third‐generation cephalosporin. This is the fourth reported case of hemolysis in association with ceftizoxime. In the previous cases, ceftizoxime was shown to induce hemolysis by the immune‐complex mechanism. However, in one of those reports, the concentration of drug used to treat the target RBCs in vitro may not have been optimal.

CASE REPORT: The patient's antemortem blood samples were analyzed retrospectively for drug‐dependent antibodies by the drug‐adsorption and immune‐complex methods. Antibody class and titer were evaluated.

RESULTS: The patient's sample agglutinated RBCs coated with ceftizoxime as well as uncoated RBCs in the presence of ceftizoxime. The antibodies to ceftizoxime were IgM and IgG.

CONCLUSION: This is the first report on both the immune‐complex and drug‐adsorption mechanisms of ceftizoxime‐induced hemolysis. The differential diagnosis of a falling Hct in a patient receiving antibiotics should include drug‐related hemolysis; once this diagnosis is considered, management includes the appropriate serologic workup, immediate cessation of the implicated drugs, and possible transfusion support.
Knowledge of HIV/AIDS transmission and screening in United States blood donors
Authors: Sharma Usha K. , Schreiber George B. , Glynn Simone A. , Nass Catharie C. , Higgins Martha J. , Tu Yongling , Bethel Jim , Williams Alan E. ,   for the Retrovirus Epidemiology Donor Study
Published: March 2003

BACKGROUND : Increased knowledge of HIV transmission and behavioral and test screening may encourage high‐risk blood donors to self‐defer.

STUDY DESIGN AND METHODS: Knowledge of HIV transmission and screening and the association with demographics, screening test reactivity, and unreported deferrable risks (UDRs) was assessed by a 1998 anonymous mail survey sent to 92,581 blood donors, of whom 57 percent responded. Groups were compared by using weighted chi‐square tests and logistic regression analysis.

RESULTS: Four percent of the donors thought that it was very likely or somewhat likely for a person to contract HIV from donating blood, and 20 percent perceived a similar risk from blood transfusion. Only 60 percent of the donors knew that the available screening tests may not detect a recent infection. Thirty‐seven percent either did not know or felt it was acceptable to donate blood to obtain HIV testing. Those most likely to answer knowledge questions incorrectly were more likely to have a higher prevalence of test reactivity or UDRs and to be <25 years old, members of a minority, less educated, non‐US‐born, or first‐time donors.

CONCLUSION : Innovative ways to increase donor understanding of the implications of risk behaviors and the limitations of current screening procedures should be developed. They might prevent high‐risk individuals from donating blood.
Assessment of rapid remobilization intervals with G–CSF and SCF in murine and rhesus macaque models
Authors: Shi Patricia A. , Pomper Gregory J. , Metzger Mark E. , Donahue Robert E. , Leitman Susan F. , Dunbar Cynthia E.
Published: March 2003

BACKGROUND: Defining the optimum regimen and time for repeat peripheral blood progenitor cell mobilization would have important clinical applications.

STUDY DESIGN AND METHODS: Remobilization with SCF and G–CSF at 2 weeks after an initial mobilization in mice and at 2 or 4 weeks after an initial mobilization in nonhuman primates was examined. In mice, competitive repopulation assays were used to measure long‐term progenitor cell‐repopulating activity. In monkeys, mobilization of hematopoietic progenitor CFUs was used as a surrogate marker for progenitor cell‐repopulating ability.

RESULTS: Efficacy of progenitor cell remobilization differed in the two animal species. In mice, peripheral blood progenitor cell‐repopulating ability with repeat mobilization at 2 weeks was 70 percent of that with the initial mobilization. In monkeys, there was no significant difference in peripheral blood progenitor cell mobilization between the initial and the repeat mobilizations at 2 weeks. In mobilizations separated by 4 weeks, however, peripheral blood progenitor cell mobilization was higher than that with initial mobilizations.

CONCLUSION: In animal models, mobilization of peripheral blood progenitor cells with remobilization after a 2‐week interval is similar to or moderately decreased from that with the initial mobilization. Progenitor cell collection at this time point may be useful in certain clinical circumstances. A 4‐week interval between remobilizations may be preferable. Clinical trials in humans would be useful to clarify these issues.
Increased RBC autoantibody production in pregnancy
Authors: Hoppe Berthold , Stibbe Werner , Bielefeld Annette , Pruss Axel , Salama Abdulgabar
Published: March 2003

BACKGROUND: Autoimmunization against RBCs is generally believed to occur very rarely during pregnancy and to represent a high risk for those affected. The occurrence of benign RBC autoantibodies in pregnancy is reported.

STUDY DESIGN AND METHODS: The incidence of autoantibodies to RBCs in unselected pregnant and nonpregnant women were compared by the new standard gel hemagglutination method.

RESULTS: Data from a total of 22,782 pregnant and 12,903 nonpregnant women were analyzed. The antibody screen test was positive in 547 (2.40%) of the pregnant women. The positive test results were related to passive anti‐D in 341 cases (1.50%), to clinically relevant alloantibodies in 43 cases (0.19%), to clinically irrelevant antibodies in 138 cases (0.61%), to autoantibodies in 23 cases (0.1%), and to autoantibodies plus alloantibodies in 2 cases (0.01%). In the cases of alloantibodies and/or autoantibodies, the DAT was invariably positive with anti‐C3d and less commonly so with anti‐IgG. The autoantibodies were first detected during pregnancy, they predominantly reacted with enzyme‐treated RBCs, and none of the affected women or their infants appear to have developed clinically significant hemolysis. In the control group consisting of nonpregnant female blood donors, there were 3 cases (0.02%) of clinically insignificant IgG autoantibodies.

CONCLUSION: There is an increase in autoimmunization against RBCs during pregnancy. The resultant autoantibodies do not appear to cause significant RBC destruction. Further studies are required for precise characterization of this phenomenon.
Quantitation of residual WBCs in filtered blood components by high‐throughput, real‐time kinetic PCR
Authors: Lee Tzong‐Hae , Wen Li , Chrebtow Vera , Higuchi Russell , Watson Robert M. , Sninsky John J. , Busch Michael P.
Published: March 2003

BACKGROUND: The effort to eliminate transfusion complications associated with WBCs has led to the widespread use of filters able to reduce WBC concentrations to ≤0.1 WBC per μL blood. This has necessitated sensitive QC methods to quantitate residual WBCs in filtered units. One fast, effective method is DNA amplification using real‐time kinetic PCR (kPCR).

STUDY DESIGN AND METHODS: Two methods of preparation of standards were compared and used for the optimization of quantitative kPCR. The first involved spiking genomic DNA cell lysate into a diluent, followed by a series of 1 in 10 dilutions. The second involved spiking serial 1 in 10 dilutions of WBCs into twice‐filtered fresh whole blood. Two hundred fifty filtered frozen whole‐blood samples were amplified in duplicate to show the kPCR assay's reproducibility. Another 359 filtered frozen whole blood samples were used to compare data from kPCR with data from a standard PCR protocol using 32P‐labeled probe and autoradiography. All specimens were amplified for conserved HLA DQα sequences.

RESULTS: Standards prepared by both methods gave reproducible and equivalent results. Quantitation of standards representing a dynamic range of 8 × 100 to 8 × 105 WBCs per mL, yielded standard deviations ranging from 0.59 cycle to 1.04 cycles (a one‐cycle increase is equivalent to a twofold increase in WBC concentration). The scatter graph of the 250 samples tested in duplicate by kPCR generated a slope of 1.0122 and an R2 value of 0.9265. The comparison of kPCR and 32P‐probe hybridization results on 359 clinical samples gave a scatter‐graph slope of 0.9428 and an R2 value of 0.8718, indicating excellent agreement of the methods over a 4‐log dynamic range.

CONCLUSION: kPCR is a high‐throughput, sensitive assay that could prove useful in routine quality assurance of the WBC reduction process.
Signs and symptoms associated with the transfusion of WBC‐reduced RBCs and non‐WBC‐reduced RBCs in patients with anemia and HIV infection: results from the Viral Activation Transfusion Study
Authors: Lane Thomas A. , Gernsheimer Terry , Mohandas Kala , Assmann Susan F.
Published: March 2003

BACKGROUND: RBC transfusion is associated with fever and other reactions in some patients. The Viral Activation Transfusion Study randomly assigned patients to receive either unmodified or WBC‐reduced RBCs and thus offered an opportunity to assess the effect of WBC‐reduced RBCs on the incidence of transfusion reactions prospectively.

STUDY DESIGN AND METHODS: This prospective, randomized, double‐blind, multicenter study compared prestorage WBC‐reduced RBCs to unmodified RBCs in HIV‐infected, CMV‐seropositive, and transfusion‐naive persons who required transfusions for anemia. Primary endpoints were survival and change in the plasma HIV RNA level at 7 days after transfusion. The incidence of transfusion reactions was prospectively evaluated.

RESULTS: The two groups had similar baseline characteristics and study endpoints; 3864 RBC units (median storage age, 9 days) were administered to 531 patients during 1745 transfusions. The most frequent signs reported were elevated temperature and hypotension. Subjects who reported fever within the week prior to transfusion were more likely to have an elevation in temperature associated with transfusion. The administration of RBCs that were less than 10 days old was associated with a marginal increase in the incidence of transfusion‐associated temperature elevation among recipients of unmodified RBCs, but not among recipients of WBC‐reduced RBCs. Caregivers reported fewer instances of both elevated temperature and hypotension than were identified by review of transfusion records.

CONCLUSIONS: The incidence of elevated temperature and hypotension associated with transfusion in this population was unexpectedly high. Use of WBC‐reduced RBCs had no effect on the overall rates of elevated temperature or hypotension associated with transfusion of RBCs. The occurrence of a pre‐existing fever was associated with a higher frequency of transfusion‐associated elevated temperature.
Potential value of Plasmodium falciparum‐associated antigen and antibody detection for screening of blood donors to prevent transfusion‐transmitted malaria
Authors: Silvie Olivier , Thellier Marc , Rosenheim Michel , Datry Annick , Lavigne Patricia , Danis Martin , Mazier Dominique
Published: March 2003

BACKGROUND: Malaria antibody detection is a valuable tool in the prevention of transfusion‐transmitted malaria in countries with a high proportion of donors with travel exposure to malaria. The immunofluorescent antibody test (IFAT) is still the reference method, but it is not suitable for screening of blood donors. ELISA would be an interesting alternative to the IFAT, but it lacks sensitivity.

STUDY DESIGN AND METHODS: To evaluate the potential value of a combined screening strategy based on malaria antigen and antibody detection, plasma samples from 203 patients infected with Plasmodium falciparum were tested with an ELISA for the detection of malaria antibodies (Malaria IgG CELISA, Cellabs) and a P. falciparum histidine‐rich protein‐2 kit (Malaria P.f., ICT Diagnostics) for the detection of malaria antigens.

RESULTS: Among patients with positive IFAT results, CELISA had a sensitivity of 71 percent, whereas the combined screening tests (CELISA and Malaria P.f.) had a sensitivity of 88 percent (p < 0.001). Sequential samples from 50 patients were tested. The combined screening tests shortened the detection of seroconversion from 11.4 ± 1.6 to 5.3 ± 1.1 days (p < 0.001).

CONCLUSION: Combined malaria antigen and antibody detection, with methods compatible with mass screening, may constitute an attractive alternative to the IFAT for blood donor screening.
Reduced frequency of blood donors with false‐positive HIV‐1 and ‐2 antibody EIA reactivity after elution of low‐affinity nonspecific natural antibodies
Authors: Bouillon Marlene , Aubin Éric , Roberge Chantal , Bazin Renée , Lemieux Réal
Published: March 2003

BACKGROUND : The detection by EIA of antibodies (Abs) specific to HIV antigens in the serum of blood donors is important for transfusion safety. A small but significant number of donor sera (0.1‐0.3%) yield false‐positive results in EIA, and these donors must be permanently deferred from the blood donor list, causing operational and public relations problems.

STUDY DESIGN AND METHODS : False‐positive EIA reactions could be caused by the binding of low‐affinity natural polyreactive Abs, which could be eluted by treatment with a chaotropic agent such as thiocyanate (SCN). The effect of the SCN elution on EIA specificity was studied using tetanus toxoid and HIV Abs.

RESULTS : SCN elution preferentially reduced the binding of nonspecific Abs. Testing of 235 seronegative samples in HIV‐1 and ‐2 EIA showed that SCN elution reduced the mean and dispersion of OD values obtained. In addition, SCN elution abolished the false‐ positive reactivity in HIV‐1 and ‐2 EIA of 69 percent (299/435) of the repeatedly reactive specimens tested without impact on the reactivity of HIV‐seropositive specimens.

CONCLUSION : The SCN elution step increased the specificity of HIV‐1 and ‐2 EIA and could facilitate the re‐entry of previously deferred donors.
PCR with sequence‐specific primer‐based simultaneous genotyping of human platelet antigen‐1 to ‐13w
Authors: Lyou Jau‐Yi , Chen Ying‐Ju , Hu Hui‐Yu , Lin Jeong‐Shi , Tzeng Cheng‐Hwai
Published: March 2003

BACKGROUND : Accurate human platelet antigen (HPA) typing is important for patients with diagnosis of alloimmune thrombocytopenic syndromes and provision of HPA‐matched blood components for these patients.

STUDY DESIGN AND METHODS : Thirteen sequence‐specific primers (SSPs) designed on the basis of known published polymorphisms for HPA‐1 to HPA‐13w, respectively, were employed for simultaneous HPA genotyping. All PCR amplifications were carried out with identical cycling conditions in 96‐well plates containing primer mixtures. A total of 300 blood samples from unrelated volunteer donors in Taiwan were included in the study.

RESULTS
: All primers had specific amplification products. The typing results were available within 4 hours each time for up to four blood samples tested. Among the 13 HPAs, HPA‐3 had the greatest heterozygosity with a gene frequency of 0.3267, 0.4967, and 0.1767 for
HPA‐3a/HPA‐3a, HPA‐3a/HPA‐3b,
and
HPA‐3b/HPA3‐b,
respectively. For the remaining 12 HPAs, the predominance of
a/a
homozygosity was noted for HPA‐1, ‐2, ‐4, ‐5, and ‐6, with a frequency ranging from 0.9200 to 0.9967. The frequency of
a/a
homozygosity was 1.0000 for HPA‐7w to ‐13w, except for HPA‐10w, for which one case was observed to be
HPA‐10aw/HPA‐10bw
heterozygous. Excluding HPA‐3,
b/b
homozygosity was noted in only one case (
HPA‐6b/HPA‐6b ). The prevalence rates of HPA‐1 to ‐13w in this study were consistent with previous reports using different methods.

CONCLUSION : An extended, streamlined PCR‐SSP protocol for simultaneous genotyping of HPA‐1 to HPA‐13w was established. This allows fast and reliable diagnosis of alloimmune thrombocytopenia, and is readily applicable to large‐scale genetic population studies.
PEN110 treatment functionally inactivates the PBMNCs present in RBC units: comparison to the effects of exposure to gamma irradiation
Authors: Fast Loren D. , DiLeone Gilbert , Edson Clark M. , Purmal Andrei
Published: March 2003

BACKGROUND : The presence of WBCs in blood components is the major factor influencing the immunologic consequences of transfusion. Attempts to ameliorate these responses have used WBC reduction or inactivation by ionizing radiation. PEN110 (Inactine, V. I. Technologies) is a chemical that inhibits the replication of infectious pathogens by modifying their nucleic acids. These experiments compared effects of PEN110 treatment or gamma irradiation on WBC function.

STUDY DESIGN AND METHODS : Aliquots of non‐WBC‐reduced RBC units were treated with PEN110 or gamma irradiation with appropriate controls, and PBMNCs from these units were tested with in vitro assays. The assays included immunophenotyping, activating T cells with phorbol ester, proliferation of cells in response to mitogens or allogeneic cells, and determining the ability of cells to stimulate proliferative responses and to produce IL‐8. DNA fragmentation following PEN110 treatment was examined by PCR and terminal deoxynucleotidyl transferase‐mediated deoxyuridine triphosphate‐biotin nick end labeling assays.

RESULTS : Treatment of non‐WBC‐reduced RBC units with PEN110 functionally inactivated WBC in all in vitro assays used. In contrast, while gamma irradiation inhibited proliferation of the WBCs, it did not or only partially inhibited the ability of WBC to function in the other assays. PEN110, but not gamma irradiation, rapidly induced fragmentation of cellular DNA.

CONCLUSION : Because PEN110 was as effective as gamma irradiation at inhibiting WBC proliferation, PEN110 treatment could potentially be used to prevent the development of GVHD following transfusion.
Autologous transfusion recovery of WBC‐reduced high‐concentration platelet concentrates
Authors: Dumont Larry J. , Beddard Rachel , Whitley Pamela , Johnson Adrienne , McNeil Deanna , Sawyer Sherrie , Elfath M. Dean
Published: March 2003

BACKGROUND : This study evaluates the recovery and survival of high‐concentration platelets (HCPs) compared to standard apheresis platelets (APCs) in a double‐label autologous human system.

METHODS
: Nine HCP units paired with APC units were stored, labeled with either
51
Cr and
111 In, and returned, and recovery and survival were determined. Standard in vitro platelet biochemical and functional parameters were monitored over the storage period and evaluated in a secondary analysis.

RESULTS
: Three each HCP units containing more than 2.2 × 10
11
, 1.5 × 10
11
to 2.1 × 10
11
, and 0.8 × 10
11
to 1.1 × 10
11
platelets in 59.4 ± 2.5 mL were stored for 1, 2, or 5 days, respectively, and simultaneously with matched APC units (3.8 × 10
11 platelets, 282 mL). Recoveries were 72.3 ± 8.6, 60.8 ± 14.6, and 52.5 ± 6.7 percent for HCPs, respectively; and 59.4 ± 6.4 percent for APCs (p = 0.37). HCP survivals were 202.0 ± 14.9, 204.9 ± 10.2, and 162.6 ± 17.0 hours; APC survivals were 155.4 ± 20.3 hours (p = 0.001). Secondary analysis with P‐selectin added as a predictor in the model resulted in significant difference in recoveries for Day 1 HCPs versus Day 5 APCs (p = 0.024) with no difference shown for HCPs on Days 2 or 5 versus APCs. No significant difference was found in survival (p = 0.16).

CONCLUSION
: HCPs may be stored 24 hours for high yield, 48 hours for intermediate yield, and up to 5 days for yields less than 1.6 × 10
11 platelets per bag with equivalent to superior recovery and survival of platelets in the autologous transfusion model compared to APCs.
RBC‐associated IgG in patients with visceral leishmaniasis (kala‐azar): a prospective analysis
Authors: Vilela Rosana B. , Bordin José O. , Chiba Akemi K. , Castelo Adauto , Barbosa Maria C.
Published: March 2003

BACKGROUND: Despite the fact that anemia is one of the most striking clinical features of visceral leishmaniasis (kala‐azar), the factors involved in its pathogenesis are not fully understood. Although the cause of the anemia seen in these patients is often multifactorial, sequestration and destruction of the RBCs in the enlarged spleen, immune mechanisms, and alterations in RBC membrane permeability have been implicated.

STUDY DESIGN AND METHODS: To investigate whether RBCs of patients with kala‐azar were coated with IgG, blood samples of 67 patients were tested, prospectively, before (Day 1), during (Day 30), and after (Day 90) antimonial therapy, to examine the presence of RBC‐associated IgG, circulating immune complexes (CICs), and rheumatoid factor (RF).

RESULTS: The prevalence of a positive DAT on Day 1 was significantly greater than the prevalence of a positive DAT performed on Day 30 and on Day 90 (32.8 vs. 4 vs. 0%, p < 0.001). With an enzyme‐linked antiglobulin test (ELAT) to measure the number of IgG molecules per RBC more accurately, it was found that the amount of IgG molecules per RBC was increased (mean, 298 molecules of IgG per RBC) in the group of patients with kala‐azar tested before antimonial therapy, but was considered normal (<50 molecules of IgG per RBC) in all patients tested 90 days after treatment. The prevalence of a positive eluate test was low (15.0%) in DAT (anti‐IgG)‐positive patients and the positivity of DATs and ELATs correlated with the presence of either RF or CICs, respectively.

CONCLUSIONS: These data suggest that a nonspecific adsorption of CICs on the RBC surface is probably the most important factor involved in the increased amount of RBC‐associated IgG in patients with untreated visceral leishmaniasis; however, further prospective studies are required to establish the exact role of the RBC‐associated antibodies, CICs, and RF as contributing factors of the anemia seen in these patients.
IgG1 and IgG3 anti‐D in maternal serum and on the RBCs of infants suffering from HDN: relationship with the severity of the disease
Authors: Lambin Patrick , Debbia Martine , Puillandre Philippe , Brossard Yves
Published: March 2003

BACKGROUND : Anti‐D IgG antibodies that are responsible for severe cases of HDN belong chiefly to IgG1 and IgG3 subclasses. The relationship between the concentrations of IgG1 anti‐D and IgG3 anti‐D in maternal serum and the amount bound to the surface of infants' RBCs is not known. In addition, the contribution of the two subclasses to the severity of HDN is not well established.

STUDY DESIGN AND METHODS : Blood samples from 40 infants suffering from severe forms of HDN due to anti‐D were collected before transfusion together with sera from their respective mother. The amount of total anti‐D IgG as well as IgG1 anti‐D and IgG3 anti‐D on infants' RBCs and the concentration in maternal sera were determined by ELISA.

RESULTS : The median percentages of IgG1 anti‐D and of IgG3 anti‐D in maternal sera were 90 and 10 percent, respectively, whereas on infants' RBCs they were 97 and 3 percent, respectively. The differences between maternal and infantile percentages were significant (p < 0.001). IgG1 and IgG3 anti‐D bound to infants' RBCs increased concomitantly with the concentration of IgG1 and IgG3 anti‐D in maternal sera. The severity of HDN correlated positively with the concentration of IgG1 anti‐D in maternal sera, but negatively with the amount of IgG3 anti‐D bound to infants' RBCs. In addition, the existence of a high proportion of IgG3 anti‐D in maternal serum was associated with a delayed risk of fetal anemia.

CONCLUSION : The proportion of IgG3 anti‐D relative to the total anti‐D IgG on infants' RBCs is only one‐ third of the proportion present in maternal serum. The study of the correlations between the amount of IgG1 anti‐D and IgG3 anti‐D and the severity of HDN suggests that IgG1 anti‐D are more important than IgG3 anti‐D in the pathogenesis of fetal anemia.