bluuebunny/crossref_metadata_2025 huggingface dataset:
HLA antibodies as a cause of false‐positive reactions in screening enzyme immunoassays for antibodies to human T‐lymphotropic virus Type III
Authors: SAYERS MERLIN H. , BEATTY PATRICK G. , HANSEN JOHN A.
Published: March 2003

Abstract: 15,680 volunteer blood donors were screened by enzyme‐linked immunosorbant assay (EIA) for antibodies to human T‐lymphotropic virus Type III (HTLV‐III). On at least two of three determinations, 0.37 percent were found to be reactive. When the first 38 of these samples were sent for repeat EIA and Western blot, the EIA was positive in 14. In only three of these 14 samples was the EIA result confirmed by Western blot. Eight of these eleven false‐positive samples were from women who were found to have HLA antibodies. In seven of these women, the HLA antibodies were directed to Class II antigens expressed on the cell used to grow HTLV‐III in the preparation of screening EIA kits. We conclude that HLA antibodies are an important cause of false‐positive reactions in the screening test for HTLV‐III antibody.
In vitro function of granulocyte concentrates following passage through an electromechanical infusion pump
Authors: Snyder E. L. , Malech H. L. , Ferri P. M. , Gardner J. P. , Kalish R.
Published: March 2003

Six units of granulocytes stored for 18 hours at 20 to 24°C without agitation were passed through an electromechanical infusion device pump system (test) (Abbott) and through a 170‐μm filter (control). To determine the effect of needle gauge, a 19 or 23 gauge needle was attached to the end of the pump administration set. After passage through the pump, granulocytes were evaluated for signs of cytolysis and functional loss, red cells for hemolysis, platelets for release of β‐thromboglobulin, and plasma for C3a and C5a complement activation. There was no evidence (mean ± 1 SD) of hemolysis (<1.0 vs. <1.0 mg/dl), loss of granulocytes (123±38 × 103/μl vs. 118±29 × 103/μl), changes in any of several tests of neutrophil function, increased release of β‐thromboglobulin (15.8±5.8 vs. 17.0±6.6%), or C3a complement activation (483±221 vs. 500±200 ng/ml) after passage through the pump system (p>0.05). No significant differences were seen between the use of a 19 or 23 gauge needle. Based on these in vitro data, we conclude that this pump system is acceptable for use in clinical practice when control over rate and volume of granulocyte administration is important.
A simple method for preparing neocyte‐enriched leukocyte‐ poor blood for transfusion‐dependent patients
Authors: Hogan V. A. , Blanchette V. S. , Rock G.
Published: March 2003

Recent treatment for patients with thalassemia and chronic anemia involves transfusion of young red cells (YRBCs) or “neocytes.” We developed a technique enabling YRBCs to be separated based on their buoyant density in autologous plasma during centrifugation. Following this procedure, measurement of pyruvate kinase (PK), an age‐dependent red cell enzyme, showed neocyte enrichment in the top one‐third of the RBC layer corresponding to a mean of 47.5 percent of the total PK present in the unfractionated unit. To provide a neocyte transfusion preparation with an acceptable hemoglobin content, the top one‐third fraction from each of three bags of blood was pooled. Leukocytes were removed from this “neocyte unit” by an initial sedimentation with 6 percent hydroxyethyl starch (HES) followed by filtration through a filter (IG 500, Imugard). This process resulted in removal of 99.3 ± 0.5 percent (mean ± SD) of the leukocytes with a mean RBC recovery of 89.5 ± 5.5 percent and a final hemoglobin content of 53.4 ± 2.3 g. Tests for plasma‐free hemoglobin and HES in the supernatant of the final transfusion product gave acceptable mean values of 28 mg per dl and 3.0 mg per ml. Autologous mean RBC survival of Cr51 ‐labeled YRBC fractions was 41.8 ± 2.9 days (n=5).

This technique yields neocyte enrichment superior to that achieved using a cell processor (model 2991, IBM) and has the added advantage of being less costly to prepare ($45.00 [1984] U.S. per YRBC unit as compared to an estimated $135.00 [1984] U.S., IBM) and more economical in terms of blood use.
A multicenter trial to document the efficacy and safety of a rapidly excreted analog of hydroxyethyl starch for leukapheresis with a note on steroid stimulation of granulocyte donors
Authors: Strauss R. G. , Hester J. P. , Vogler W. R. , Higby D. J. , Snikeris A. C. , Imig K. M. , Greazel C. , Mallard G. , Burnett D. , Gupta S. , Hulse J. D.
Published: March 2003

Currently, the frequency of granulocyte donation is limited by the prolonged circulation of hydroxyethyl starch (HES). We conducted a Phase I, uncontrolled, multicenter trial to evaluate both the efficacy and safety of a rapidly excreted low‐molecular‐weight analog of HES (LMW‐HES). Seventy‐five donors underwent 179 centrifugation leukapheresis procedures using LMW‐HES as the red‐cell‐sedimenting agent. The efficacy of LMW‐HES was established by harvesting adequate numbers of leukocytes. Most granulocyte concentrates contained at least 20 × 109 neutrophils when 8 I of blood was processed from donors optimally stimulated with steroids. The safety of LMW‐HES was documented by the detection of almost no clinically significant adverse effects. In only 1.7 percent of procedures did donors require special attention, and only 1 of 179 procedures (0.6%) was permanently discontinued. Results of laboratory studies were usually normal or their values decreased transiently (approximately 15–25%) as a consequence of plasma volume expansion (dilution). Based on previous experience with HES, LMW‐HES and HES perform comparably during leukapheresis. When commercially available, use of this new, rapidly excreted analog should permit more frequent leukapheresis donation.
Liquid storage of unseparated human bone marrow. Evaluation of hematopoietic progenitors by clonal assay
Authors: Lasky L. C. , McCullough J. , Zanjani E. D.
Published: March 2003

Marrow from five donors was aspirated into a small volume of heparinized culture media, filtered, and stored in 10‐ml nominal plastic bags at 4, 22, and 37°C, respectively, for 7 days with and without agitation. Samples were taken at 0, 1, 2, 3, and 7 days, respectively, for determination of cell numbers, trypan blue viability, and committed colony‐forming unit‐granulocyte‐monocyte (CFU‐GM) and erythroid burst‐forming unit (BFU‐E) and colony‐forming unit (CFU‐E), as well as multipotent (CFU‐MIX), hematopoietic progenitors. Glucose and pH decreased with storage, most markedly at higher storage temperatures. Recovery of recovered cells exceeded 90 percent under all conditions at 7 days. Viability of committed and multipotent progenitors was maintained fairly well for 3 days at 4 and 22°C, but fell off rapidly after 1 day at 37°C. Recovery of progenitor cells stored 7 days was relatively poor. Best maintenance was achieved for 3 days using 4°C storage without agitation.
Inactivation of human T‐cell lymphotropic virus, type III by heat, chemicals, and irradiation
Authors: QUINNAN GERALD V. , WELLS MARTHA A. , WITTEK ALEC E. , PHELAN MICHAEL A. , MAYNER RONALD E. , FEINSTONE STEPHEN , PURCELL ROBERT H. , EPSTEIN JAY S.
Published: March 2003

Abstract: Infectivity of human T‐cell lymphotropic virus, Type III (HTLV‐III) was inactivated by heat more rapidly if in liquid medium than if lyophilized and more rapidly at 60° than 56°C. When HTLV‐III was added to factor VIII suspension, then lyophilized and heated at 60°C for 2 hours or longer there was elimination of 1 × 106 in vitro infectious units (IVIU) of virus. Much of the viral inactivation appeared to result from lyophilization. The application of water‐saturated chloroform to the lyophilized material containing virus also resulted in elimination of infectivity. HTLV‐III was efficiently inactivated by formalin, beta‐propiolactone, ethyl ether, detergent, and ultraviolet light plus psoralen. The results are reassuring regarding the potential safety of various biological products.
Look‐back: preliminary experience of AABB members. American Association of Blood Banks
Authors: BOVE J. R. , RIGNEY P. R. , KEHOE P. M. , CAMPBELL J.
Published: March 2003

By mid‐fall of 1986 the “look‐back” process conducted by blood banks belonging to the American Association of Blood Banks was about 50 percent complete. Over one‐half of the traced recipients had died; 15 percent were alive and negative for antibodies to the human immunodeficiency virus and 16 percent alive but had seroconverted. In a small number of cases physicians or patients elected not to be tested. No major problems were reported.
Intravenous immune globulin for patients alloimmunized to random donor platelet transfusion
Authors: Lee E.J. , Norris D. , Schiffer C. A.
Published: March 2003

Immune globulin, prepared as a chemically and enzymatically unmodified solution in 10 percent maltose at pH 4.25, was administered intravenously, at a dose of 0.4 g per kg per day for five consecutive days, to seven alloimmunized patients who had acute nonlymphocytic leukemia. All patients had an approximately threefold rise in IgG level. Five patients showed no change in lymphocytotoxic antibody (LCTAb) activity and no response to random donor platelets that were administered after the immune globulin. The activity of LCTAb either disappeared or diminished after immune globulin infusion in two patients, and they had borderline acceptable corrected count increments 1 hour after transfusion of pooled random‐donor platelet transfusions. Because of the diminished LCTAb activity, the improvements in response to pooled random‐donor platelet transfusions cannot be ascribed to the administration of immune globulin. High‐dose intravenous immune globulin has not been shown to be effective in reversing the effects of alloimmunization.
White cell fragmentation after therapeutic leukapheresis for acute leukemia
Authors: Van Rybroek J.J. , Olson J.D. , Burns C.P.
Published: March 2003

A 70‐year‐old woman with newly diagnosed acute nonlymphocytic leukemia (FAB M5) underwent therapeutic leukapheresis because of a white cell count (WBC) of 144 × 10(9) per I and clinical evidence of leukostasis. A peripheral blood film taken immediately after leukapheresis showed numerous cytoplasmic and nuclear fragments. The patient's clinical course thereafter was significantly compromised by disseminated intravascular coagulation with a severe bleeding diathesis, renal failure, and respiratory failure that led to her death. This case illustrates that therapeutic leukapheresis for elevated WBC in patients with acute leukemia may result in leukocyte fragmentation and possible intravascular coagulation.
Infection with hepatitis delta and human immunodeficiency viruses among hemophiliacs in Japan
Authors: Inagaki M. , Miura T. , Taki M. , Saito N. , Yamada K. , Unuma T. , Tsuda F. , Miyakawa Y. , Mayumi M.
Published: March 2003

Two hundred two patients with hemophilia, dependent solely on imported coagulation factor concentrates, were tested for markers of hepatitis B virus infection, antibody to hepatitis delta virus (anti‐HD), and antibody to human immunodeficiency virus (anti‐HIV). Nine carriers of hepatitis B surface antigen (HBsAg) were identified. Six (66.7%) of them were positive for anti‐HD, a prevalence much higher than that in HBsAg carriers without hemophilia in Japan (1/113 or 0.9%, p < 0.001). Anti‐HIV was found in 96 (47.5%), in sharp contrast to the low prevalence (0/1205) in apparently healthy blood donors (p < 0.001). These results implicated imported plasma products in the transmission of both delta and human immunodeficiency viruses to hemophiliacs. An efficient method for the sterilization of plasma products is warranted to prevent exposure of hemophiliacs to the accompanying pathogenic viruses.
Standardization of papain reagents by measurement of active sites using a synthetic inhibitor, E‐64
Authors: Scott M.L. , Whitton C.M.
Published: March 2003

L‐trans‐epoxysuccinyl‐leucylamido (4‐guanidino) butane (E‐64) reacts rapidly and irreversibly in a one‐to‐one ratio with the active site of papain, causing complete inhibition of the enzyme. After the addition of various concentrations of E‐64 to a papain preparation, the residual enzyme activity can be measured using an azoprotein technique. The molarity of E‐64 required to cause complete inhibition of papain activity is equal to the molarity of papain‐active sites. Preparations of papain from various sources were assayed for protease activity by hydrolysis of azoalbumin using several variants of the basic technique and also by hydrolysis of azocasein. For each variant of azoprotein assay procedure, the active sites of the papain were measured using E‐ 64. All variations of the azoprotein technique yielded similar estimates of the active site molarity of the papain preparations, whereas the azoprotein assay results alone showed wide variation. Quantitation of the active‐site molarity of various papain preparations using E‐64 correlated with serologic efficacy.
A chloroquine elution technique for platelet serology
Authors: Masel D.S. , Blumberg N. , Heal J.M.
Published: March 2003

The authors describe a prototype elution method employing chloroquine, a quinoline derivative, to elute IgG antibodies from the platelet surface. This chloroquine elution technique is relatively easy to perform and is effective in the removal of alloantibodies from the platelet surface. Eluted alloantibody was immunologically active once the chloroquine was removed from the eluate. The major advantage of this technique is that serologic testing of platelets after elution is possible, as 50 percent of the platelets remain after exposure to hypertonic acid chloroquine solution. Antigens on the platelet surface maintained their antigenicity subsequent to treatment, although measurable reductions in PIA1 reactivity occur. The elution technique was also successful in removing IgG from the platelet surface in patients with diseases involving elevated levels of platelet‐associated IgG.
Donor‐specific transfusions
Authors: Radvany R.M. , Patel K.M.
Published: March 2003

HLA profiles of 25 donor‐specific transfusion (DST) kidney donor‐ recipient pairs were analyzed for HLA antigen compatibility. Serum samples collected during and after DST were tested for cytotoxic antibodies against T and B lymphocytes of the donors and 30 normal individuals. Eleven recipients did not produce cytotoxic antibodies to the antigens of their DST donors, and eight produced cold and/or warm, broadly reactive B‐cell antibodies. Six patients (24%) produced HLA‐A, B, C, and/or DR antibodies. Three of these individuals produced antibodies after two immunizations, while others required three immunizations. Three of the 11 antibody nonproducers (17%) had not received previous transfusions, as compared to three of the eight antibody producers (43%). Comparison of HLA profiles revealed 22 percent of the HLA‐A, B, DR identities between the transfusion donor and recipient in antibody nonproducers as compared to 9 percent of the HLA‐A, B, DR identities in antibody producers. The HLA‐A2, B40, DR4 haplotype and HLA‐DRW6 antigen were more common among antibody producers than among nonproducers, who had an excess of the HLA‐B8, DR3 haplotype. These results are consistent with the hypothesis that there may be high‐ and low‐responder HLA haplotypes that control immunologic responsiveness to histocompatibility antigens.
The in vitro evaluation of two filters (Erypur and Imugard IG 500) for white cell‐poor platelet concentrates
Authors: Brubaker D.B. , Romine C.M.
Published: March 2003

White cell‐poor blood components are useful in patients with white cell antibodies. White cells are efficiently removed by two different filters, Imugard and Erypur, which have used saline as the filter solution. This study evaluated these filters as to their production of white cell‐poor platelets. Pools of random‐donor platelet concentrates were filtered. Prefiltration and postfiltration samples were evaluated for percentages of platelet recovery, white cell (WBC) removal, and platelet function. The two filter solutions tested were normal‐strength saline (NSS) and fresh‐frozen plasma (FFP). Postfiltration samples using NSS showed no measurable platelet aggregation with ADP, epinephrine, or collagen. However, with FFP, both filters showed 100 percent platelet aggregation with ADP, epinephrine, and collagen. The FFP filter solution provided excellent white cell removal in both filters (Imugard: 100% WBC removal or < 1.0 × 106 residual WBC; Erypur: 99.5% removal or > 1.0 × 107 residual WBC); however, platelet recovery was better with Imugard (95%) than with Erypur (55%). The filtration procedure is an excellent method for the preparation of white cell‐poor platelets; however, the quantity of the saline solution recommended for the filtering of red cells must be minimized for platelets.
Red cell alloimmunization in multitransfused HLA‐typed patients
Authors: Brantley S. G. , Ramsey G.
Published: March 2003

The authors studied retrospectively the formation of clinically significant red cell (RBC) alloantibodies in 958 HLA‐typed, multiply transfused patients receiving kidney (603 patients) or liver (263 patients) transplants or plateletpheresis transfusions (92 patients). RBC alloantibodies were found in 91 (9.5%) of these patients and multiple antibodies in 35 (3.7%). Rh (D, C, c, and E) antibodies accounted for 49 percent of the total and Kell antibodies for 31 percent. Antibodies were found in 15 percent of apheresis recipients and in 8.6 percent of renal and 9.5 percent of liver transplantation patients. No association was found between any HLA‐A, HLA‐B, or HLA‐DR phenotype and the presence of RBC alloantibodies, either in general or when analyzed according to the specific antibody. Renal transplant patients with RBC alloantibodies were somewhat more broadly HLA‐ alloimmunized than were those without RBC alloantibodies. Patient gender did not affect these results. The authors concluded that the immune response to RBC alloantigens is independent of HLA type but is associated with an increased level of HLA antibody formation.
Apheresis experience with a continuous flow cell separator
Authors: Norol F. , Aubert C. , Scotto F. , Duedari N.
Published: March 2003

The Dideco Vivacell separator is a continuous‐flow centrifugation system that has only recently been used for cytapheresis. The authors’ experience with this separator in 451 plateletpheresis and 164 leukapheresis procedures is presented. Platelet collection provided high platelet yields (9.53 ± 2.85 × 1011) with a collection efficiency of 74 ± 14 percent for about 6 liters of total blood processed. Functional integrity was confirmed by normal in vitro tests (aggregation and response to hypotonic stress) and good in vivo recovery (55%). In leukapheresis, white cell yields were high (3.42 ± 1.2 × 1010) with 85 percent polymorphonuclear neutrophil cells. Their oxidative metabolism functions (generation of free oxygen radicals), investigated by chemiluminescence, were increased over donor values. Donor reactions, all of the mild citrate type, were rare.
Quantitative studies on the D antigen of red cells with the Du phenotype
Authors: Szymanski I. O. , Araszkiewicz P.
Published: March 2003

Rh‐positive red cells (RBCs) that fail to agglutinate with commercial anti‐D reagents by the tube method are considered to have the Du phenotype. The quantities of D epitopes on such RBCs have been measured previously in one kindred. The authors report on the number of D epitopes on Du RBCs of 23 unrelated individuals, as calculated by Scatchard's analysis. Cell‐bound anti‐D was measured by an automated antiglobulin consumption technique. On the average, RBCs of the Rh phenotype CcDue had a mean of 1568 ± 1220 (n = 12) D epitopes per cell. The relatively large range of values in this group implies a heterogeneous genetic background. The lowest number of D epitopes, 285 per cell, was observed on the RBCs of one individual who was apparently homozygous for C. In this case, the D antigen was detected only by adsorption/elution tests. RBCs of the phenotype cDuEe had a mean of 775 ± 378 epitopes per cell (n = 8), and those from two individuals with phenotype cDue had 2840 and 1560 D epitopes, respectively. Thus, on the average, RBCs with the Du phenotype bear about 10 to 20 times less D antigen than normal Rh‐positive RBCs. It is suggested that the low D antigen density of Du red cells may account for their poor immunogenicity.
The pharmacology of a new pasteurized antihemophilic factor concentrate derived from human blood plasma
Authors: Rousell R. H. , Kasper C. K. , Schwartz R. S.
Published: March 2003

The pharmacology of a new pasteurized factor VIII (FVIII) concentrate derived from human blood plasma was studied in 23 adults with hemophilia A. In Part 1 of the study involving six nonbleeding subjects, the mean increase in FVIII activity was 1.43 ± 0.34 U per ml 10 minutes after an intravenous dose of 50 U per kg. The intravascular survival kinetics in these six patients showed a biphasic decay curve with an initial mean half‐life of 5.1 ± 1.2 hours probably representing early redistribution, and a late half‐life of 13.3 ± 4.9 hours. In Part 2 of the study, the activity at 10 minutes was measured in another 17 patients, as well as in one patient already studied in Part 1. The mean increase in activity with the 24 observations was 1.13 ± 0.37 U per ml with a mean FVIII dosage of 51.0 ± 2.6 U per kg of body weight. Only one patient had an allergic reaction, which did not recur when the patient was given a second lot.
The effect of additives on red cell 2,3 diphosphoglycerate levels in CPDA preservatives
Authors: Vora S. , West C. , Beutler E.
Published: March 2003

Forty‐two chemical substances, chosen because they might influence red cell metabolism, were screened for effect on red cell adenosine triphosphate and 2,3 diphosphoglycerate (2,3 DPG) levels during storage in CPD or CPDA‐1 at 4° C. Of these substances, six appeared on initial screening to increase 2,3 DPG levels during storage; on repeated examination, four compounds, i.e., oxalate, glyoxalate, ethyl oxaloacetate, and L‐phenylalanyl‐L‐alanine, consistently increased 2,3 DPG levels during storage. It was shown that glyoxalate was converted rapidly to oxalate in blood, presumably through the lactate dehydrogenase reaction. Ethyl oxaloacetate is known to hydrolyze, giving rise to oxalate. Thus, the effect of both glyoxalate and ethyl oxaloacetate can be explained by the formation of oxalate, a compound already known to increase 2,3 DPG levels. The effect of L‐phenylalanyl‐L‐alanine remains to be explained, but it may be hydrolyzed to L‐alanine and L‐phenylalanine, both of which are thought to have the capacity to increase red cell 2,3 DPG levels by inhibiting pyruvate kinase activity.
American Red Cross experience with routine testing for hepatitis B core antibody
Authors: AuBuchon J. P. , Sandler S. G. , Fang C. T. , Dodd R. Y.
Published: March 2003

The implementation of routine testing of blood donations for hepatitis B core antibody (anti‐HBc) has allowed the characterization of the performance of the test in a large number of samples from apparently healthy individuals. This study reports the experience of the American Red Cross in testing 2.3 million donors for anti‐HBc. The test protocol reproducibly identified a distinct population of donors. The anti‐HBc‐positive rate varied by region of the continental United States and by the time of year. In a case‐control study, 85 percent of subsequent donations from anti‐HBc‐positive donors were anti‐HBc positive. The predictions made in an earlier pilot study regarding the performance and impact of the test were borne out.
Attitudes and physical condition of unrelated bone marrow donors immediately after donation
Authors: Stroncek D. , Strand R. , Scott E. , Kamstra‐halvorson L. , Halagan N. , Rogers G. , Mccullough J.
Published: March 2003

Annually, over 3000 bone marrow transplants are performed worldwide involving HLA‐identical sibling donors. However, only 30 percent of those patients who need a bone marrow transplant have a matched sibling donor. Programs have been developed to provide volunteer unrelated bone marrow donors for patients without sibling donors. Because bone marrow donation requires a high level of altruism, especially on the part of a donor unrelated to the patient, it is important to determine the effect of donation on the donor. Serious medical complications during the collection of marrow from related donors have been reported only rarely, but there have been no reports on the psychosocial and physical effects of the bone marrow donation process on volunteer unrelated bone marrow donors. The first 20 unrelated volunteers who donated bone marrow through the authors' volunteer bone marrow donor program were surveyed. They suffered no serious emotional or physical aftereffects. Seventeen of 20 donors reported that making the decision to donate was easy, and 16 reported making the decision to donate right away. Nine of 20 donors, however, stated that a friend or family member discouraged them from donating. All 20 donors thought that they were well prepared for the donation process. Four donors thought that the donation process required more time than they expected, but four donors thought the reverse. One person thought the process was more painful than they expected; however, 12 found the process to be less painful than expected. Nineteen of the 20 donors said that they would donate bone marrow again, and 17 would advise others to donate marrow. No life‐threatening medical complication occurred; however, one donor (5%) experienced orthostatic hypotension for 1 day following the collection. No donors received homologous blood transfusions, and all donors received 1 or 2 units of autologous red cells. No contraindication to the continued use of unrelated volunteer bone marrow donors was found.
Applications of ultraviolet light in the preparation of platelet concentrates
Authors: Pamphilon D. H. , Corbin S. A. , Saunders J. , Tandy N. P.
Published: March 2003

Passenger lymphocytes in platelet concentrates (PCs) may induce the formation of lymphocytotoxic antibodies (LCTAbs) and subsequent refractoriness to platelet transfusions. Ultraviolet (UV) irradiation can prevent lymphocytes' acting as stimulator or responder cells in mixed‐lymphocyte reactions (MLRs) and could theoretically prevent LCTAb formation in vivo. A system has been devised for the delivery of UV irradiation to PCs: platelet storage characteristics and MLRs were evaluated in UV‐irradiated PCs harvested from healthy donors with the Haemonetics V50 and PCS cell separators. MLR and response to phytohemagglutinin stimulation were abolished by a dose of 3000 joules per m2 at a mean wavelength of 310 nm. Platelet aggregatory responses to adenosine diphosphate (ADP), ristocetin, collagen and epinephrine, hypotonic shock response, and pH showed no important differences when control PCs and PCs irradiated as above were compared during 5 days of storage in Fenwal PL‐1240 packs. Lactate production during storage was significantly higher in UV‐treated PCs (p<0.001), but values did not exceed 20 mmol per L. UV transmission at 310 nm in standard blood product containers, including the Fenwal PL‐146, PL‐1240, and PL‐732, was low (<30%), but it was acceptable in the Delmed Cryostorage and DuPont SteriCell packs (>50%). UV irradiation may provide a simple and inexpensive means of producing nonimmunogenic PCs.
Plasma levels of ionized and total calcium during storage of citrated platelet concentrate
Authors: Bode A. P. , Miller D. T. , Toffaletti J.
Published: March 2003

Measurements of ionized and total calcium levels in supernatant plasma samples from citrated platelet concentrates (PCs) were made over 7 days of storage. Both ionized and total calcium increased significantly during the storage period: respectively, from 0.074 mM Ca2+ in fresh platelet‐rich plasma to 0.084 mM in PCs stored for 7 days (p = 0.017), and from 1.94 mM total calcium to 2.06 mM (p = 0.014) over the same period. The increase in calcium was partially blocked by the addition of platelet activation inhibitors to the PCs. Platelet‐poor plasma stored under similar conditions showed no significant change in ionized or total calcium, which indicated that the increases observed in PCs were due to the release of cellular calcium. Significant correlations (p<0.01) were found between ionized or total calcium levels and lactate concentration or pH, but not hypotonic shock recovery rate. The demonstration of non‐zero levels of ionized calcium makes it likely that Ca2+‐dependent enzyme systems such as calpain expression and thrombin generation are active in the plasma of citrated PCs and may contribute to the platelet storage lesion.
Differences in the susceptibility of platelets to freezing damage in relation to size
Authors: van Prooijen H. C. , van Heugten J. G. , Riemens M. I. , Akkerman J. W. N.
Published: March 2003

Previous studies have shown that cryopreservation of normal platelets induces a reduction in the contents of secretion granules, the generation of thromboxane B2, and aggregability. The present study investigates whether these changes in the total population occur to the same extent in four size‐dependent subpopulations with mean platelet volumes of 4.2, 5.7, 8.0, and 11.1 μm3, obtained by counterflow centrifugation. Cryopreservation reduced the contents of the alpha granule markers and the generation of thromboxane B2 in the platelets from the four fractions to the same extent as in the platelets from the total suspension. Maximal aggregation of the platelets in response to collagen was measured by optical aggregation. The average decrease in light transmission after freezing was 47 ± 3 percent (SEM) for the platelets in the total population, 40 ± 3 percent for the largest platelets, and 65 ± 5 percent for the smallest platelets, which indicates that aggregability was better preserved in the larger platelets than in the smaller cells. It is possible that, in the smallest platelets, a decrease in thromboxane generation of approximately 70 percent becomes rate‐limiting for aggregation. Further improvements in the clinical use of freeze‐preserved platelets may be sought in the preparation of concentrates with relatively high counts of large platelets.
Growth of gram‐positive and gram‐negative bacteria in platelet concentrates
Authors: Punsalang A. , HealMBBS J. M. , Murphy P. J.
Published: March 2003

In 1986 the allowable platelet storage time was reduced from 7 to 5 days because of a recent increase in septic deaths associated with platelet transfusion. In this study, the growth curves of two gram‐positive and two gram‐negative organisms in platelets stored for 7 days in CLX and PL‐732 bags were evaluated. Platelets in CLX bags were inoculated with 101, 102, and 103 organisms and 102 organisms were introduced into PL‐732 bags. Test organisms were inoculated into trypticase soy broth as a control. All four bacteria grew rapidly in trypticase soy broth, reaching 109 organisms per mL within 48 hours. In both CLX and PL‐732 bags, the growth pattern of gram‐positive organisms was generally logarithmic during the first few days of storage. A concentration of 108 organisms per mL was present by Day 3 or 4, after which further proliferation was inhibited by the high density of bacteria in the platelets. In PL‐732 bags, the proliferation of gram‐negative organisms followed a pattern similar to that of the gram‐positive bacteria. However, gram‐negative organisms grew less well in CLX bags. Their growth was either delayed, inhibited after a few days, or entirely suppressed. Our data suggest that 1) changing the storage period of platelets from 7 to 5 days is unlikely to reduce the incidence of clinical sepsis caused by gram‐positive organisms if the number of organisms introduced at the time of contamination is on the order of 10 to 1000 bacteria; 2) gram‐negative organisms tend to proliferate more slowly in CLX bags than in PL‐732 bags; and 3) shortening the shelf life of platelets to 5 days may not eliminate gram‐negative sepsis after transfusion of platelets stored in PL‐732 bags, but it may reduce the number of such episodes associated with CLX bags.
The Kmod blood group phenotype in a healthy individual
Authors: Winkler M. M. , Beattie K. M. , Cisco S. L. , Sigmund K. E. , Johnson C. L. , Rabin B. I. , Marsh W. L.
Published: March 2003

This report describes a healthy blood donor whose red cells have weakened expression of Kell blood group antigens. Kell antigen activity could not be detected by flow cytometric analysis and was demonstrable only by sensitive serologic techniques. As with normal‐strength Kell antigens, reactivity could be abolished by treatment with 2‐aminoethylisothiouronium bromide (AET). The donor's red cells have Kx antigen activity. Other commonly tested blood group antigens (MNSs, Rh, P1, Lewis, Duffy, and Kidd systems) appear normal. Clinical and serologic examination showed that this case is different from previously described examples of modified Kell expression. The propositus's phenotype has remained unchanged for 19 months, which suggests that it is not a transient condition. However, family studies provide no evidence that it is inherited. A 93‐kD protein, which reacted weakly by Western blot with rabbit antibody to Kell protein, was isolated from the propositus's red cells by immunoprecipitation. This finding was not reproduced in subsequent studies, which suggests that the quantity of Kell protein recovered was at the threshold level detectable by the technique used. The red cell phenotype is categorized as Kmod, of which this is the first example reported in a healthy individual.
Filtration. A method to prepare white cell‐poor platelet concentrates with optimal preservation of platelet viability
Authors: van Kooy Marinus Marwijk , van Prooijen Hendrik C. , Borghuis L. , Moes Marcel , Akkerman Jan‐Willem N.
Published: March 2003

HLA alloimmunization is a major problem for thrombocytopenic patients receiving long‐term platelet support. It is caused by white cells (WBCs) that are present as contaminants in platelet concentrates (PCs). Recent data have shown that filtration is an effective means to reduce WBC contamination, but it has little effect on the recovery of platelets. The present report evaluates two filters, a cellulose acetate (CA) filter requiring the inactivation of platelets with prostacyclin and a cotton wool (CW) filter requiring no platelet inactivation. The results show that, using fresh pooled PCs from six random donors, both filters reduce WBC contamination below 10(7) per PC, the likely threshold below which alloimmunization does not develop. With platelets stored for 2 to 3 days the efficacy of the CW filter decreases. Neither filter inflicts important damage to the platelets, as there is no considerable platelet activation or cell disruption. Moreover, PCs prepared by both filters show normal survival and effectively reduce the bleeding times. Thus, filtration of PCs results in platelets with optimal responsiveness both in vitro and in vivo.
When should antibody screening tests be done for recently transfused patients?
Authors: Shulman Ira A. , Nelson Janice M. , Nakayama Roland
Published: March 2003

The American Association of Blood Banks (AABB) requires that blood samples used for pretransfusion testing of recently transfused (or pregnant) patients must be obtained within 3 days of scheduled transfusions. This requirement, which became effective in July 1988, amended Standard G2.000 of the AABB, which previously required that pretransfusion testing must be done on blood samples obtained within 2 days of scheduled transfusions. The present study was designed to estimate the risk associated with adopting the amended version of Standard G2.000. Sixty patients who developed significant unexpected alloantibodies after transfusion were studied retrospectively. Thirteen of the 60 patients were found to have newly detectable antibodies within 83 hours of a sample reported to be negative for the new antibody. Had the amended version of Standard G2.000 been in effect, the detection of some of these antibodies might have been delayed up to 24 hours. It was estimated that the implementation of the new AABB requirement at the authors' institution could potentially place about 1 in 3000 transfused patients at risk for an acute or delayed hemolytic transfusion reaction.
Transfusion‐related acute lung injury caused by an NB2 granulocyte‐ specific antibody in a patient with thrombotic thrombocytopenic purpura
Authors: Van Buren Nancy L. , Stroncek David F. , Clay Mary E. , McCullough Jeffrey , Dalmasso Agustin P.
Published: March 2003

HLA and granulocyte‐specific antibodies have been implicated in the production of transfusion‐related acute lung injury (TRALI). Reported here is a case that suggests that the patient's preexisting condition may play an important role in determining whether TRALI develops upon transfusion of blood products containing anti‐white cell (WBC) antibodies. A 29‐year‐old woman with thrombotic thrombocytopenic purpura (TTP) underwent an uneventful 1.5‐volume plasma exchange, which was followed by the transfusion of 2 red cell (RBC) units. At the end of the second RBC transfusion, the patient developed clinical signs and symptoms of noncardiogenic pulmonary edema. Serologic studies demonstrated that the serum from the second RBC donor had no HLA antibodies but did have a granulocyte‐specific antibody (anti‐NB2) that caused the agglutination of the recipient's granulocytes, which were NB2 positive. Serum from the donor of the first RBC unit and serum from the donors of units used in the exchange had no HLA or granulocyte‐ specific antibodies that reacted with the recipient's WBCs. Because the donor implicated in this reaction had a history of 21 blood donations, none of which had been associated with a transfusion reaction, we suggest that the patient's preexisting condition played a significant role in this episode of TRALI, owing to the granulocyte‐specific antibody.
EFFECT OF BLOOD TRANSFUSIONS ON EXPERIMENTAL PULMONARY METASTASIS IN MICE
Authors: SHIRWADKAR S. , BLAJCHMAN M.A. , FRAME B. , ORR F.W. , SINGAL D.P.
Published: March 2003

We examined the effect of allogeneic blood transfusions (BT) on pulmonary metastases in a mouse model. Recipient (C57B1/6J) mice were transfused with either saline, syngeneic blood or allogeneic (Balb/c) blood on two occasions, days 0 and 3. One week after the last transfusion, recipient mice were injected intravenously with varying numbers of methylcholanthrene‐induced fibrosarcoma cells. Twenty days later the number of pleural nodules was counted as an index of pulmonary metastasis. The data demonstrate that the inoculation of 2.5 times 105 or 1 times 105 tumor cells resulted in significantly higher numbers of pulmonary metastases in mice that received allogeneic BT than the mice that received syngeneic blood or saline. In contrast, allogeneic BT caused significant inhibition of pulmonary metastases in mice that received 3.5 times 105 tumor cells. The data suggest that the immunomodulatory (stimulatory or inhibitory) effect of BT is dependent on the numbers of tumor cells inoculated. It is likely that the conflicting reports in the literature on the effects of BT on tumor growth may be due to inoculation of different numbers of tumor cells. These results have an important bearing in understanding the effect of allogeneic BT on tumor growth both in experimental animals and in cancer patients.
The in vivo survival of human red cells with low oxygen affinity prepared by the osmotic pulse method of inositol hexaphosphate incorporation
Authors: Franco R. , Barker R. , Mayfield G. , Silberstein E. , Weiner M.
Published: March 2003

The purpose of this study was to determine the posttransfusion survival of low‐affinity red cells (RBCs) containing the hemoglobin modifier, inositol hexaphosphate (IHP). An osmotic pulse technique was used to incorporate IHP, a compound that normally does not cross the RBC membrane. Two groups of six volunteers were studied. RBCs were treated under conditions that resulted in a relatively large increase in mean corpuscular volume (Δ MCV = 11.8 ± 4.0, 1 SD) in Group I, but a relatively small increase (Δ MCV = 5.5 ± 1.9) in Group II. The groups had a similar decrease in mean corpuscular hemoglobin (Δ MCH = −3.8 ± 0.9 for Group I and −3.1 ± 1.1 for Group II). The 24‐hour posttransfusion RBC survival, measured with 3 mL of 51Cr‐labeled autologous RBCs, correlated with the induced change in oxygen affinity, and larger shifts were associated with poorer survival. An acceptable 75‐percent 24‐hour posttransfusion survival was achieved for p50 increases up to approximately 15 torr. Cells that survived 24 hours appeared to have a normal lifespan. This study is the first demonstration of adequate survival for human RBCs with a potentially long‐lasting decrease in oxygen affinity.
Humoral immune response following extracorporeal immunoadsorption therapy of patients with hypercholesterolemia
Authors: Gordon B.R. , Sloan B.J. , Parker T.S. , Saal S.D. , Levine D.M. , Rubin A.L.
Published: March 2003

Low‐density lipoprotein apheresis (LDL‐apheresis) is an extracorporeal procedure that preferentially removes LDL cholesterol from the blood. One of the primary techniques for performing this procedure uses immunoadsorption columns containing monospecific polyclonal sheep antibodies to human LDL covalently coupled to a gel filtration medium. LDL‐apheresis has generally been well‐tolerated, with chills, fever, or flushing occurring rarely. The possibility of an immune reaction was investigated as a basis for these reactions observed in 12 of the 1312 procedures performed. Antibodies to sheep IgG developed in 12 of the 15 patients treated with LDL‐apheresis as a result of the shedding of small quantities of the sheep immunoglobulin from the columns. A column acid‐washing procedure minimized the quantity of shed antibody but did not prevent immunization of the patient. The clinical reactions were probably unrelated to shedding and immunization, as the reactions occurred even in patients who were not immunized to the sheep IgG. Immunization to ethylene oxide was not the cause, as determined by a radioallergosorbent test. The reactions were more likely related to the activation of complement, as indicated by the generation of C3a des Arg by the columns and an increase in C3a des Arg levels systemically.
Reports of 355 transfusion‐associated deaths: 1976 through 1985
Authors: Sazama K.
Published: March 2003

From 1976 through 1985, the United States Food and Drug Administration received reports of 355 fatalities associated with transfusion, 99 of which were excluded from further review because they were unrelated to transfusion or involved hepatitis or acquired immune deficiency syndrome. Of the remaining 256 reported deaths, 51 percent resulted from acute hemolysis following the transfusion of ABO‐incompatible products. These deaths were due primarily to managerial, not clerical, errors. Other causes of death (in order of frequency of report) included acute pulmonary injury (15%), bacterial contamination of product (10%), delayed hemolysis (10%), damaged product (3%), and graft‐ versus‐host disease (0.4%). Management systems for transfusion facilities should be created or revised to include the specific identification of personnel eligible to administer transfusions to provide written guidance and appropriate training (including recognition and management of errors), and to implement measures that target safe transfusion practices. Continued research into acute pulmonary injury, the immunologic hazards of transfusion, and the prevention of bacterial contamination of blood components is necessary.
Minimal‐exposure transfusion and the committed donor
Authors: Brecher M.E. , Taswell H.F. , Clare D.E. , Swenke P.K. , Pineda A.A. , Moore S.B.
Published: March 2003

Autologous blood (collected preoperatively or salvaged intraoperatively) is the safest blood available for transfusion, but its use is not always feasible. It may be possible to decrease a patient's exposure to homologous donors. Pediatric cardiac surgery patients frequently are unable to donate autologous blood preoperatively. Since 1984, attempts have been made to provide parental apheresis platelets and intraoperative blood salvage to such patients to decrease their donor exposure. Further decreases in donor exposure have been the object of a program of collecting from one committed donor all the blood a patient is anticipated to need. This article reviews the experience with 50 pediatric cardiac surgery patients on such a program, in whom the mean decrease in homologous‐donor exposure was 57 percent (range, 12–93%). Thirteen of these patients received only homologous blood products from one committed donor, for a mean decrease in homologous‐donor exposure of 80 percent (range, 50–93%). A comparison of 12 of these 13 recipients with a matched control group showed no significant difference in red cell transfusion practice but a significant difference in the number of homologous‐donor exposures per m2 of body surface area (BSA) (mean donor exposures/m2 of BSA: patients = 1.5, controls = 10.5). The use of one committed donor and autologous blood can provide a minimal‐exposure transfusion.
Prevalence of anti‐HCV in Norwegian blood donors with anti‐HBc or increased ALT levels
Authors: Hetland Geir , Skaug K. , Larsen J. , Mæland A. , Strømme J.H. , Størvold G.
Published: March 2003

Testing for hepatitis B core antibodies (anti‐HBc) was performed in 12,104 consecutive blood donors, 139 (1.15%) of whom were found to be positive. The first 6036 donors were also screened for ALT; 91 (1.51%) had repeatedly elevated values. ALT screening was of no help in detecting anti‐HBc‐positive donors. Those with anti‐HBc or repeatedly raised ALT levels were further tested for hepatitis C virus antibodies (anti‐HCV), and 3 (2.16%) and 1 (1.10%) anti‐HCV‐positive donors were detected, respectively. This prevalence of anti‐HCV (0.5%) is not significantly different from that found in 1000 unselected donors at our blood bank. Testing for ALT and anti‐HBc as surrogate markers for hepatitis C is therefore not recommended in Norwegian blood donors and should be replaced by anti‐HCV screening.
American blood donors seropositive for human T‐lymphotropic virus types I/II exhibit normal lymphocyte subsets
Authors: Prince Harry E.
Published: March 2003

Recent reports have demonstrated that some lymphocyte subsets are abnormal in Japanese blood donors who are seropositive for human T‐ lymphotropic virus type I (HTLV‐I). To determine if similar changes characterize American blood donors who are seropositive for HTLV‐I/II, lymphocyte subsets were measured in 42 HTLV‐seropositive and 42 HTLV‐ seronegative blood donors. The seronegative individuals were matched by age, race, and gender to the seropositive individuals. Peripheral blood mononuclear cells were treated with a panel of 12 monoclonal antibody pairs and then analyzed by two‐color flow cytometry. No significant differences were observed between the seropositive and seronegative groups with respect to the absolute number of circulating lymphocytes or the percentages of lymphocytes belonging to the subsets assessed. These subsets included B, T, CD4, and CD8 cells and subpopulations of CD4 and CD8 cells defined by the coexpression of markers that appear (CD25, HLA‐DR, CD38) or disappear (Leu 8, CD45RA) after activation. These findings indicate that HTLV‐seropositive persons in the American blood donor pool do not exhibit the lymphocyte subset alterations reported for HTLV‐I‐seropositive blood donors in Japan.