摘要:

BACKGROUND: Allogeneic blood cell transplantation is a new alternative to bone marrow transplantation. Preliminary data suggest that granulocyte‐colony‐stimulating factor (G‐CSF)‐mobilized peripheral blood progenitor cells from normal donors can provide rapid hematopoietic engraftment without significant increases in transplant‐ related morbidity. Potential advantages to donors include the elimination of an operation under general anesthesia. STUDY DESIGN AND METHODS: Twenty‐one normal donors underwent high‐dose (16 pg/kg/day for 5 days) G‐CSF mobilization. Apheresis was performed on the fifth and sixth days of G‐CSF administration, and apheresis components were analyzed by flow cytometry. Donor characteristics in relationship to apheresis yields were also analyzed. RESULTS: Apheresis components were analyzed according to donor weight. The median total numbers of white cells per kg, CD34+ cells per kg, and CD3+ cells per kg were 10.8 × 10(6), 7.2 × 10(8), and 295 × 10(8), respectively. Day 5 collections had significantly higher nucleated cell content and median CD34+ percentages than did Day 6 collections (0.71% on Day 5 vs. 0.58% on Day 6, p<0.01). CD34+ content and total white cells were not related to age or sex. No donors experienced toxic effects that required their removal from the study. CONCLUSION: Day 5 is the optimal day for the harvest of normal‐donor peripheral blood progenitor cells after mobilization with high‐dose G‐CSF. Older individuals are acceptable donors of a

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091734.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: Granulocyte‐colony‐stimulating factor (G‐CSF) is used for the mobilization of progenitor cells and granulocytes. False‐positive hepatitis B surface antigen (HBsAg) enzyme‐linked immunosorbent assays (ELISAs) (NML) from one manufacturer in individuals receiving G‐CSF have been observed. STUDY DESIGN AND METHODS: Sixty‐six autologous peripheral blood progenitor cell donors from 1994 were retrospectively reviewed. Donors typically received 5 to 10 micrograms of G‐CSF per kg subcutaneously for 5 days before collection. Additional ELISA dilutional studies (1‐in‐10, 1‐in‐100, 1‐in‐1000) with known HBsAg‐ negative serum were made with G‐CSF. Testing was performed by the University of North Carolina, the American Red Cross in Charlotte, NC, or the National American Red Cross, Washington, DC. RESULTS: Of the 66 patients, none reacted for antibody to hepatitis B core antigen, and 30 (45%) had a positive reaction in the ELISA. Surface antigen positivity was “confirmed” on 6 of the 30 patients by neutralizing ELISA reactivity with an antibody to HBsAg test from the same manufacturer. In all cases, the clinical presentation was not suggestive of hepatitis, and these individuals were not at high risk for hepatitis B. Twenty‐seven of the 30 cases were tested with a monoclonal HBsAg ELISA (AUSZYME) from another manufacturer in the peridonation period and did not react. In 1994, 256 autologous whole‐blood donors not receiving G‐ CSF were similarly tested and only 1 (0.4%) had a positive reaction with the second manufacturer's HBsAg ELISA (p<0.001). Of this group, 41 patients with histories of malignancy were identified, which is comparable to the history of the peripheral blood progenitor cell donors in this study, and none of these blood donors tested positive for HBsAg (p<0.001). Dilutional studies with G‐CSF produced no reactions. CONCLUSION: The NML HBsAg ELISA studied has an unacceptably high false‐positive rate in patients or donors receiving G‐CSF. The false reactivity of this assay appears to be an indirect consequence of G‐CSF administration, which can also lead to spurious confirmation by the HBsA

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091735.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: Fever has been identified as a major cause of platelet refractoriness. However, it is regularly attended by the administration of antibiotics, and thus it cannot be determined whether fever, the administration of antibiotics, or both are responsible for the reduced platelet increment. The present study was undertaken to discern the effect of body temperature and antibiotics on transfusion success. STUDY DESIGN AND METHODS: Single‐donor platelet transfusions (n = 400) were monitored retrospectively. Besides other influencing factors (e.g., spleen size, number of previous transfusions, diagnosis, bone marrow transplantation), body temperature and the administration of antibiotics were documented from the patients' records. To distinguish the effects of fever and antibiotics, a general mixed model of variance was used for analysis. RESULTS: Besides the well‐known factors of splenomegaly, hepatomegaly, bone marrow transplantation, and pretransfusion storage time, both body temperature and antibiotics independently reduced the corrected count increment. Among the various antibiotics, amphotericin B, ciprofloxacin, and vancomycin had the greatest influence. CONCLUSION: The administration of antibiotics has a negative effect on the corrected count increment, independent of the presence of fever. Besides amphotericin B, which has previously been shown to influence the corrected count increment, vancomycin and ciprofloxacin reduce it significantly. Because these antibiotics are widely used in patients with bone marrow failure, these observations should be proven by further prospective in vivo and in vitro stud

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091736.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: The use of fresh platelets results in better posttransfusion recovery and survival than does the use of platelets that have been stored before transfusion. Activation of platelets during preparation and storage may be one of the factors responsible for a number of storage‐related changes in platelet membrane proteins. Blood centers commonly prepare platelet concentrates from both multiple units of whole blood and single‐donor plateletpheresis collections. STUDY DESIGN AND METHODS: Seventeen plateletpheresis concentrates, anticoagulated with ACD, were compared to platelets prepared from whole blood from the same donor that was anticoagulated with CPDA‐1 (random‐ donor platelets). After preparation, plateletpheresis and random‐donor platelets were stored in plastic storage bags at 22 degrees C for 5 days. Platelet surface glycoproteins were examined by flow cytometry after platelets were fixed in dilute plasma with 1‐percent formaldehyde and stained with fluorescein isothiocyanate‐labeled monoclonal antibodies CD42b (anti‐glycoprotein [GP]lb), CD41a (anti‐GPllb/llla), and CD62 (anti‐P‐selectin). RESULTS: The binding of anti‐CD42b was greater in plateletpheresis concentrates than in random‐donor platelets on Days 3 and 5 (p<0.01) of storage; binding of anti‐CD62 was greater in the random‐donor concentrates (p<0.01) on Days 3 and 5. Plateletpheresis concentrate aggregation responses were greater on Day 5 (p<0.01). To determine if the type of anticoagulant and the method of mixture with blood contributed to these changes, 10 samples were split into aliquots and prepared in two separate ways: One group of samples was prepared by allowing anticoagulant (ACD) and blood to flow into the tube at a rate of 3 microL per second, and the other group of samples was prepared by allowing blood to flow into tubes containing a measured amount of CPDA‐1. The first samples bound more anti‐CD42b than the second samples (p<0.01). The second group of samples contained significantly more microvesicles that bound anti‐CD41a than did the first group (p<0.01). Samples prepared by the first method but anticoagulated with CPDA‐1 contained more microvesicles but had the same amount of anti‐CD42b binding as did similarly prepared samples anticoagulated with ACD (p<0.05). CONCLUSION: Platelet concentrates prepared from single units of whole blood and anticoagulated with CPDA‐ 1 bind less anti‐CD42b and more anti‐CD62 than do platelets obtained by apheresis. These differences may be attributed to platelet sedimentation and the transient exposure of some of the platelets in the blood that is first collected during whole‐blood d

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091737.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: The mechanisms of the detrimental effects of perioperative allogeneic blood transfusion are still unclear. Previous studies have suggested a higher incidence of adverse effects after the use of blood stored for prolonged time. Therefore, a possible time‐dependent release of various white cell‐ and platelet‐derived bioactive substances in stored human red cell suspensions was studied. STUDY DESIGN AND METHODS: Whole blood (6 units), plasma‐reduced whole blood (6 units), and saline‐adenine‐glucose‐mannitol blood (6 units) from 18 unpaid, normal blood donors were stored under standard blood bank conditions at 4 degrees C for 35 days. After refrigeration, samples were collected from all blood bags on Days 0, 2, 5, 9, 14, 21, 28, and 35 of storage. Extracellular concentrations of eosinophil cationic protein, eosinophil protein X, plasminogen activator inhibitor 1, myeloperoxidase, and interleukin 6 were analyzed by enzyme‐linked immunosorbent assay and radioimmunoassay. The total intracellular and donor plasma levels of these substances also were analyzed at the time of blood donation. RESULTS: Eosinophil cationic protein, eosinophil protein X, and myeloperoxidase increased 10‐ to 25‐fold (p<0.05) in a time‐dependent manner in whole blood, plasma‐reduced whole blood, and saline‐adenine‐ glucose‐mannitol blood during storage for 35 days. Plasminogen activator inhibitor 1 increased threefold to sixfold (p<0.05) in whole blood and plasma‐reduced whole blood, but not in saline‐adenine‐ glucose‐mannitol blood. Interleukin 6 was not detected in either plasma or samples obtained from the blood bags. CONCLUSION: Stored whole blood, plasma‐reduced whole blood, and saline‐adenine‐glucose‐mannitol blood may release white cell‐ and platelet‐derived bioactive substances in a time‐dependent manner, which may be related to the detrimental effects of perioperative blood transfusions. Therefore, prestorage white cell reduction should be conside

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091738.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: Some of the major blood group antigens are on lipids and proteins of the red cell membrane. Incubation of intact red cells with liposomes containing specially designed artificial lipids has been shown to result in the extraction of membrane proteins by the liposomes. The extraction of blood group structures and the retention of their antigenicity have not been reported. STUDY DESIGN AND METHODS: After the incubation of red cells with liposomes, the extraction of the antigens from human red cells by liposomes was examined by evaluation of the agglutination of the liposomes by respective antisera. RESULTS: Agglutination specific to the A and B blood group antigens was seen, which indicated that the antigenicity of the blood group antigens was retained even after the extraction by the liposomes. The presence of an artificial boundary lipid, 1,2‐dimyristamido‐1,2‐ deoxyphosphatidylcholine, in the liposome was crucial to the efficient extraction of the A and B antigens. On the other hand, the extraction of D, M, N, and P1 was not always detectable by agglutination. CONCLUSION: The A and B blood group antigens were directly extracted from red cells by liposomes without loss of antigen

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091739.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: Screening of blood donors for Chagas' disease by using currently available serologic tests is complicated by the lack of adequate sensitivity, discordant results between tests, and the absence of a gold standard. STUDY DESIGN AND METHODS: The study was designed to evaluate the serologic tests by using epidemiologic data relating to the risk of exposure to Trypanosoma cruzi in the urban centers of Brazil. The serologic results obtained from screening 411,617 voluntary blood donations in Sao Paulo during 1993 and 1994 were reviewed, as well as follow‐up results on 1,267 donors who initially were repeatably reactive in at least one of three screening tests. Epidemiologic data were obtained from 321 individuals who on follow‐up remained reactive in at least one test and who returned for medical counseling. Controls included 119 screen‐negative blood donors and 45 blood donors who were repeatably reactive in at least one screening test but were negative on follow‐up. RESULTS: Of the individuals who reacted in three screening tests, 94.6 percent remained reactive on follow‐up. Of the individuals who were repeatably reactive in only one screening test, 70.8 percent were negative in all three tests on follow‐up. Most individuals who reacted in two or three tests on follow‐up had epidemiologic evidence of a risk of exposure to Chagas' disease. A significant proportion (29.1%) of those who were reactive in only one test on follow‐up had epidemiologic evidence of exposure to the Chagas' disease vector as compared to 14.6 percent of controls (p = 0.007). This suggests that some of these individuals truly were infected. CONCLUSION: No single test for Chagas' disease is sufficiently sensitive to prevent transfusion transmission of the disease in the urban ce

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091740.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: In a recent quality assurance audit of component returns over a 6‐month period, 9 of 81 returns were due to icteric plasma. With the sensitive, new methods used to screen donors for anemia and hepatitis, it seemed likely that the icteric discoloration reflected benign unconjugated hyperbilirubinemia (Gilbert's syndrome) in the donor, rather than liver disease or hemolysis. The donors were recalled for repeat blood study to resolve this question. STUDY DESIGN AND METHODS: Seven of the nine donors could be reached, and they submitted blood samples for measurement of serum levels of conjugated (direct‐ reacting) and total bilirubin and for complete blood and reticulocyte counts. RESULTS: All seven donors had mild unconjugated hyperbilirubinemia, with total bilirubin levels ranging from 1.3 to 2.8 mg per dL. None showed evidence of overt hemolysis. CONCLUSION: All seven donors of the components with icteric plasma have Gilbert's syndrome, a benign genetic anomaly occurring in approximately 3 to 5 percent of the general population. With the sensitive screening tests for viral hepatitis used today, the presence of icteric plasma in a component suggests that the donor has Gilbert's syndrome. Policies about the acceptability of icteric components from blood donors merit reassessm

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091741.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: The association between HLA class II antigens and the production of red cell autoantibodies was investigated in this study. STUDY DESIGN AND METHODS: Thirty‐one individuals with a positive direct antiglobulin test (DAT) and 85. DAT‐negative controls (cadaveric organ donors) were typed using a DNA‐based method for class II HLA typing, that is, the polymerase chain reaction with sequence‐specific primers. Amplified HLA alleles corresponding to the serologic specificities DR 1‐ 18 and DQ 1–9 were identified. RESULTS: Analysis of DR and DQ frequencies showed that HLA‐DQ6 was less frequent in DAT‐positive individuals (19% vs. 53% in the control population), with a p value of 0.0014 and a corrected p value of 0.059, and relative risk of 0.23 (95% Cl = 0.09‐0.58). The frequency of HLA‐DQ6 was higher in asymptomatic DAT‐positive blood donors (n = 8, 38% DQ6 positive, p = 0.48) than in DAT‐positive hospital and clinic patients (n = 23, 13% DQ6 positive, corrected p value = 0.030, relative risk = 0.13, 95% Cl = 0.04‐0.41), 96 percent of whom had evidence of clinical hemolysis. CONCLUSION: This study demonstrates that HLA‐DQ6 may have a negative association with a positive DAT result in patients with evidence for hemolysis, and may be a resistance antigen for clinically relev

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091742.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

BACKGROUND: Flow cytometry is used to quantitate CD34+ hematopoietic progenitor cells for transplantation. The present study evaluates methods for preparing and thawing cryopreserved CD34+ and CD34‐ cell lines for use as flow cytometry reagents. STUDY DESIGN AND METHODS: The human myeloid leukemic cell lines KG 1 a (CD34+) and K562 (CD34‐) were grown in culture under standard conditions and then prepared on ficoll gradients of different densities to determine which gave the component that was most reproducible. After ficoll preparation, the cells were frozen in standard cryopreservation media and four methods of thawing were examined. Determination of the method that gave the cell component that was most reproducible was based on viability, percentage of cell recovery, and maintenance of CD34 antigenicity status. RESULTS: Ficoll gradient preparation improved the ease of flow cytometry analysis when original viability was low, and it produced a more uniform cell population. However, it resulted in significant cell loss for both cell lines. While the cell recovery for K562 cells was not significantly different with any of the densities of ficoll, recovery was significantly better for KG 1 a cells with ficoll at a specific gravity of 1.077. Of the thawing methods examined, all three that involved a rapid thaw at 37 degrees C were statistically equivalent to each other and were better than thawing at 4 degrees C. CONCLUSION: With a standardized method of preparing cell lines as reagents for quality control purposes, data comparison among cell processing laboratories may more readily be initiated. Such cell lines could also be useful as a teaching tool for flow cytometry and in proficiency test

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36111297091743.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY