bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro and in vivo persistence of reticulocytes from donor red cells
Authors: Perry E.S. , Moore R.H. , Berger T.A. , Billups L.C. , Maybee D.A. , Salata K.F. , Lippert L.E.
Published: March 2003

Background: Reticulocytes are important in the phenotyping of transfused patients. Reticulocytes can persist in blood units for the shelf life of the unit.

Study Design and Methods: Temperature dependence of reticulocyte persistence was examined in vitro at 4, 24, and 37°C by using thiazole orange staining and flow cytometric analysis. Two‐color flow cytometric analysis was used to evaluate the persistence of donor reticulocytes in transfused patients.

Results: Flow cytometric analysis using thiazole orange demonstrated that persistence of reticulocytes in units of stored CPDA‐1 blood was temperature‐dependent. Reticulocytes disappeared over 13 and 6 days at 24°C and 37°C, respectively, but at 4°C the reticulocyte count changed little over 35 days. Two‐color flow cytometric analysis of reticulocyte antigens was used to follow donor reticulocytes in 14 transfusion events in nine different patients. Donor reticulocytes persisted through 24 hours in 75 percent of the patients and were detectable at 48 hours in three patients.

Conclusion: This study demonstrates that reticulocytes persist during refrigerated storage; they are detectable in the circulation of most recipients for the first 24 hours after transfusion and in the circulation of a few recipients after 48 hours. These findings may have relevance for separation techniques based on reticulocyte density in samples drawn shortly after transfusion and for evaluation of reticulocyte counts in patients with hematologic abnormalities.
Anaphylaxis after treatment with recombinant factor VIII
Authors: Shopnick R.I. , Kazemi M. , Brettler D.B. , Buckwalter C. , Yang L. , Bray G. , Gomperts E.D.
Published: March 2003

Background: Treatment of hemophilia patients with recombinant factor VIII concentrates has not previously been associated with anaphylaxis.

Study Design and Methods: A 5‐week‐old boy with severe hemophilia A developed dyspnea, cyanosis, hypotension, and a diffuse urticarial rash following treatment with a recombinant factor VIII (Recombinate). To identify the cause of anaphylaxis in this patient, the vial lot was examined for the presence of endotoxin, and a checkerboard immunoblotting technique was used to test serum and/or plasma samples from the patient and mother for the presence of antibodies (IgA, IgG, IgE, and IgM) to Recombinate‐related antigens (recombinant factor VIII, von Willebrand factor, human serum albumin, Chinese hamster ovary proteins, bovine serum albumin, mouse monoclonal anti‐human factor VIII, polyethylene glycol 3350), and to ethylene oxide, the agent used to sterilize the infusion equipment.

Results: No immune response directed against the Recombinate‐related antigens or ethylene oxide that could be associated with the anaphylactic reaction was identified. Endotoxin was not present upon rabbit pyrogen testing of the therapeutic product.

Conclusion: These studies failed to show any association between Recombinate and the onset of the allergic reaction. This seems to be the first reported case of anaphylaxis following the infusion of a recombinant form of factor VIII concentrate.
Changes in blood counts after the administration of granulocyte‐colony‐ stimulating factor and the collection of peripheral blood stem cells from healthy donors
Authors: Stroncek D.F. , Clay M. E. , Smith J. , Ilstrup S. , Oldham F. , McCullough J.
Published: March 2003

BACKGROUND: After the collection of granulocyte‐colony‐stimulating factor (G‐CSF)‐mobilized peripheral blood stem cells from healthy donors, the donor platelet counts fall. However, the magnitude and duration of this decrease are not known. STUDY DESIGN AND METHODS: Sixty healthy people were given G‐CSF (5, 7.5, or 10 micrograms/kg/day) for 5 days (Days 1–5), and 1 peripheral blood stem cell component was collected on Day 6. The platelet count, white cell count, absolute neutrophil count, hematocrit, and red cell count were measured before administration of G‐CSF (Day 0), before collection of peripheral blood stem cells on Day 6, and on Days 8, 10, 13, 16, and 20.

RESULTS: The platelet count fell from 261 +/− 47 × 10(9) cells per L on Day 0 to 159 +/− 30 × 10(9) cells per L on Day 8 (p < 0.0001) and reached its lowest level on Day 10 (146 +/− 30 × 10(9)/L; p < 0.001). Compared to Day 0 levels, the platelet count was lower on Day 13 (185 +/− 49 × 10(9)/L, p < 0.001), was the same on Day 16 (270 +/− 53 × 10(9)/L), and was greater on Day 20 (333 +/− 60 × 10(9)/L, p < 0.0001). The white cell count returned to pretreatment values on Day 13, and the absolute neutrophil count returned to pretreatment values on Day 10 (Day 0 white cell count = 6.05 +/− 1.59 × 10(9)/L and Day 0 absolute neutrophil count = 3.97 +/− 1.52 × 10(9)/L). On Day 20, both were less than pretreatment values (white cell count = 5.14 +/− 1.24 × 10(9)/L, p = 0.0007 and absolute neutrophil count = 3.20 +/− 1.24 × 10(9)/L, p = 0.0036). The red cell counts on Day 16 (4.52 +/− 0.41 × 10(12)/L) and Day 20 (4.42 +/− 0.39 × 10(12)/L) were less than Day 0 values (4.73 +/− 0.43 × 10(12)/L, p = 0.008 and p < 0.0001, respectively). The hematocrit on Day 20 (39.2 +/− 3.2%) was also less than that on Day 0 (41.2 +/− 4.8%; p = 0.01). The changes in these blood counts were not affected by the dose of the G‐CSF.

CONCLUSION: After stimulation with granulocyte‐colony‐stimulating factor and the collection or peripheral blood stem cells, the platelet counts in normal donors were decreased for at least 7 days (Days 6–13). Two weeks after collection of peripheral blood stem cells (Day 20), platelet production was increased, but the production of neutrophils and red cells was decreased. If two or more peripheral blood stem cell components are collected, then the platelet count should be measured after the second and subsequent collections. Further studies on the long‐term effect of G‐CSF on blood counts are needed.
Expression and quantitative variation of the low‐incidence blood group antigen He on some S‐s‐red cells
Authors: Reid M. E. , Storry J. R. , Ralph H. , Blumenfeld O. O. , Huang C.‐H.
Published: March 2003

BACKGROUND: Red cells devoid of glycophorin B (GPB)‐borne S, s, and U antigens are classified as an S‐s‐U‐ or S‐s‐U variant (U+var) and can arise from deletion and nondeletion genetic backgrounds. In nondeletion forms of S‐s‐U‐, little information is available on whether the altered GPB gene (GYPB) is expressed in red cells. STUDY DESIGN AND METHODS: Red cells classified as S‐s‐U‐ or S‐s‐U+var were tested with anti‐U, anti‐U/GPB, anti‐He, and anti‐N by hemagglutination. Selected samples were tested by flow cytometry, immunoblotting, and polymerase chain reaction amplification using allele‐specific primers. RESULTS: He (MNS6) was found on 23 percent (20/87) of samples. These and another 21 of the 87 samples were agglutinated by an anti‐U/GPB reagent; this indicated that approximately 50 percent of S‐s‐samples possessed GPB variants. The strength of He varied among the samples. Genomic polymerase chain reaction with allele‐specific primers showed the presence of expected DNA GPB‐like products encoding He. Immunoblotting showed that He was carried on a membrane component with a relative molecular mass indistinguishable from that of GPB. CONCLUSION: The finding of He on S‐s‐ red cells provides direct evidence for the presence of an altered form of GPB in red cells previously thought to be devoid of this glycophorin. Quantitative variation in He antigen expression was observed in a subset of S‐s‐ red cells.
A simple and rapid competitive enzyme‐linked immunosorbent assay to identify HPA‐1a (PlA1)‐negative donor platelet units
Authors: Denomme G. , Horsewood P. , Xu W. , Smith J. , Kelton J.
Published: March 2003

Background: Alloantibodies to HPA‐1a (PlA1) are the major cause of neonatal alloimmune thrombocytopenia and posttransfusion purpura and have been implicated in refractoriness to random‐donor platelet transfusions. However, most assays used to phenotype platelets are cumbersome or time‐consuming for large numbers of samples.

Study Design and Methods: A simple, competitive (inhibition) enzyme‐linked immunosorbent assay for HPA‐1a phenotyping of donor platelets was developed. A segment from the donor platelet unit transfer line was sealed to obtain a small aliquot of platelets. These platelets were washed once and added to a predetermined dilution of serum containing alloantibodies to HPA‐1a. Residual anti‐HPA‐1a binding to the glycoprotein IIb/IIIa purified by lectin and high‐performance liquid chromatography and coated on microtiter wells was detected with a conjugated antihuman IgG. A lack of inhibition equivalent to control (no platelets) was used to determine that the platelets were HPA‐1b/b.

Results: Of the 557 platelet units tested, 14 (2.5%) were found to be HPA‐1a negative, and they were confirmed to be HPA‐1b/b by DNA genotyping. Two of the 14 HPA‐1b/b units were also HPA‐3b/b (approx. 0.35% of the random population). Use of the microtiter format allows 100 to 200 samples to be processed per day.

Conclusion: This simple and inexpensive assay is useful for identifying HPA‐1b/b units for platelet‐compatible transfusions or for platelet antibody investigations.
Effectiveness of very low doses of subcutaneous recombinant human erythropoietin in facilitating autologous blood donation before orthopedic surgery
Authors: Sans T. , Bofill C. , Joven J. , Clivillé X. , Simó J.M. , Llobet X. , Peró A. , Galbany J.
Published: March 2003

Background: Clinical and pharmacokinetic data suggest that very low doses of subcutaneous recombinant human erythropoietin (rHuEPO) may be effective in a preoperative autologous blood deposit program.

Study Design and Methods: Fifty‐two patients, scheduled for orthopedic surgery, were enrolled in a double‐blind and placebo‐controlled study. Patients were randomly assigned to the placebo group or to receive 30, 60, or 100 IU per kg of rHuEPO subcutaneously twice a week for 2 weeks before surgery. The dose of rHuEPO that was effective in facilitating the collection of 4 units of blood in the 2 weeks before surgery and that prevented a sharp decrease in hematocrit was determined.

Results: Only in patients receiving 100 IU per kg of rHuEPO did the outcome measurements differ significantly from those in the placebo group. With a higher (p < 0.01) cumulative increase in red cell volume during the study period (297 ± 127 vs. 121 ± 44 mL), 64 percent of those receiving 100 IU per kg of rHuEPO were able to donate 4 units of blood for autologous use, as compared with 23 percent of the placebo group (p < 0.05). Allogeneic transfusion was avoided, and the preoperative hematocrit and reticulocyte count were significantly higher in the patients receiving 100 IU per kg of rHuEPO (p < 0.05 and p < 0.01, respectively).

Conclusion: Subcutaneously administered rHuEPO at a dose of 100 IU per kg twice a week for 2 weeks is effective in facilitating the collection of blood for autologous use and may improve the cost‐benefit ratio of blood conservation interventions. Doses ≤ 60 IU per kg are ineffective in facilitating such collections in this surgical setting.
Potential increased risk of virus transmission due to exclusion of older donors because of concern over Creutzfeldt‐Jakob disease
Authors: Busch M.P. , Glynn S. A. , Schreiber G. B.
Published: March 2003

BACKGROUND: Concern over the theoretical possibility of disease transmission via blood from donors who develop Creutzfeldt‐Jakob disease has led to proposals to exclude older individuals from donating plasma for further manufacture into pooled plasma donations. The impact of extending this age‐deferral policy to blood donors was examined with respect to the risk for known transmissible viruses. STUDY DESIGN AND METHODS: Demographic characteristics and confirmed prevalence rates (/10(5) first‐time donations) and incidence rates (/10(5) person‐years for repeat donors) for viral markers were compared for donors < 50 years old (n = 1,259,805 [85%]) and > or = 50 years old (n = 219,856 [15%]) and for donors < 60 years old (n = 1,409,176 [95%]) and > or = 60 years old (n = 70,485 [5%]). Incidence rates were combined with infectious window‐period estimates for each virus, to calculate the risk of virus transmission per 10(6) donations. RESULTS: Unadjusted prevalence rates were significantly greater for younger than for older donor groups for human immunodeficiency virus (HIV), hepatitis B surface antigen (HBsAg), and hepatitis C virus (HCV) (p < or = 0.05). Incidence rates (and transmission risk estimates) for HBsAg were significantly higher in the < 50 donor group than in the > or = 50 group (p < or = 0.05), and those for HIV, human T‐lymphotropic virus, and HCV were not significantly higher (p > 0.05). Blanket removal of donors over the age of 50 would potentially lead to the following significant increases in the risk of infected units: HIV, 12 percent; HCV, 21 percent; and hepatitis B virus (HBsAg), 22 percent. CONCLUSION: Removal of donors over the age of 60 would not significantly affect the risk of infected units. Deferral of donors > or = 50 years of age from whole‐blood donations for unfounded concerns about Creutzfeldt‐Jakob disease could have adverse effects on both blood availability and safety.
Inhibition of cytokine accumulation and bacterial growth during storage of platelet concentrates at 4° C with retention of in vitro functional activity
Authors: Currie L.M. , Harper J.R. , Allan H. , Connor J.
Published: March 2003

BACKGROUND: The potential for bacterial contamination limits the storage of platelet concentrates (PCs) at 22° C to 5 days. In addition, storage of platelets under conventional protocols for longer times (> 3 days), in the absence of white cell filtration, has been correlated with incidents of cytokine‐associated febrile reaction in recipients. It has been demonstrated that the addition of a reagent mixture of second‐messenger effectors allows platelets stored at 4°C to maintain significant in vitro functional activity. Thus, the effects of 4°C storage on the growth of bacteria and the accumulation of cytokines by the white cell fraction of PCs were analyzed to demonstrate the benefits of this refrigerated storage system.

STUDY DESIGN AND METHODS: The platelet storage solution was added directly to PCs obtained from the blood bank, and these treated PCs were stored at 4° C without agitation. In parallel, control PCs were stored according to standard blood‐banking procedures. On Days 1, 3, 5, and 9, the PCs were measured for the plasma concentrations of cytokines. Treated and control PCs stored at 4°C and 22°C were inoculated with low‐titer Staphylococcus aureus, and bacterial growth was measured over a 5‐day period.

RESULTS: Control PCs displayed a time‐dependent increase in the plasma concentration of interleukin 6, interleukin 1β, and tumor necrosis factor α. These conventionally stored PCs also displayed a time‐dependent increase in the bacteria titer. In contrast, the treated PCs stored at 4°C displayed no accumulation of the above cytokines in the plasma fraction and no increase in bacteria titer above the initial inoculation.

CONCLUSION: The storage of PCs at refrigerated temperatures inhibits the accumulation of white cell‐produced cytokines in the PCs, an effect that could alleviate cytokine‐associated febrile transfusion reactions The 4°C storage was also bacteriostatic, which indicates that the storage of PCs at that temperature increases safety by decreasing the potential for sepsis. Thus, the ability to store PCs at 4°C may allow extension of the storage limit beyond 5 days.
A moderate transfusion regimen may reduce iron loading in beta‐ thalassemia major without producing excessive expansion of erythropoiesis
Authors: Cazzola M. , Borgna‐Pignatti C. , Locatelli F. , Ponchio L. , Beguin Y. , De Stefano P.
Published: March 2003

BACKGROUND: Hypertransfusion with a baseline hemoglobin of 10 to 12 g per dL is still considered by many to be the mainstay of conservative therapy for beta‐thalassemia major. However, this regimen is frequently associated with manifestations of transfusion iron overload, despite regular chelation therapy with subcutaneous desferoxamine. STUDY DESIGN AND METHODS: To verify whether a transfusion regimen with a target pretransfusion hemoglobin level between 9 and 10 g per dL can allow a significant reduction in blood consumption, while still effectively suppressing erythropoiesis, the records were reviewed of 32 beta‐ thalassemia major patients, who were maintained at a pretransfusion hemoglobin of 11.3 +/− 0.5 g per dL between 1981 and 1986. These patients were switched at the beginning of 1987 to a transfusion regimen with pretransfusion hemoglobin of 9.4 +/− 0.4 g per dL. The degree of erythroid marrow activity was evaluated in these patients and in 32 subjects with beta‐thalassemia intermedia through the simple measurement of serum transferrin receptor. RESULTS: After the adoption of the moderate transfusion regimen, transfusion requirements decreased from 137 +/− 26 to 104 +/− 23 mL per kg per year of red cells (p < 0.0001), and mean serum ferritin decreased from 2448 +/− 1515 to 1187 +/− 816 micrograms per L (p < 0.0001), with one‐half of patients achieving serum ferritin levels lower than 1000 micrograms per L. The proportion of patients having spontaneous pubertal development increased significantly (p < 0.01), as a result of less iron‐related gonadotropin insufficiency. At the lower pretransfusion hemoglobin, erythroid marrow activity did not exceed two to three times normal levels in most subjects. CONCLUSION: As compared with hypertransfusion, moderate transfusion may allow more effective prevention of iron loading, with higher likelihood of spontaneous pubertal development and without producing excessive expansion of erythropoiesis.
A retrospective analysis of microbial contaminants in outdated random‐donor platelets from multiple sites
Authors: Leiby DA. , Kerr K.L. , Campos J.M. , Dodd R.Y.
Published: March 2003

BACKGROUND: Platelet components contaminated with bacteria are an important source of transfusion‐associated bacterial sepsis. Estimates of contamination rates vary widely (0–10%) and are highly controversial. The present study, designed with stringent testing regimens, retrospectively determined the prevalence of microbial contaminants in platelets from four collection regions.

STUDY DESIGN AND METHODS: During a 9‐month period, outdated platelet units were assayed by spreading aliquots from the unit, and from thioglycollate broth medium inoculated with part of the unit, onto 5‐percent sheep blood agar media. Cultures were examined after 72‐hour incubation at 37°C, and, if bacterial growth was present, the assay processes were repeated with fresh inocula. Units were considered contaminated only if repeatedly positive.

RESULTS: Four (0.08%) of 4995 units sampled were contaminated, two with Corynebacterium sp. and one each with Propionibacterium acnes and Aspergillus terreus. Contaminants were present at low, subclinical levels and were detected only after amplification in thioglycollate. The contaminated units were cultured 1, 2, 3, and 7 days after expiration.

CONCLUSION: Contamination rates were low and did not vary by region. The identification of A. terreus suggests the role that transfusion may play in transmitting fungal infections should be reassessed. The persistent detection of contaminated platelet units supports the need for a test to detect clinically relevant levels of microbial contaminants in blood components.
Anti‐D in a D‐positive renal transplant patient
Authors: Saba N.F. , Sweeney J.D. , Perm L.C. , Lawton J.C. , Yankee R.L. , Huang C.H. , Schanfield M.S.
Published: March 2003

BACKGROUND: The detection of anti‐D in a D‐positive renal transplant recipient is unusual and may arise by several potential mechanisms. These include passive transfer of alloantibody and the presence of autoanti‐D or alloanti‐D that is due to microchimerism when the allograft is from a D‐negative donor. In the latter case, overt hemolysis has been seen or suspected. The occurrence of anti‐D in a D‐positive renal transplant recipient without hemolysis, which is most likely attributable to microchimerism, is reported.

CASE REPORT: A 51‐year‐old group O, D‐positive woman, who was serologically HLA type A1, A2; B8, B44; DR3, DR6, DR52; DQ1, DQ2, underwent the transplantation of a kidney from a cadaveric donor who was serologically HLA type A1, A2; B8, B44; DR13, DR17, DR52; DQ1, DQ2. The donor was known to be D‐negative and immunized to D. No Wood components or derivatives were administered at the time of organ graft. Ten weeks after the transplant, the direct antiglobulin test was positive in the recipient, and anti‐D was eluted. Polymerase chain reaction amplification using primers to distinguish DR13 (donor) from DR14 alleles (recipient split of DR6) in the peripheral Wood showed the recipient to be DR14. No DR13 could be detected, and thus microchimerism could not be confirmed. However, in the peripheral blood, GM and KM allotyping of the serum (GM A,F,X B,G and KM 1,3) and eluate (G1M F, KM 3) showed a pattern of allotypes most consistent with an alloantibody. Eleven months after transplantation, the graft continued to function; the direct antiglobulin test was still positive, and elution of anti‐D persisted.

CONCLUSION: This case of anti‐D in a D‐positive renal transplant recipient is attributed to microchimerism, despite the lack of confirmation by genotypic analysis of the peripheral Wood. It raises the possibility that microchimerism may be a more common phenomenon in solid allograft recipients than is realized.
Screening with monoclonal anti‐Fy3 to provide blood for phenotype‐ matched transfusions for patients with sickle cell disease
Authors: Sandler S. G. , Mallory D. A. , Wolfe J. S. , Byrne P. C. , Lucas D. M.
Published: March 2003

BACKGROUND: In the United States, there is a shortage of blood group phenotype‐matched red cells (RBCs) for patients with sickle cell disease (SCD). A protocol designed to supply phenotype‐matched RBCs for these patients by combining the recruitment of African American blood donors and automated testing of RBCs for these patients for the presumptive Fy(a‐b‐) phenotype using monoclonal anti‐Fy3 was evaluated. STUDY DESIGN AND METHODS: African American donors were recruited, to increase the likelihood of phenotype matches in the donor population. Samples of RBCs were tested for the presumptive Fy(a‐b‐) phenotype by using monoclonal anti‐Fy3 and an automated blood typing analyzer. RBCs confirmed to be Fy(a‐b‐) were retyped for selected Rh, MNS, Kell, Duffy, and Kidd blood system antigens. The extended phenotypes were matched with those of 41 SCD patients requiring transfusions. RESULTS: Of 8323 blood donations during the study, approximately 40 percent (3329) were made by African Americans. Approximately 22 percent (737) of African Americans were identified as Fy(a‐b‐) by this protocol and 12 percent (410) were phenotype matches for the 41 SCD patients. CONCLUSION: Combining the recruitment of African American blood donors and automated phenotyping using monoclonal anti‐Fy3 offers a practical, relatively low‐cost strategy for supplying phenotype‐matched RBCs for SCD patients. This protocol increases the options for addressing the shortage of phenotype‐matched RBCs for SCD patients.
Differentiation of anti‐D, ‐C, and ‐G: clinical relevance in alloimmunized pregnancies
Authors: Shirey R.S. , Mirabella D.C. , Lumadue J.A. , Ness P.M.
Published: March 2003

BACKGROUND: The differentiation of anti‐D, ‐C, and ‐G specificities is seldom considered clinically important in pretransfusion testing. However, distinguishing these antibody specificities in alloimmunized pregnancies may be essential. The clinical prognosis as well as Rh immune globulin prophylaxis depends on the accurate identification of these antibodies.

CASE REPORT: A pregnant woman, para 1 gravida 4, who had received Rh immune globulin at appropriate intervals during her previous pregnancies was reported to have anti‐D (titer = 4) and anti‐C (titer = 32). Differential adsorption and elution studies showed that the patient had anti‐C and anti‐G, but not anti‐D. This case prompted retrospective examination of the sera from six other women with anti‐D and anti‐C who were referred to a high‐risk pregnancy clinic. Of six pregnant women reported to have anti‐D and anti‐C; two had anti‐D, ‐C, and ‐G; three had anti‐D and ‐G, but not anti‐C; and one had anti‐C and ‐G, but not anti‐D. This last is similar to the index case.

CONCLUSION: Cases of pregnant women with anti‐C and ‐G, but not anti‐D, are not infrequent. Studies to differentiate anti‐D, ‐C, and ‐G should be performed on alloimmunized pregnant women presumptively identified as having anti‐D and anti‐C when the medical history (Rh immune globulin prophylactic therapy) and/or titer values (e.g., anti‐C titer higher than anti‐D titer) suggest that anti‐D may not actually be present. Rh immune globulin has not failed in these patients, and they should receive this therapy during pregnancy to prevent immunization to D.
Transfusions of granulocyte‐colony‐stimulating factor‐mobilized granulocyte components to allogeneic transplant recipients: analysis of kinetics and factors determining posttransfusion neutrophil and platelet counts
Authors: Adkins Douglas R. , Spitzer Gary , Johnston Marilyn , Velasquez William S. , Dunphy Frank R. , Petruska Paul J.
Published: March 2003

BACKGROUND: Granulocyte‐colony‐stimulating factor (G‐CSF) is a safe and effective agent for mobilization of neutrophils in normal donors, consistently resulting in cell yields per leukapheresis (LA) procedure that are superior to those with other agents. LA components also contain platelets, whose clinical relevance is unknown. STUDY DESIGN AND METHODS: This study describes the kinetics of and analyzes the factors determining the ANC and platelet count increments seen with each of three transfusions of granulocytes collected from HLA‐matched sibling donors receiving G‐CSF (n=10; maximum of 3 LA procedures/donor). The transfusions were given to recipients (n=10) on alternate days beginning. Day 1 after allogeneic bone marrow transplant (BMT). RESULTS: Significant, sustained increments in the recipient ANCs were observed after the transfusion of G‐CSF‐mobilized LA components. The mean peak posttransfusion increments in the ANCs were 1195, 729, and 631 per microL with transfusion of donor LA components on Days 1, 3, and 5, respectively. The length of time that the mean posttransfusion ANC was at or above the baseline (pretransfusion) value was 25 to 37 hours, depending on the post‐BMT day when the component was administered. No consistent relationship was observed between LA component granulocyte dose, baseline recipient ANC, or temperature elevation and post‐transfusion ANC increments. Large numbers of platelets (mean, 2.55 × 10(11)) were present in LA components, and this resulted in significant increments from baseline in the mean platelet count 1 hour after LA component transfusions. Between Days 1 and 7, the duration of severe neutropenia was shorter and the percentage of patients requiring nondonor platelet transfusions was less in study patients who received LA component transfusions than in a similar historical control group who did not. CONCLUSION: The transfusion of G‐CSF‐mobilized, HLA‐matched LA components to allogeneic BMT recipients resulted in significant and sustained increments in the ANC and the platelet count. Within the range examined, a relationship between neutrophil dose and an increment in the ANC was not demonstrated.
Leukocyte phenotypic changes in an in vitro model of ABO hemolytic transfusion reaction
Authors: Udani M. , Rao N. , Telen M. J.
Published: March 2003

BACKGROUND: ABO antigen‐antibody interaction in the presence of peripheral blood leukocytes (white cells) results in the production of a variety of proinflammatory cytokines. However, although tumor necrosis factor alpha has been shown to be derived at least primarily from monocytes, the range of cells activated by this process has not previously been reported. Therefore, changes in mononuclear cell surface antigen expression were studied, to determine which subsets of white cells appeared to be activated in the setting of ABO incompatibility. STUDY DESIGN AND METHODS: Group O peripheral blood mononuclear cells (PBMCs) were incubated in autologous plasma with group A or O red cells (RBCs) for up to 24 hours. White cell expression of activation and adhesion markers was measured at 2 and 24 hours by flow cytometry, using direct or indirect fluorescein or phycoerythrin labeling. RESULTS: Expression of lymphocyte activation markers CD25, CDw108, and CD109 was equivalent when PBMCs incubated with group A and O RBCs were compared. However, after 2 hours, mean fluorescence of CD14 on PBMCs incubated with group A RBCs was 65 percent of that on PBMCs incubated with group O RBCs and remained similarly decreased at 24 hours. CD44 expression was upregulated on PBMCs exposed to both group A and O RBCs, but it was increased significantly more on monocytes exposed to group A RBCs. The ability to bind hyaluronic acid was induced in approximately 42 percent of CD14+ monocytes exposed to group A RBCs but in no cells exposed to group O RBCs. CONCLUSION: Downregulation of CD14 and increased binding of hyaluronic acid reflects monocyte activation in this model. No evidence of lymphocyte activation was found, supporting the hypothesis that ABO transfusion reactions primarily activate monocytes.
Prospective validation of a point score system for predicting blood transfusion following hip or knee replacement
Authors: Larcique B. J. , Gilbert K. , Brien W. F.
Published: March 2003

BACKGROUND: The purpose of this study was to validate a previously published point score system for predicting the likelihood of a postoperative blood transfusion following hip or knee replacement.

STUDY DESIGN AND METHODS: Data were collected prospectively on 460 sequential patients undergoing elective hip and knee replacement at two academic hospitals. Blood transfusion frequency was determined for patients in each of the four risk strata, as defined by the point score system. The accuracy of the system was validated by calculating the area under the receiver operating characteristic (ROC) curve for each site. Data were then combined and inappropriate blood transfusions were eliminated, by using the American College of Physicians guidelines. The frequencies of blood transfusion within each strata were recalculated along with an ROC curve.

RESULTS: The point score system accurately predicted the likelihood of blood transfusion at both hospitals, despite marked differences in overall transfusion frequencies. The calculated areas under the ROC curves were 0.78 and 0.79 for the two sites. The point score system also proved valid when only appropriate blood transfusions were considered, with a calculated area under the ROC curve of 0.74.

CONCLUSION: The point score system can accurately predict the likelihood of postoperative blood transfusion following hip or knee replacement. Such a system can be used to target high‐risk patients for preoperative autologous blood donation.
Sample suitability for the detection of minor white cell populations (microchimerism) by polymerase chain reaction
Authors: Reed W. , Lee T.‐H. , Vichinsky E.P. , Lubin B.H. , Busch M.P.
Published: March 2003

BACKGROUND: There is increasing use of highly sensitive testing with polymerase chain reaction (PCR) to study white cell microchimerism after transfusion and transplantation. This study investigated possible artifactual sources of allogeneic sample contamination before PCR testing. STUDY DESIGN AND METHODS: Quantitative Y‐chromosome PCR was used to study microchimerism among transfused patients with sickle cell disease (SCD) and thalassemia by using residual specimens from the clinical laboratory. High levels of circulating male white cells among transfused patients with SCD but not thalassemia led to concern over the artifactual origin of male cells. To investigate, paired specimens were collected from 26 female SCD patients: one specimen underwent processing only for PCR, while the other underwent testing in the clinical laboratory before PCR as a process control. All laboratory instruments were also assessed for their ability to impart male allogeneic cells to aliquots of female blood. RESULTS: Thirty‐three (31%) of 107 SCD samples, but 0 of 20 thalassemia samples, gave a high‐ level PCR signal. One of 26 paired samples that was not exposed to clinical laboratory equipment had low‐level PCR positivity while 10 of the 26 became strongly positive after testing on a blood cell analyzer and a reticulocyte analyzer. Sixteen of 32 female samples became positive after reticulocyte analysis, while none became positive after blood cell analysis. Samples from thalassemia patients tested PCR‐ negative because reticulocyte counts had not been performed. CONCLUSION: Allogeneic cell contamination is common with clinical laboratory equipment. These samples may not be suitable for microchimerism studies. In addition to method controls, process controls should be employed where appropriate.
Collection of allogeneic peripheral blood progenitor cells by two protocols on an apheresis system
Authors: Schreiner T. , Wiesneth M. , Krug E. , Kalinova J. , Büchele H. , Maccari B. , Erne E. , Bischof C. , De Reys S.
Published: March 2003

BACKGROUND: Granulocyte‐colony stimulating factor‐mobilized allogeneic peripheral blood progenitor cells (PBPCs) are replacing bone marrow in transplantation for the treatment of several hematologic malignancies. The advantages of PBPCs are offset by the donor‐associated disadvantages of granulocyte‐colony stimulating factor side effects and the risk of apheresis‐like platelet loss. STUDY DESIGN AND METHODS: For each individual, the first donation of allogeneic PBPCs by apheresis on the Spectra, using either the standard protocol Version 4.7 (45 donors, [Version 4.7]) or the AutoPBSC (60 donors, [AutoPBSC]) was compared. Between July 1995 and May 1996, all donors enrolled underwent Version 4.7 apheresis. Since May 1996, the majority of donors underwent AutoPBSC apheresis. For statistical analysis, only data from the first apheresis for each individual donor was considered for independent values. RESULTS: These results indicate a similar collection efficiency for CD34+ cells in the first apheresis of each donor (54% Version 4.7 vs. 53% AutoPBSC, p = 0.8). The apheresis time was longer with the AutoPBSC (233 min vs. 251 min, p = 0.005), whereas the loss of platelets was significantly lower (p < 0.001) with the AutoPBSC (28% vs. 19%). The mean number of CD34+ cells collected in the first apheresis component was 4.0 × 10(8) (Version 4.7) versus 3.8 × 10(8) (AutoPBSC). CONCLUSION: Both apheresis protocols collect sufficient numbers of PBSCs for allogeneic transplantation. The AutoPBSC operates in a fully automatic fashion, avoiding manual adjustment and interindividual variations. The loss of platelets is lower with AutoPBSC than with Version 4.7, but the apheresis time is slightly longer.
Adverse reactions associated with autologous blood transfusion: evaluation and incidence at a large academic hospital
Authors: Domen R.E.
Published: March 2003

BACKGROUND: It is acknowledged that autologous blood is the safest for the patient to receive. However, it is generally not appreciated that transfusion reactions to autologous blood may occur, despite the fact that it is the patient's own blood. STUDY DESIGN AND METHODS: A retrospective review of all transfusion reactions reported to a hospital transfusion service from 1991 through 1996 was performed, and all reactions to autologous blood were further investigated. RESULTS: Reported adverse reactions to autologous blood composed 2.1 percent of all transfusion reactions investigated in the hospital, involving 0.16 percent (15/9,353) of all transfused preoperatively donated autologous red cell units and 0.027 percent (5/18,506) of all intraoperatively salvaged units. Further investigation revealed that 60 percent (12/20) of these adverse reactions were felt to be clinically important and directly attributable to the autologous blood transfusion. Adverse reactions included febrile nonhemolytic (5) and allergic (4) reactions, an acute hemolytic transfusion reaction secondary to a clerical error (1 intraoperatively salvaged unit), and other nonsignificant adverse reactions (2). Eight adverse reactions were determined these reactions to be unrelated to the autologous transfusion. CONCLUSION: Despite the fact that the blood given is the patient's own blood, transfusion reactions to autologous blood do occur. As it is for allogeneic transfusion, any suspected adverse reaction to autologous blood transfusion should be investigated.
The Collaborative Hospital Transfusion Study: variations in use of autologous blood account for hospital differences in red cell use during primary hip and knee surgery
Authors: Churchill W. H. , McGurk S. , Chapman R. H. , Wallace E. L. , Bertholf M. F. , Goodnough L. T. , Kao K. J. , Olson J. D. , Woodson R. D. , Surgenor D. M.
Published: March 2003

BACKGROUND: Red cell use in patients undergoing Diagnosis Related Group (DRG) 209 procedures (major joint and limb reconstruction procedures of the lower extremities) has been shown to have large, unexplained interhospital variations.

STUDY DESIGN AND METHODS: Abstracted records of 2590 consecutive DRG 209 patients at five university hospitals from January 1992 to December 1993 were stratified by procedure and preoperative blood deposit status. Patient characteristics and transfusion and in‐hospital outcomes were compared across hospitals.

RESULTS: Blood use among patients who did not preoperatively deposit blood was similar across hospitals. Significant differences were found across hospitals for total hip replacement patients in the percentage of patients preoperatively depositing blood (59–80%), percentage of patients receiving transfusion(s) (51 to > 99%), the mean number of units collected per patient (1.6‐2.9), and the mean number of unused autologous units per 100 patients (1–185). No significant differences were found in the percentage of those who deposited blood and then required allogeneic units. There was little variability in length of hospital stay or in last hematocrits. Findings were similar for total knee replacement patients.

CONCLUSIONS: Interhospital variations in red cell use for primary total hip and knee reconstruction are primarily due to hospital‐specific differences in autologous blood collection and transfusion.
Survival of fresh human platelets in a rabbit model as traced by flow cytometry
Authors: Rothwell S. W. , Maglasang P. , Krishnamurti C.
Published: March 2003

BACKGROUND: In the event of hemorrhage and blood loss, platelets play a vital role in the coagulation process. However, there are currently no acceptable protocols for long‐term storage of platelets. As a first step toward testing the efficacy of stored platelets or platelet substitutes in vivo, a flow cytometric technique was developed to detect human platelets in rabbit blood.

STUDY DESIGN AND METHODS: Human platelets were transfused to rabbits whose reticuloendothelial system was inhibited by the administration of ethyl palmitate. Because human and rabbit platelets display surface molecules with different epitopes, human platelets were selectively labeled with antibodies specific for glycoprotein IX (CD42a). As this antibody does not label rabbit platelets, it allows discrimination of human from rabbit platelets in samples of rabbit blood containing both types of platelets.

RESULTS: Survival of human platelets in rabbits was monitored by flow cytometry and fluorescence microscopy in blood drawn at various times after the platelet transfusion. Fresh human platelets transfused to untreated control rabbits (n = 3) were removed from circulation within 10 minutes of the completion of the transfusion. Fresh platelets (1 day old) transfused to rabbits treated with ethyl palmitate (n = 5) survived for 24 hours with an average half‐life of 8.6 hours. In contrast, 8‐day‐old platelets were cleared from the circulation sooner with an average half‐ life of 2.9 hours (n = 4).

CONCLUSION: This report describes a rapid and efficient method of assessing the survival of human platelets in a rabbit model using flow cytometry. This technique will enable the monitoring in rabbits of human platelets prepared by various preservation protocols.
Hemochromatosis probands as blood donors
Authors: Barton J.C. , Grindon A.J. , Barton N.H. , Bertoli L.F.
Published: March 2003

BACKGROUND: There has been no estimate of the potential eligibility of hemochromatosis probands or patients as blood donors or the suitability for transfusion of their blood that was removed by therapeutic phlebotomy.

STUDY DESIGN AND METHODS: According to guidelines of the American Association of Blood Banks, a retrospective estimate of these factors in 211 adult white hemochromatosis probands diagnosed during routine medical care was performed. The findings were compared to those in volunteer white whole‐blood donors.

RESULTS: Before diagnosis of hemochromatosis, 49 probands had voluntarily donated 597 units of blood; 88 percent were donated by men. After diagnosis, 142 (67%) of 211 probands were potentially eligible. Data on each unit removed during iron‐depletion therapy and during the first year of maintenance therapy (therapeutic phlebotomy) were available in 86 eligible probands. Of 1592 units, 1029 (65%) obtained during iron‐depletion therapy in eligible probands were potentially suitable; 86 percent were from men. During maintenance therapy, 106 (88%) of 121 units from eligible probands were potentially suitable. In volunteer donors, 255,567 (94%) of 273,302 presenting donors were accepted. After testing and laboratory losses, 239,300 (94%) units were acceptable for transfusion.

CONCLUSIONS: In comparison with normal volunteers, hemochromatosis probands at diagnosis are less likely to be eligible as blood donors. The percentage of units obtained from patients during iron‐depletion therapy that are suitable for transfusion is also lower, although the percentage increases during maintenance therapy.
Platelet surface p‐selectin, platelet–granulocyte heterotypic aggregates, and plasma‐soluble p‐selectin during plateletpheresis
Authors: Barnard M.R. , MacGregor H. , Mercier R. , Ragno G. , Pivacek L.E. , Hechtman H.B. , Michelson A.D. , Valeri C.R.
Published: March 2003

BACKGROUND: Plateletpheresis components have been shown to contain p‐selectin‐positive platelets after collection and storage. P‐selectin mediates binding of activated platelets to granulocytes and monocytes. This study was undertaken to assess platelet activation, granulocyte activation, platelet–granulocyte heterotypic aggregate formation, and the plasma‐soluble p‐selectin level during plateletpheresis performed on a particular instrument (MCS+, Haemonetics).

STUDY DESIGN AND METHODS: Flow cytometry was used to assay platelet surface p‐selectin, granulocyte iC3b receptor, and platelet–granulocyte aggregates in the platelet component, residual blood in the disposable polycarbonate bowl of the MCS+, and in the donor blood with and without the addition of in vitro agonists before, during, and after plateletpheresis. The plasma‐soluble p‐selectin levels in the platelet component, disposable bowl, and donor venous blood were measured by an enzyme‐linked immunosorbent assay.

RESULTS: Levels of p‐selectin‐positive platelets, activated granulocytes, and platelet–granulocyte aggregates were greater in the disposable bowl than in the preapheresis donor blood. Levels of p‐selectin‐positive platelets, activated granulocytes, and platelet–granulocyte aggregates in the postapheresis donor blood were similar to those in the preapheresis donor blood. The platelet components contained no activated granulocytes or detectable platelet–granulocyte heterotypic aggregates, and only about 10‐percent activated platelets. The plasma‐soluble p‐selectin level in the platelet component was significantly greater than that in the preapheresis donor blood, the residual blood in the disposable bowl, or the postapheresis donor blood.

CONCLUSIONS: Measurements of platelet surface p‐selectin, platelet–granulocyte heterotypic aggregates, and plasma‐soluble p‐selectin can be used to detect platelet activation during plateletpheresis.
Efficacy of donor screening for HTLV‐I and the natural history of transfusion‐transmitted infection
Authors: Inaba S. , Okochi K. , Sato H. , Fukada K. , Kinukawa N. , Nakata H. , Kinjyo K. , Fujii F. , Maeda Y.
Published: March 2003

BACKGROUND: It has been 10 years since the implementation in Japan of donor blood screening for human T‐cell lymphotropic virus type I (HTLV‐I). This report reviews the effectiveness of screening in preventing transmission of HTLV‐I through blood transfusion and the current status of patients with confirmed seroconversion due to transfusions given before the implementation of screening.

STUDY DESIGN AND METHODS: Patients who received blood at Kyushu University Hospital from 1990 to 1997 were followed. Serum samples were collected before transfusion and 60 days or more after transfusion. Seroconversion was determined by a second‐generation particle agglutination test. Confirmation tests were an immunofluorescence assay, enzyme‐linked immunosorbent assay, and immunoblotting. Confirmed seroconverted patients were followed by a search of hospital records.

RESULTS: Seroconversion was found in one of 4672 transfused patients, but the donor was identified and confirmed to be negative for anti‐HTLV‐I and virus genome by nested polymerase chain reaction. A total of 23,323 red cell concentrates and 17,237 platelet concentrates were transfused to these 4672 patients. Therefore, the anti‐HTLV‐I prevalence in blood for transfusion after screening was estimated at 1 in 45,560 (0.0022%; the upper 95% CI was 0.0080%). One hundred two seroconverted patients who were transfused before donor screening for HTLV‐I were followed. One patient developed HTLV‐I‐associated myelopathy, diagnosed 18 weeks after seroconversion, and another patient developed uveitis 1 month after seroconversion. No patients developed adult T‐cell lymphoma, and the survival rate of seroconverted patients was 92.5 percent 15 years after transfusion.

CONCLUSION: This study confirmed that the present donor screening program for HTLV‐I by the new particle agglutination test can almost completely prevent virus transmission by transfusion. Complications of HTLV‐I transmission were at lower rates than expected.