bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunogenicity and antigenicity of defined linear determinants of the MN sialoglycoprotein glycophorin A
Authors: Rearden A.
Published: March 2003

Many epitopes of the blood group MN sialoglycoprotein glycophorin A (GPA) are clinically important and have been implicated in alloimmunization and autoimmune hemolytic anemia. However, the factors that contribute to the immunogenicity and antigenicity of GPA epitopes are poorly understood. To study this question, linear determinants were defined to include all GPA exofacial regions that were free of linked carbohydrate (GPA 27–34, GPA 52–62, and GPA 65–70), and peptides corresponding to these amino acids were synthesized. All defined GPA determinants were shown to contain residues accessible to the immune system. Immunogenicity was measured by binding rabbit antisera, produced by using red cells (RBCs), RBC membrane, and purified GPA as immunogens, to peptide antigens in an enzyme‐linked immunosorbent assay. GPA 27–34 and GPA 52–62 were shown to be immunogenic, but GPA 65–70 was not. Comparison of the binding of antisera to peptide and purified GPA antigens showed that the defined determinants were less immunogenic than other, nondefined GPA epitopes. Antigenicity algorithms (hydrophilicity, surface probability, flexibility, and antigenic index) predicted that GPA 27–34 and, to a lesser extent, GPA 52–62 would be antigenic, while GPA 65–70 would be nonantigenic. Antigenicity, measured by the binding of rabbit antipeptide sera to RBCs, RBC membrane, and purified GPA, was demonstrated for the GPA 52–62 determinant alone. Results differed among molecular forms of GPA, which indicated that the specific conformation assumed by GPA is important in determining the immunogenicity and antigenicity of its epitopes. GPA immunogenicity and antigenicity are distinct phenomena that do not always predict one another, and, while accessibility to the immune system is required for immunogenicity and antigenicity, it alone is insufficient for either.
Informed consent for blood transfusion: a regional hospital survey
Authors: Eisenstaedt R.S. , Glanz K. , Smith D.G. , Derstine T.
Published: March 2003

Ninety‐two hospitals in a three‐state mid‐Atlantic region were surveyed to determine their policy toward obtaining written informed consent for transfusion and to examine the content of written consent documents and the process by which consent is obtained. Of 81 hospitals responding, 50 (62%) required written informed consent. Hospitals with fewer than 200 beds were more likely to require written informed consent. The attending physicians had responsibility for obtaining consent in 28 (57%) of 49 institutions, most often on the day or evening before surgery. Twenty‐seven of 48 forms mentioned complications: hepatitis in 80 percent, human immunodeficiency virus infection in 46 percent, nonhemolytic reactions in 32 percent, and hemolysis in 25 percent. Alternatives to allogeneic transfusion were mentioned infrequently; eight hospital forms listed autologous transfusion options and only two mentioned designated donation. The reading level required to comprehend 34 consent forms submitted was grade 14.6, which has been attained by only 23 percent of the adult United States population. Although the majority of respondent institutions require written informed consent, those forms, per se, do not document that the fundamental tenets of informed choice have been applied to the decision to transfuse blood.
Characterization of the neutrophil molecules identified by quinine‐ dependent antibodies from two patients
Authors: Stroncek D. F. , Herr G. P. , Maguire R. B. , Eiber G. , Clement L. T.
Published: March 2003

BACKGROUND: Two patients with episodic pancytopenia and renal failure associated with quinine (Qn) ingestion were previously found to have Qn‐ dependent antibodies that reacted with red cells, platelets, and neutrophils. The purpose of these studies was to characterize the neutrophil antigens recognized by Qn‐dependent antibodies from these two patients.

STUDY DESIGN AND METHODS: The neutrophil molecules recognized by the Qn‐dependent antibodies in the sera from the two patients were analyzed by immunoprecipitation using 125I‐labeled neutrophils. Neutrophils from 13 different donors were tested.

RESULTS: The Qn‐dependent antibodies from Patient 1 immunoprecipitated a 60‐kDa molecule on neutrophils from seven donors and an 85‐kDa molecule on neutrophils from three donors. The Qn‐dependent antibodies from Patient 2 reacted with a 32‐kDa molecule on neutrophils from 5 donors, a 60‐kDa molecule on neutrophils from 9 donors, and an 85‐kDa molecule on neutrophils from 10 donors. Neutrophil‐specific antigen NB1 is also located on a 60‐kDa glycoprotein (GP). While the antibody in serum from Patient 1 did not show specificity for NB1, the antibody from Patient 2 detected the 60‐kDa molecule on NB1‐positive neutrophils from 9 of 11 donors tested and did not detect the 60‐kDa molecule on NB1‐negative neutrophils from 2 donors. In a monoclonal antibody immobilization of granulocyte antigens assay, the Qn‐dependent antibody from both patients reacted with the 60‐kDa molecule carrying NB1. The Qn‐ dependent antibody from a third patient, Patient 3, was previously found to react with an 85‐kDa GP and the 60‐kDa NB1 GP. To determine if the Qn‐dependent antibodies from Patients 2 and 3 recognized the same 85‐kDa GP, neutrophils were treated with serum from Patient 3 plus Qn to remove the 85‐kDa GP. Then, serum from Patient 2 plus Qn no longer immunoprecipitated the 85‐kDa GP.

CONCLUSION: The antigens recognized by Qn‐dependent neutrophil antibodies were located on molecules of 85, 60, and 32 kDa. Qn‐dependent antibodies from two patients reacted with the same 85‐kDa GP and those from three patients reacted with the same 60‐kDa GP. The 60‐kDa molecule recognized by the Qn‐dependent antibodies carried the NB1 antigen.
Evaluation of platelet concentrates stored for 5 days with reduced plasma volume
Authors: Holme S. , Heaton W.A. , Moroff G.
Published: March 2003

BACKGROUND: Currently, platelet concentrates (PCs) are stored in a suspending plasma volume of 45 to 65 mL. Previous studies using second‐ generation containers indicated that PCs stored for 5 days at volumes less than 30 mL have reduced in vivo percentage recoveries as compared to PCs stored at volumes of 50 mL or more.

STUDY DESIGN AND METHODS: This study has evaluated the effect of PC plasma volume on the maintenance of in vivo and in vitro platelet properties following 5 days of storage, with the purpose of establishing the minimum plasma volume in the range of 30 to 50 mL. Twenty paired studies were performed in which identical populations of platelets from the same donor (obtained by double manual apheresis) were stored in a normal volume (55‐60 mL, control) and reduced volume (30‐50 mL, test) of plasma. Comparison of in vivo viability between test and control PCs was performed after random radiolabeling of 1 unit with 51Cr and of the other with 111In, with simultaneous transfusion and with calculation of percentage recovery and the area below the survival curve (integral) as measures of viability.

RESULTS: When test unit volumes were > or = 35 mL, essentially identical platelet survival curves and in vitro results were obtained for test and control. The integral and the percentage recovery for the test units were (mean, 95% confidence interval) 98.7 (96.3‐101.0) and 99.0 percent (94.7‐103.3) of those values in the control units, respectively. Test units with volume < or = 34 mL demonstrated reduced in vivo viability with integral and percentage recovery of 81.1 (68.9‐93.3) and 80.4 (69.3‐91.5), respectively, as compared to the control units. This loss was associated with increased metabolic activity (lactate production), which may suggest platelet activation due to the increased surface‐to‐PC volume ratio.

CONCLUSION: These results show that the storage volume of PCs may be reduced from 50 to 60 mL to 35 to 40 mL without any significant decrease in in vivo or in vitro platelet quality.
In vitro production of interleukin‐1 receptor antagonist in IgG‐ mediated red cell incompatibility
Authors: Davenport R.D. , Burdick M.D. , Strieter R.M. , Kunkel S.L.
Published: March 2003

Background: Recently, proinflammatory cytokines including interleukin 1 (IL‐1) have been implicated in the pathophysiology of immune‐mediated hemolysis. Little is known, however, about the mechanisms by which inflammatory events in these reactions may be downregulated. IL‐1 receptor antagonist (IL‐1ra) inhibits the actions of IL‐1 by competition for cellular receptors, and thus it may regulate the biologic actions of IL‐1 during immune‐mediated hemolysis. The production of IL‐1ra by peripheral blood mononuclear cells (PBMNCs) in response to IgG‐coated red cells in vitro was investigated.

Study Design and Methods: Fresh PBMNCs were obtained by density gradient separation of heparinized whole blood. PBMNCs were cultured in the presence of anti‐D‐coated, Rh‐positive red cells or uncoated Rh‐ negative red cells under nonadherent conditions. IL‐1ra concentrations in the culture media were measured by enzyme‐linked immunosorbent assay. IL‐1ra and IL‐1β gene expression was assessed by Northern blot analysis of PBMNC mRNA.

Results: IL‐1ra production was evident 4 hours after stimulation with IgG‐coated red cells and increased progressively over 24 hours. Gene expression of IL‐1ra was first detected at 2 hours, lagging behind that of IL‐1β. IL‐1ra gene expression was not inhibited by neutralizing polyclonal antibodies to IL‐1. Immunocytochemical staining to determine the cellular source localized IL‐1ra production to monocytes engaged in erythrophagocytosis. IL‐1ra production was inhibited by dexamethasone (10−7 M).

Conclusion: These results suggest that IL‐1ra production may partly account for the variable pathophysiologic events seen in IgG‐ mediated autoimmune hemolysis and hemolytic transfusion reactions. Steroid treatment may also downregulate anti‐inflammatory cytokine production in immune hemolysis.
Laboratory diagnosis of heparin‐associated thrombocytopenia and comparison of platelet aggregation test, heparin‐induced platelet activation test, and platelet factor 4/heparin enzyme‐linked immunosorbent assay
Authors: Greinacher A. , Amiral J. , Dummel V. , Vissac A. , Kiefel V. , Mueller‐Eckhardt C.
Published: March 2003

BACKGROUND: As clinical diagnosis of heparin‐associated thrombocytopenia (HAT) is often difficult, confirmation by sensitive laboratory assays is desirable.

STUDY DESIGN AND METHODS: The sensitivity of the heparin‐induced platelet activation (HIPA) test and the platelet aggregation test (PAT) was prospectively compared by using the sera of 209 patients with the putative diagnosis of HAT. Both assays were performed concomitantly with platelets of the same four donors using a different combination of donors from day to day. Further, all sera were assessed with a platelet factor 4 (PF4)/heparin enzyme‐linked immunosorbent assay (ELISA).

RESULTS: Positive results were obtained with 33 percent of sera in the PF4/heparin ELISA, with 33.5 percent of sera in the HIPA test, and with 11.5 percent of sera in the PAT. The PF4/heparin ELISA and the HIPA test showed no difference in sensitivity (p = 0.27 by McNemar's test) and were more sensitive than PAT (p < 10(‐8) by McNemar's test). However, they recognized different patient cohorts. Nine HIPA‐indeterminate and 12 HIPA‐negative sera were positive in the PF4/heparin ELISA. Eight of the nine indeterminate sera caused platelet activation with high heparin concentrations in the HIPA test. Eleven of the 12 negative sera contained no IgG, but 9 contained IgM and 2 contained IgA HAT antibodies. Four sera that were indeterminate in the PF4/heparin ELISA and 18 sera that were negative were positive in the HIPA test. None of the sera that were positive in the PAT was missed in the HIPA test, but two of those were negative in the PF4/heparin ELISA. All sera were assessed with four low‐molecular‐weight heparins and a low‐molecular‐ weight heparinoid in the HIPA test with platelets from the same four donors. Low‐molecular‐weight heparin caused platelet activation with positive sera in 98 percent of tests, and the heparinoid did so in 10 percent; in a further 12.8 percent, crossreactivity to the low‐ molecular‐weight heparinoid could not be excluded.

CONCLUSION: The majority of HAT antibodies react with a PF4/heparin complex, but there is strong evidence that other antigens are involved in some patients. The HIPA test and the PF4/heparin ELISA are sensitive for diagnosing HAT, and they complement one another.
Circannual variation in lymphocyte subsets, revisited
Authors: Paglieroni T.G. , Holland P.V.
Published: March 2003

Background: Circadian and circannual variations in lymphocyte subsets, especially CD8+ T‐lymphocytes, have been reported. This study focuses on CD4+ T‐lymphocyte seasonal variation over a 6‐year 8‐month period.

Study Design and Methods: Lymphocyte subsets were quantitated monthly for four healthy individuals from 1986 through 1992 as part of a flow cytometry quality‐control program.

Results: In general, there were no significant seasonal changes in the total number of white cells or in total lymphocyte counts. The absolute numbers of CD4+ T‐lymphocytes were lowest in summer when the CD8+ T‐lymphocytes were highest. Mean CD4+ T‐lymphocyte counts were 846, 967, 618, and 695 per microL for Subjects 1 through 4, respectively, in winter and 432, 670, 355, and 766 per microL, respectively, in summer. Two healthy subjects had CD4+ T‐lymphocyte counts lower than 300 per microL on one or more occasions during the study period. In three of the four subjects, the percentage of B‐lymphocytes in winter was almost double that in summer. In one of the four subjects, no circannual rhythm was observed in these lymphocyte subpopulations.

Conclusion: The seasonal variation in CD4+ T‐ lymphocyte counts demonstrated in three healthy individuals over almost 7 years is again of interest in light of renewed consideration of using surrogate tests, such as CD4+ T‐lymphocyte counts, to screen for AIDS‐ like diseases that may be in the blood supply.
Lack of alloimmunization to D antigen in D‐negative immunosuppressed liver transplant recipients
Authors: Casanueva M. , Valdes M.D. , Ribera M.C.
Published: March 2003

BACKGROUND: Orthotopic liver transplantation (OLT) sometimes requires large amounts of blood. An adequate supply of Rh‐negative blood for Rh‐ negative patients is not always available.

STUDY DESIGN AND METHODS: Seventeen Rh‐negative patients, out of 327 receiving OLT in this hospital, received from 5 to 41 units of Rh‐positive red cells during surgery. Each of the 17 patients was followed for 7 weeks to 70 months after OLT for detection of unexpected antibodies. Cyclosporin A and prednisone, azathioprine, and adjunctive rabbit antilymphocyte globulin or monoclonal OKT3 antibody were used to prevent graft rejection.

RESULTS: Evidence of immunization, either to D or to antigens in the rest of the red cell antigen systems, did not appear in any patients.

CONCLUSION: It is hypothesized that cyclosporin A affects the immune humoral response, inhibiting lymphocyte activation and the primary immune response; consequently, Rh‐positive blood may be transfused to Rh‐negative OLT recipients so treated with little or no risk of alloimmunization.
The risk of alloimmunization to c (Rh4) in R1R1 patients who present with anti‐E
Authors: SHIREY R.S. , EDWARDS R.E. , NESS P.M.
Published: March 2003

BACKGROUND: Because the Rh antigens E (Rh3) and c (Rh4) are relatively immunogenic, it has been suggested that R1R1 (E‐, c‐) patients who present with anti‐E alone receive prophylactic c‐ (Rh: −4) red cell transfusions.

STUDY DESIGN AND METHODS: To determine the utility of this approach, the transfusion records of 100 consecutive R1R1 patients with anti‐E identified over a 6‐year period were reviewed.

RESULTS: Thirty‐two (32%) had anti‐c concurrent with anti‐E. Twenty‐seven of the 68 patients who presented with anti‐E alone received random (i.e., not typed for c [Rh4]) red cell transfusions. Five (18.5%) of the 27 subsequently developed anti‐c 13 to 193 days (mean, 50) after transfusion of 2 to 14 (mean, 8) red cell units. None of the five had clinical evidence of hemolysis that could be attributed to a delayed hemolytic transfusion reaction. Twenty‐two (81.5%) of the 27 failed to develop anti‐c even after transfusion of 1 to 41 (mean, 9; median, 7) red cell units.

CONCLUSION: The overall rate of immunization to c (Rh4) antigen in R1R1 patients with anti‐E was 37 percent. Production of anti‐ c following transfusion to R1R1 patients with anti‐E occurred in 18.5 percent of the cases in this series, which could have been avoided by the prophylactic use of R1R1 (E‐, c‐) blood for transfusion. The prophylactic use of c‐ (Rh: −4) blood in this patient population may be justified by the high immunization rate and the potential risk of delayed hemolytic transfusion reaction.
Flow cytometric analysis in platelet crossmatching using a platelet suspension immunofluorescence test
Authors: Skogen B. , Christiansen D. , Husebekk A.
Published: March 2003

BACKGROUND: The sensitivity of flow cytometric measurement of platelet antibodies in a crossmatch technique was investigated.

STUDY DESIGN AND METHODS: The corrected count increment after platelet transfusion was compared with the fluorescence ratio determined by flow cytometric measurement.

RESULTS: When crossmatching was performed before transfusion(s) in alloimmunized patients, a fluorescence ratio < or = 1.7 was associated with satisfactory responses (corrected count increment > or = 7.5), and the predictive values for negative and positive crossmatch results were 94 and 87 percent, respectively. Analysis of antigen preservation during platelet storage with antibodies to HLA alpha‐chain, HLA‐B27, HPA‐1a, and HPA‐3a showed that platelets can be stored, refrigerated, for up to 4 weeks without significant loss of HLA class I and HPA‐1a. There was a slight but continuous loss of HPA‐3a upon storage.

CONCLUSION: Flow cytometric measurement of fluorescence in the platelet suspension immunofluorescence test can be used for platelet crossmatching, with sensitivity and predictive values comparable to those of previously described techniques and with the advantage of automation.
Characterization of reactions after exclusive transfusion of white cell‐ reduced cellular blood components
Authors: Dzieczkowski Jeffery S. , Barrett Barbara B. , Nester Deborah A. , Campbell Mary E. , Cook Jane A. , Sugrue Michele W. , Andersen Janet W. , Anderson Kenneth C.
Published: March 2003

BACKGROUND: Potential adverse effects of white cells (WBCs) within transfused cellular blood components include febrile nonhemolytic transfusion reactions (FNHTRs), alloimmunization, transmission of infectious diseases, transfusion‐related acute lung injury, and immunomodulation. Although exclusive use of WBC‐reduced components to prevent alloimmunization and cytomegalovirus transmission has been studied, the use of these components to avert FNHTR has not been examined. STUDY DESIGN AND METHODS: Transfusion reactions (FNHTRs, allergic reactions, and others) were characterized in recipients of 12,277 WBC‐reduced single‐donor apheresis platelets (SDAPs) and/or red cells (RBCs). Medical and laboratory evaluations for possible infectious and immunologic (alloimmunization) causes of each reaction were undertaken, and the benefit of further modification of components for the prevention of subsequent reactions was also evaluated. RESULTS: Transfusion reactions occurred after 481 (3.92%) of 12,277 transfusions. Allergic reactions occurred more commonly after transfusion of SDAPs (3.69%) than of RBCs (0.51%). Conversely, FNHTRs occurred more commonly after transfusion of RBCs (2.15%) than of SDAPs (1.58%). HLA antibodies were present in a posttransfusion sample from 27 (10.6%) of 255 patients; bacterial contamination was a possible cause of only 2 (0.42%) of 481 reactions. In patients with recurrent FNHTRs, further WBC reduction in components did not wholly prevent further FNHTRs. CONCLUSION: The incidence of FNHTRs and alloimmunization after exclusive transfusion of WBC‐reduced RBCs and SDAPs was low. Further WBC reduction in components transfused to patients with a history of recurrent FNHTRs does not completely prevent subsequent reactions.
Collection of mobilized blood progenitor cells for hematopoietic rescue by large‐volume leukapheresis
Authors: Comenzo Raymond L. , Vosburgh Evan , Weintraub Lewis , Tansan Sualp , Arkin Charles F. , Wright Daniel G.
Published: March 2003

BACKGROUND: Mobilized blood progenitor cells rapidly reconstitute hematopoiesis in patients after dose‐intensive chemotherapy. However, optimal timing and methods of mobilized blood progenitor cell collection have yet to be fully defined. STUDY DESIGN AND METHODS: The utility of large‐volume leukapheresis (LVL; > 15 L blood processed) in collecting target doses of mononuclear cells (7 × 10(8)/kg) for use in autologous hematopoietic rescue was investigated. LVL was begun at a standardized interval (14 days) after a course of limited chemotherapy and subsequent daily recombinant human granulocyte‐macrophage‐colony‐ stimulating factor administration to mobilize blood progenitor cells into the circulation. With each LVL procedure, mononuclear cells, colony‐forming units‐granulocyte‐macrophage (CFU‐GM), burst‐forming units‐erythroid, mixed colonies, total clonogenic progenitor cells, and CD34+ cells collected per kg of patient weight were counted. After high‐ dose chemotherapy and infusion of cryopreserved mobilized blood progenitor cells, the days needed for neutrophils to reach levels of > 0.5 × 10(9) per L and for platelets to reach levels of > 20 × 10(9) per L were recorded. RESULTS: In 14 previously treated cancer patients, an average of 28.9 +/− 4.9 L of blood was processed per LVL (n = 35) to collect medians of 2.5 × 10(8) mononuclear cells per kg (range, 1.0‐ 7.4), 14 × 10(4) CFU‐GM per kg (0‐208), 27 × 10(4) clonogenic progenitor cells per kg (0‐370), and 2.8 × 10(6) CD34+ cells per kg (0‐ 112.5). Fifty‐seven percent of patients (8/14) required one or two LVL procedures to collect adequate blood progenitor cells (range, 1–4). After dose‐intensive chemotherapy, 13 patients received medians of 6.8 × 10(8) mononuclear cells per kg (range, 5.1‐9.9), 53 × 10(4) CFU‐GM per kg (9‐208), and 12 × 10(6) CD34+ cells per kg (3.6‐112.5). Rapid hematopoietic reconstitution occurred with 10 days (range, 8–12) and 9 days (6‐15), respectively, for neutrophil and platelet recoveries. CONCLUSION: Scheduled LVL, beginning on Day 14 after the administration of granulocyte‐macrophage‐colony‐stimulating factor following chemotherapy, is a convenient and efficient method of collecting blood progenitor cells. The mononuclear cells so obtained effected consistent and rapid hematopoietic reconstitution in a highly reproducible manner in a group of heavily treated patients.
Prophylactic versus therapeutic platelet transfusion practices in hematology and/or oncology patients
Authors: Pisciotto Patricia T. , Benson K. , Hume Heather , Glassman Armand B. , Oberman Harold , Popovsky Mark , Hines Deanna , Anderson Kenneth
Published: March 2003

BACKGROUND: Platelet utilization has steadily increased throughout the past three decades. At the same time, there has been very little study of the current transfusion practices. STUDY DESIGN AND METHODS: A survey was conducted of institutional members of the American Association of Blood Banks (hospitals) that were actively involved in the care of pediatric and/or adult hematology and/or oncology patients. Inquiries were made relating to the extent of prophylactic versus therapeutic use of platelets, criteria used for prophylactic transfusion of platelets and type, and dose of platelets used. Data were analyzed according to patient age and type of hospital. RESULTS: Of 786 responding hospitals, 630 (80.2%) provided sufficient data for analysis; 126 of that 630 provided care for pediatric patients. The majority (60.9%) of responding hospitals had a minimum of four hematologists and/or oncologists. Eighty‐four percent of hospitals reported transfusing some apheresis platelets. The dose of platelet concentrates most frequently used for adults ranged from 6 to 10, with pools of 10 more commonly used in community hospitals. More than 70 percent of hospitals reported transfusing platelets primarily for prophylaxis: 60 percent of hospitals set the threshold platelet count for prophylactic platelet transfusion at 20,000 per microL, with approximately 20 percent each transfusing at higher and lower levels. A platelet count of 50,000 per microL was most frequently required for performance of a minor invasive procedure. CONCLUSION: The data from this study show that the majority of institutions use prophylactic platelet transfusion in both pediatric and adult hematology and/or oncology patients. However, there is considerable variation in platelet transfusion practice.
Vascular erosion caused by a double‐lumen central venous catheter during therapeutic plasma exchange
Authors: Quillen Karen , Magarace Lisa , Flanagan Janice , Berkman Eugene M.
Published: March 2003

BACKGROUND: The use of large‐bore double‐lumen dialysis catheters has simplified the procedure of therapeutic plasma exchange, but these catheters are associated with unusual and possibly life‐threatening complications. CASE REPORT: A 46‐year‐old black man was admitted to the hospital with acute onset of paresthesia and weakness. A diagnosis of Guillain‐Barre syndrome was made. Plasma exchange therapy was instituted by peripheral venous access. After three such exchanges, a double‐lumen central venous catheter was placed via the left subclavian vein on hospital Day 7. The patient experienced a sudden onset of severe chest pain and dyspnea during the fourth plasma exchange. He became diaphoretic and hypotensive and experienced tachycardia. The apheresis procedure was stopped. Because of worsening respiratory distress, endotracheal intubation was performed. A chest x‐ray revealed a large right pleural effusion. The central venous catheter was removed. A chest tube was placed, and a large amount of bloody fluid was drained. Several days later, the endotracheal and chest tubes were removed. CONCLUSION: An unusual complication of the use of a central venous catheter, erosion of the superior vena cava, occurred during therapeutic plasma exchange. Prompt recognition of this complication and appropriate therapy can be life‐saving.
Virus inactivation by pepsin treatment at pH 4 of IgG solutions: factors affecting the rate of virus inactivation
Authors: Omar A. , Kempf C. , Immelmann A. , Rentsch M. , Morgenthaler J.‐J.
Published: March 2003

Background: IgG preparations have rarely transmitted infectious diseases; however, because such transmission has occurred a few times, manufacturers are required to present experimental proof that their specific production process removes and/or inactivates viruses that may be present in the starting material.

Study Design and Methods: The kinetics of virus inactivation mediated by pepsin treatment at pH 4 during the production of intravenous immunoglobulin was assessed with spiking experiments using human immunodeficiency virus, bovine viral diarrhea virus, Semliki Forest virus, and pseudorabies virus. The influence of various factors on the rate of virus inactivation also was studied by modifying the composition of the IgG solutions with respect to IgG, sucrose, and NaCl content.

Results: Virus inactivation at 37° C was extremely rapid and resulted in a complete loss of infectivity within 5 minutes to 1 hour. Inactivation was much slower at lower temperatures. Furthermore, inactivation was dependent on the solute composition. Increasing the sucrose content from 0 to 15 percent reduced the rate of inactivation of pseudorabies virus but did not affect the rate of inactivation of Semliki Forest virus. In contrast, increasing the NaCl content from 0 to 150 mM resulted in a reduction in the rate of inactivation of Semliki Forest virus, whereas the rate of inactivation of pseudorabies virus remained unaffected. Moreover, increasing the IgG concentration from 0 to 10 percent resulted in an increased rate of inactivation of pseudorabies virus but a decreased rate of inactivation of Semliki Forest virus.

Conclusion: Inactivation of viruses by pepsin treatment at pH 4 essentially is temperature‐dependent, and the reaction rate is selectively influenced by the solute composition of the IgG solution. This has to be taken into account when safety data for different products are compared.
AS‐1 red cells for neonatal transfusions: a randomized trial assessing donor exposure and safety
Authors: Strauss R.G. , Burmeister L.F. , Johnson K. , James T. , Miller J. , Cordle D.G. , Bell E.F. , Ludwig G.A.
Published: March 2003

Background: Despite recent optimism about the use of erythropoietin therapy to treat the anemia of prematurity, very‐low‐birth‐weight infants who are severely ill receive multiple red cell (RBC) transfusions. Many physicians transfuse relatively fresh RBCs to newborn infants, exposing them to multiple donors and possibly increasing their risk of acquiring transfusion‐transmitted infections.

Study Design and Methods: A randomized, single‐blind clinical trial was conducted to determine, as the primary endpoint, whether RBCs collected from one dedicated donor and stored for ≤ 42 days in AS‐1 storage media could safely supply all small‐volume RBC transfusions (15 mL/kg/dose) needed by very‐low‐birth‐weight infants (0.6‐1.3 kg) during the first 84 days of life. Secondary endpoints were the assessment of the possible adverse clinical and biochemical effects of transfusing AS‐1 RBCs stored for ≤ 42 days. Control infants received identical nursery care, except they received fresh RBCs stored ≤ 7 days in CPDA‐1.

Results: Infants transfused with AS‐1 RBCs were exposed to a mean of 1.6 donors,‐compared with an exposure to 3.7 donors for infants given CPDA‐1 RBCs (p < 0.05). Neither clinical transfusion reactions nor the results of multiple laboratory tests were significantly different in infants who received slow transfusions (15 mL/kg) of AS‐1 RBCs stored for ≤ 42 days and in infants who received the same volume of CPDA‐1 RBCs stored ≤ 7 days.

Conclusion: AS‐1 RBCs, usually from only one dedicated donor, can safely supply all RBCs needed by most very‐low‐birth‐weight infants—a practice that decreases donor exposure and likely increases transfusion safety.
Evaluation of 4,4′‐diisothiocyanatostilbene‐2,2′‐disulfonic acid in the inhibition of rouleaux formation
Authors: Norris S. S. , Allen D. D. , Neff T. P. , Wilkinson S. L.
Published: March 2003

BACKGROUND: DIDS (4,4′‐diisothiocyanatostilbene‐2,2′‐disulfonic acid) inhibits the formation of serum‐or plasma‐induced rouleaux through its ability to bind to band 3 on red cell membranes. This property of DIDS was evaluated in the serologic testing of specimens exhibiting rouleaux.

STUDY DESIGN and METHODS: Optimal test conditions for DIDS treatment of reagent red cells were determined by varying the volume and concentration of DIDS solution and the incubation temperature and duration and comparing the results of antibody screening procedures using specimens that exhibit macroscopically visible rouleaux. Blind titration studies compared untreated and DIDS‐treated red cells to evaluate the maintenance of antigen integrity. The use of DIDS‐treated red cells for antibody detection and identification was evaluated by comparing the results in donor specimens containing antibodies with those in untreated and DIDS‐treated selected panel cells. In addition, 4‐percent (wt/vol) dextran in serum was used to induce rouleaux formation as a way of determining the capability of DIDS to resolve ABO serum grouping discrepancies.

RESULTS: Complete inhibition of rouleaux formation occurred when reagent red cells were treated by incubation at 37 degrees C for 10 minutes with 150 microliter (approx. 5 drops) of 0.05 mg per mL of DIDS in 0.9‐percent NaCl. There were no significant differences in titration scores of untreated and DIDS‐treated red cells tested with the 19 antisera used to assess antigen integrity. Antibody identification studies showed that DIDS‐treated reagent red cells reacted similarly to untreated reagent red cells. In addition, DIDS resolved dextran‐induced ABO serum grouping discrepancies.

CONCLUSION: DIDS effectively resolved the serologic problems associated with the presence of rouleaux, without affecting the results of the test system itself. Implementation of this method to inhibit the rouleaux‐forming properties of serum and plasma specimens can be useful in serologic testing.
Posttransfusion graft‐versus‐host disease in Japanese newborns
Authors: Ohto H. , Anderson K. C.
Published: March 2003

BACKGROUND: Posttransfusion graft‐versus‐host disease (PT‐GVHD) is underdiagnosed and underreported. Risk factors predisposing to PT‐GVHD in newborn recipients, the clinical manifestations of the disease in newborns, and its mechanism are not well characterized.

STUDY DESIGN and METHODS: A literature review identified 27 cases of PT‐GVHD in newborns in Japan. Detailed information on volume of blood transfused, donor(s), and clinical course was analyzed. Infants with known immunodeficiency were excluded.

RESULTS: Of 27 newborns, 20 were premature and 7 were full‐term. Thirteen premature neonates were transfused frequently because of anemia; in 10 cases, exchange transfusion was performed. Fresh blood (<or = 72 hours after donation) and blood from family member(s) were transfused to 25 and 22 neonates, respectively. The clinical manifestations of PT‐GVHD developed later after transfusion than is reported in adults; the median intervals were: fever, 28 days; erythematous rash, 30 days; leukopenia, 43 days; and death, 51 days. Thymic damage was a striking feature among newborns. Two thirds of the cases of PT‐GVHD were associated with overwhelming infections, and all patients died in spite of various treatments.

CONCLUSION: These data suggest that cellular blood components, especially those from family members, should be irradiated before transfusion to premature and full‐term neonates. Extrathymic and/or thymic semitolerance for allogeneic‐donor cytotoxic T‐ lymphocytes may explain the longer period of latency before the onset of clinical manifestations and the longer course of disease before death in newborns than are noted in adults, as well as the fewer than expected reported cases of PT‐GVHD in neonates.
Chloramphenicol‐dependent antibody: a case report
Authors: Fried M. R. , Scofield T. L. , Stroncek D. F. , Swanson J. L.
Published: March 2003

BACKGROUND: Chloramphenicol‐dependent antibodies are a rare cause of interference in pretransfusion serologic testing. Their presence can be confirmed by the testing of red cells in both the presence and absence of chloramphenicol.

CASE REPORT: A 29‐year‐old, group A, Rh‐positive man with no history of chloramphenicol exposure was found to have a chloramphenicol‐dependent panagglutinin in his serum. The antibody was IgM with a titer of 8. It showed no blood group specificity when tested with common and rare red cell phenotypes, and it failed to react with platelets and granulocytes. Confirmation attempts using a chloramphenicol sodium succinate solution as the cell‐suspending medium led to negative results. The antibody reacted serologically only in the presence of chloramphenicol, which arises from the succinate derivative by the action of blood esterases.

CONCLUSION: This case is an additional example of a chloramphenicol‐dependent antibody. It demonstrates how the laboratory investigation of drug‐related phenomena is dependent on testing the drug from that reacts in vivo.
An automated system for bedside verification of the match between patient identification and blood unit identification
Authors: Jensen N.J. , Crosson J.T.
Published: March 2003

Background: The administration of blood to the wrong patient remains the leading cause of acute hemolytic transfusion reactions and subsequent death. A process control system for blood administration was developed that verifies, at the bedside, the match between barcoded patient identification and blood unit identification.

Study Design and Methods: The system is composed of 1) a portable bedside scanner that reads barcoded patient identification and blood unit identification, 2) a host computer system capable of accepting transfusion data from the bedside scanner, 3) printed documentation of the transfusion episode, and 4) audit trail monitoring of whether all steps in the automated patient and blood unit identification process have been performed. Software design, development, and validation protocols followed industry standards.

Results: A pilot study was performed over a 2‐month period evaluating the blood administration process using the computerized bedside transfusion identification system prototype for transfusions in 39 oncology patients.

Conclusion: This system controls the blood administration process and includes bedside verification of the match between patient identification and blood unit identification.
Determination of sibship in any two persons
Authors: Wenk R.E. , Traver M. , Chiafari F.A.
Published: March 2003

Background: Parentage testing laboratories may be asked to provide genetic evidence that two persons are or are not related, when no other relatives are available for study. Simple methods using autosomal, codominant, unlinked genetic systems can determine if two people are blood relatives (e.g., siblings).

Study Design and Methods: The odds ratios (full sibship index) of true sibling pairs were determined from two‐child paternity cases and compared with regionally and racially matched control pairs of unrelated children. The sharing of two, one, or no alleles was observed in pairs of children at three independent, polymorphic VNTR (variable number of tandem repeat) sequences loci. The sibship index was calculated as (the chance that an observation would occur if two children were siblings) divided by (the chance that it would occur if the two were unrelated). Sibship indices and the frequencies of shared alleles were determined for 20 sibling pairs and 20 control pairs.

Results: Sibship index values were less than 1 in all 20 pairs of unrelated children. Sibship index values were greater than 100 in nine pairs of siblings (45%), between 10 and 100 in five pairs (25%), between 1 and 10 in four pairs (20%), and less than 1 in two pairs (10%). Siblings shared two alleles in 17 of 60 observations (28.3%); controls shared two alleles in 0 of 60 observations (0%).

Conclusion: The sharing of one allele and the sharing of no alleles at a polymorphic locus of high heterozygosity provide limited information for and against sibship, respectively. The sharing of two alleles produces strong evidence favoring sibship. In a given case, the study of more than three polymorphic loci of high heterozygosity may be needed to develop the evidence that two people are siblings. The general logic and methods used for siblings apply to kinship analyses of other two‐person pedigrees.
Comparison of the polyethylene glycol antiglobulin test and the use of enzymes in antibody detection and identification
Authors: Reisner R. K. , Butler G. W. , Bundy K. L. , Moore S. B.
Published: March 2003

BACKGROUND: The polyethylene glycol indirect antiglobulin test for detection of red cell antibodies was compared with a proven, highly sensitive test system using papain. STUDY DESIGN AND METHODS: Parallel, prospective testing of 1508 samples with polyethylene glycol and with albumin and papain evaluated the sensitivity and specificity of polyethylene glycol. Retrospective analysis of antibody specificities was performed for the 2 years before and the 2 years after the institution of polyethylene glycol testing. RESULTS: Of 1508 prospective screens, 53 (3.5%) had discordant results: 5 were positive only in polyethylene glycol and 48 were positive only in albumin and papain. Upon antibody identification, the 5 samples that were positive only in polyethylene glycol showed 1 anti‐D, 2 warm autoantibodies, and 2 false‐positive results. The 48 samples that were positive only in albumin and papain showed 1 each of the following: anti‐Le(b); anti‐P1; anti‐S; high‐titer, low‐avidity antibody; and cold autoantibody; there were 43 false‐positive results. False‐positive results totaled 12 (0.8%) with polyethylene glycol and 53 (3.5%) with albumin and papain. The retrospective analysis of antibody specificity with polyethylene glycol showed a significant increase in the detection of Fy(a) and/or Fy(b) (p < 0.0002) and Jk(b) (p < 0.0002) antibodies and a decrease in the detection of Le(a) and/or Le(b) antibodies (p < 0.0002). CONCLUSION: Polyethylene glycol retained the high sensitivity of the albumin and papain, while significantly lowering the number of false‐ positive results and decreasing the detection of antibodies of doubtful clinical significance.
Point mutations characterize KEL10, the KEL3, KEL4, and KEL21 alleles, and the KEL17 and KEL11 alleles
Authors: Lee S. , Wu X. , Son S. , Naime D. , Reid M. , Okubo Y. , Sistonen P. , Redman C.
Published: March 2003

BACKGROUND: The Kell blood group system is complex, consisting of five sets of alleles and expressing high‐ and low‐incidence antigens and at least 11 other independently expressed antigens. The molecular basis of two sets of alleles: KEL1 (K) and KEL2 (k) and KEL6 (Jsa) and KEL7 (Jsb) have been elucidated as single‐base mutations leading to amino acid changes. The molecular basis for the KEL3 (Kpa), KEL4 (Kpb), and KEL21 (Kpc) alleles, the KEL11(Cote) and KEL17(Wka) alleles, and for KEL10 (UIa) is now reported. STUDY DESIGN AND METHODS: Genomic DNA from unrelated individuals with KEL:3,‐4,‐21 [Kp(a+b‐c‐)], KEL:‐3,‐4,21 [Kp(a‐b‐c+)], KEL:17,‐11, and KEL:10 (UIa) phenotypes was amplified by polymerase chain reaction (PCR) with primers for the 19 exons of KEL. The PCR products were sequenced and compared to the DNA sequences of a common Kell system phenotype, KEL:‐3,4,‐21,‐17,‐10. Base mutations found were confirmed by restriction fragment length polymorphism analysis in which DNA of unrelated persons with similar red cell phenotypes was used. RESULTS: In all cases, single‐base mutations were responsible for the expression of the various antigens. In KEL3 (Kpa), KEL4 (Kpb), and KEL21 (Kpc), point mutations at the same codon in exon 8, encoding amino acid residue 281, distinguish the three genes. KEL4 has the CGG codon for arginine, KEL3 has the TGG codon for tryptophan, and KEL21 has the CAG codon for glutamine. KEL17 has a T1025C mutation in exon 8, encoding a valine‐to‐alanine amino acid change at residue 302. KEL10 has an A1601T mutation in exon 13, encoding a glutamic acid‐ to‐valine change at residue 494. In all cases, the point mutations created restriction enzyme sites, and PCR‐based restriction fragment length polymorphisms confirmed that these point mutations occurred in unrelated persons with the same red cell phenotype. CONCLUSION: Single‐ base substitutions characterize the KEL3, KEL21, KEL17, and KEL10 genes. The allelic relationship of KEL3, KEL4, and KEL21 was confirmed because the mutations occur in the same codon, expressing different amino acids. PCR‐based restriction fragment length polymorphisms can be used to distinguish genotypes.
Genotyping of KEL1 and KEL2 of the human Kell blood group system by the polymerase chain reaction with sequence‐specific primers
Authors: Hessner M. J. , McFarland J. G. , Endean D. J.
Published: March 2003

BACKGROUND: Kell is a major antigenic system in human red cells, with more than 20 identified antigens. KEL1 and KEL2 are two opposing low‐ and high‐frequency alleles. Immunization to KEL1 is clinically significant, because anti‐KEL1 can cause severe reactions to transfusion of incompatible blood, as well as hemolytic disease of the newborn. At the nucleotide level, the difference between the KEL2 and KEL1 alleles is a single‐base change within exon 6 that results in the substitution of methionine (ATG) for threonine (ACG) at position 193. STUDY DESIGN AND METHODS: An assay using polymerase chain reaction and sequence‐specific primers to genotype for the KEL1 and KEL2 alleles has been developed. It uses two allele‐specific forward primers for either KEL1 or KEL2 and a single reverse‐consensus primer. RESULTS: A validation study of 42 serologically typed samples (5 KEL:1,‐2 [K+k‐]; 23 KEL:1,2 [K+k+]; and 14 KEL:‐1,2 [K‐k+]) was performed. A concordance rate of 100 percent (42/42 samples) was observed between polymerase chain reaction with sequence‐specific primers and serologic typing. CONCLUSION: This rapid, nonradioactive, Kell system genotyping assay does not require the additional steps of probe hybridization or restriction enzyme digestion. This application of polymerase chain reaction with sequence‐specific primers should prove particularly useful in Kell system genotyping of amniotic cells to identify pregnancies at risk for hemolytic disease of the newborn.
Involvement of Ser103 of the Rh polypeptides in G epitope formation
Authors: Faas B. H. W. , Beckers E. A. M. , Simsek S. , Overbeeke M. A. M. , Pepper R. , van Rhenen D. J. , von dem Borne A. E. G.Kr. , van der Schoot C. E.
Published: March 2003

BACKGROUND: Almost all red cells that carry D and/or C antigens also express the G antigen (Rh12). A study was conducted on the molecular background of the G epitope. STUDY DESIGN AND METHODS: Two unrelated donors with the rare ccDEe, G‐ phenotype and one donor with the ccEe, G+ phenotype were studied. Genomic DNA and cDNA of these donors were studied with polymerase chain reaction, Southern blot, and sequence analysis, with special focus on exon 2, because it is only in this exon that there are supposed to be similarities between RHD and the RHC allele, but not between RHD and the RHc allele. RESULTS: In both ccDEe, G‐ donors, a nucleotide substitution was found in exon 2 of RHD; T307 was replaced by C307, which predicted a Ser‐>Pro substitution at amino acid position 103 of the D polypeptide. The ccEe, G+ donor carried the complete exon 2 of RHD. Moreover, despite the absence of all known D epitopes, this donor also carried RHD characteristics in exons 1 to 3 and exon 9 and further downstream. CONCLUSION: Ser103, encoded by exon 2 of the RH genes, is involved in G epitope formation.
Characterization of antibodies produced by S‐s‐individuals
Authors: Storry J. R. , Reid M. E.
Published: March 2003

BACKGROUND: Historically, classification of U‐ and U variant (U+var) individuals has been made by hemagglutination and adsorption and elution studies performed with polyclonal U antisera. Molecular studies and serologic tests with a potent monoclonal anti‐He have shown that U+var red cells, some of which are He+, possess an altered form of glycophorin B. STUDY DESIGN AND METHODS: Seventeen sera, previously determined to contain anti‐U, were tested with a panel of red cells of common and rare MNS types. RESULTS: Five sera contained anti‐U only, and 12 sera contained broadly reactive antibodies with apparent, but inseparable, anti‐U,He or anti‐U,N,He specificities. CONCLUSION: The majority of antibodies produced by S‐s‐U‐ individuals are anti‐U plus anti‐glycophorin B and are analogous to the broadly reactive antibodies produced by En(a‐) individuals whose red cells lack glycophorin A or have altered glycophorin A. To avoid further immunization of patients with anti‐U, sera used for classification of S‐s‐U‐ donors should be selected to detect S‐s‐ red cells that possess altered forms of glycophorin B.