ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496172062.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:Irradiation of platelet concentrates (PCs) with ultraviolet‐ B (UVB) light inactivates the contaminating white cells and might be an alternative to filtration for the prevention of alloimmunization to HLA antigens and subsequent refractoriness to further platelet transfusions in multiply transfused patients with bone marrow failure.Study Design and Methods:Patients with hematologic malignancy, mainly acute myeloid leukemia, were prospectively assigned in a random manner to receive either UVB‐irradiated or control, nonirradiated PCs. All patients were given red cells that were white cell reduced by filtration. Transfusion efficacy and alloimmunization were assessed by means of corrected count increments, requirement for red cells and PCs, and measurement of lymphocyte‐reactive antibodies.Results:UVB‐irradiated PCs had a clinical efficacy similar to controls as judged by corrected count increments at 1 to 6 and 12 to 24 hours and by the median requirement for red cell and platelet transfusions. Alloimmunization determined by measurements of lymphocyte‐reactive antibodies using both conventional and antiglobulin‐augmented lymphocytotoxicity techniques was not abolished in recipients of UVB‐irradiated PCs (4/30, 13%) but was less than that in controls (5/20, 25%; p = NS). The mean number of platelet transfusion episodes prior to the occurrence of alloimmunization was greater in the control group (27 vs. 10; p = 0.017).Conclusion:In this trial, UVB irradiation did not diminish the clinical efficacy of platelet transfusions. There was a small but nonsignificant reduction alloimmunization, but no difference in refractoriness of the two groups was observed. Larger prospective randomized studies are required to confirm these findings and to compare UVB irradiation with white c

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226140.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:Bone marrow, peripheral blood, and umbilical cord blood have been used to prepare autologous and allogeneic pluripotential mononuclear cells for use in the repopulation of bone marrow.Study Design and Methods:The purpose of this study was to evaluate how the temperature and duration of frozen storage of human peripheral blood mononuclear cells (PBMCs), as well as the freezing container, affected the in vitro recovery and viability of the mononuclear cells and their growth in colony‐forming unit‐granulocytic‐erythroid‐monocytic‐ megakaryocytic (CFU‐GEMM) tissue culture assay. PBMCs were isolated from ficoll‐hypaque‐treated cellular residue obtained during the plateletpheresis of blood from 15 healthy donors. The PBMCs were treated with dimethyl sulfoxide (DMSO) to achieve a final DMSO concentration of 10 percent. Each unit was then separated into six aliquots: one stored in a polyvinylchloride (PVC) plastic bag, one in a polyolefin plastic bag, and four in polyethylene cryostorage vials. Each aliquot was frozen in a ‐80 degrees C mechanical freezer at a freezing rate of 2 to 4 degrees C per minute. The frozen PBMCs in PVC bags were stored in a ‐80 degrees C mechanical freezer and those in polyolefin bags in a ‐135 degrees C mechanical freezer. Each of the four frozen samples in a vial was stored at a different temperature: one in the ‐80 degrees C freezer, one in the ‐135 degrees C freezer, one in the vapor phase of liquid nitrogen at ‐150 degrees C, and one in liquid nitrogen at ‐197 degrees C. Some of the frozen PBMCs were stored for periods of 1 to 1.5 years and others for 2 to 2.4 years, after which they were thawed, washed, and tested.Results:The samples stored in PVC bags and those stored in polyolefin bags exhibited in vitro recoveries that were 90 percent of the recovery of fresh PBMCs and viabilities of 90 percent after 2.4 years of frozen storage. The PBMCs stored in PVC bags exhibited no loss of CFU‐GEMM activity after 1 to 1.5 years, but a 40‐percent loss of activity was observed after 2 to 2.4 years. PBMCs stored in polyolefin bags, however, exhibited no loss of CFU‐GEMM activity, even after 2 to 2.4 years of storage. In vitro recovery was significantly lower in PBMCs stored in vials at ‐80 degrees C or ‐135 degrees C than in cells stored in PVC or polyolefin bags at these temperatures, both in the 1‐ to 1.5‐year and the 2‐ to 2.4‐year time frames. In vitro recovery and viability were similar in PBMCs stored in vials at ‐80 degrees C, ‐135 degrees C, ‐150 degrees C, and ‐197 degrees C. The growth patterns in the CFU‐GEMM assay in PBMCs stored in vials were significantly lower after storage at ‐80 degrees C than after storage at ‐135 degrees C, ‐150 degrees C, or ‐197 degrees C.Conclusion:PBMCs isolated by leukapheresis and ficoll‐hypaque treatment can be frozen with 10‐percent DMSO in a ‐80 degrees C mechanical freezer. When a PVC bag is used for freezing and storage of PBMCs at ‐80 degrees C, the duration of frozen storage should not exceed 1.5 years, whereas PBMCs frozen in a polyolefin bag can be stored in a ‐135 degrees C freezer for as long as 2.4 years. When these guidelines were followed, in vitro recovery was 90 percent that of fresh PBMCs, viability was 90 percent, and growth in the CFU‐GEMM tissue culture assay was similar to that of fresh PBMCs. The PBMCs frozen and stored in PVC or polyolefin bags exhi

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226141.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:The commonABOallele sequences are known, but little or no genetic information is available on the rare but important A subgroups.Study Design and Methods:Blood group ABO polymorphism was analyzed in genomic DNA from 45 rare subgroup A individuals by sequence‐specific primer polymerase chain reaction and amplified fragment length polymorphism investigating exons VI and VII in the ABO genes. These methods are used to detect specific mutations only, and not all changes that might be present can be detected. ABO genotypes discriminating six alleles (A1,A2, B,O1,O1var, andO2) were determined.Results:The C→T substitution at nucleotide position 467 (C467T) is not restricted to A2and cis‐AB individuals, but was found also in some A subgroups. Detection of the functionally more relevant C1060‐single‐point deletion inA2was accomplished by a novel sequence‐specific primer polymerase chain reaction approach. A 100‐percent correlation between the C467T and the C1060‐mutations was found. Fifteen of 17 samples showing the T646A mutation (described earlier in one case of Ax) showed a positive correlation with the C771T mutation in a frequently occurring O1varallele. The two exceptions were defined serologically as Ax.Conclusion:Indications have been found of an evolutionary relationship between A1alleles and Aeland A3subgroups as well as between A2alleles and Aendand Aweaksubgroups. Genetic heterogeneity within the Axand Aintsubgroup

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226142.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:Patients with severe combined immunodeficiency (SCID) treated with allogeneic bone marrow transplantation often receive a milder conditioning regimen than patients who undergo transplantation for hematologic malignancy, and they regularly retain circulating white cells of host origin. The origin of circulating red cells following successful bone marrow transplantation to treat SCID is not known.Study Design and Methods:Review of the medical records identified all patients with SCID who underwent ABO‐mismatched bone marrow transplantation at the University of California, San Francisco, between 1982 and 1994. The ABO and Rh phenotype at>6 months after transplantation was determined for all successful transplants by review of the medical record or the taking of a fresh blood sample for analysis. Patient‐conditioning and donor bone marrow‐preparative regimens were reviewed to assess their possible influence on the red cell phenotype after successful bone marrow transplantation.Results:Nine of 35 SCID patients who underwent successful transplantation received marrow from ABO‐mismatched donors. Eight of the nine patients had only host red cells circulating at 6 to 84 months after transplantation, while one patient had only donor red cells circulating at 48 months after transplantation. None of the patients had circulating red cells of both host and donor origin. Conditioning regimens included cyclophosphamide and antithymocyte globulin for all nine patients; only three patients also received total body irradiation. Seven of the nine patients received related‐donor, HLA‐ mismatched bone marrow, and two patients received HLA‐identical bone marrow; eight patients received T‐cell‐depleted bone marrow. The one patient whose red cell phenotype converted to that of the donor received T‐cell‐depleted, haploidentical marrow, and the preparative regimen included chemotherapy and total body irradiation.Conclusion:SCID patients successfully treated with allogeneic bone marrow transplantation typically fail to show circulating red cells of donor phenotype; this finding is in contrast to the universal presence of circulating donor red cells following successful bone marrow transplantation to treat hematologic malignancies and other diseases. The milder conditioning regimens typically given to patients with SCID, along with T‐cell depletion and HLA mismatching, may play a role in this different outcome. It is not known whether the inability to find circulating red cells of donor origin is due to a failure to engraft donor pluripotent stem cells or a failure of engrafted donor stem cells to differentiate alon

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226143.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background: Reticulocytes are important in the phenotyping of transfused patients. Reticulocytes can persist in blood units for the shelf life of the unit.Study Design and Methods:Temperature dependence of reticulocyte persistence was examined in vitro at 4, 24, and 37°C by using thiazole orange staining and flow cytometric analysis. Two‐color flow cytometric analysis was used to evaluate the persistence of donor reticulocytes in transfused patients.Results:Flow cytometric analysis using thiazole orange demonstrated that persistence of reticulocytes in units of stored CPDA‐1 blood was temperature‐dependent. Reticulocytes disappeared over 13 and 6 days at 24°C and 37°C, respectively, but at 4°C the reticulocyte count changed little over 35 days. Two‐color flow cytometric analysis of reticulocyte antigens was used to follow donor reticulocytes in 14 transfusion events in nine different patients. Donor reticulocytes persisted through 24 hours in 75 percent of the patients and were detectable at 48 hours in three patients.Conclusion:This study demonstrates that reticulocytes persist during refrigerated storage; they are detectable in the circulation of most recipients for the first 24 hours after transfusion and in the circulation of a few recipients after 48 hours. These findings may have relevance for separation techniques based on reticulocyte density in samples drawn shortly after transfusion and for evaluation of reticulocyte counts in patients with hematologic ab

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226144.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:A quality control program of white cell (WBC) reduction in red cells at the bedside was implemented, based on postfiltration counting in a Nageotte chamber of the residual WBCs from samples taken from a segment of the transfusion set, after 1‐in‐10 sample dilution with Turks's solution. During a 1‐year quality control program, 5.1 percent of counted units had apparent filtration failures, that is, WBC counts exceeding 5 × 106per unit. The cause(s) for these apparent failures were investigated.Study Design and Methods:In Study 1, residual WBCs from 150 buffy coat‐free red cells filtered through one type of filter at 4°C, 20 to 24°C, or 27°C in 5 to 10 minutes, 50 to 100 minutes, or 100 to 200 minutes were counted as described above. In Study 2, residual WBCs in samples collected from segments of the transfusion set and from the postfiltration bags were counted in parallel by a new, more sensitive counting method. In this method, 5 mL of filtered red cells was diluted with 20 mL of 3‐percent paraformaldehyde and centrifuged, the pellet was resuspended to 500 μL with a lysis solution, and the WBCs were counted in a Nageotte chamber. In Study 3, residual WBCs were counted by the 3‐percent paraformaldehyde method in samples from postfiltration bags of 1‐ to 2‐ day‐old buffy coat‐rich red cell units filtered through a second type of filter. Filtration was started within 30 minutes of the removal of the unit from the refrigerator, ambient temperature was 20 to 24°C, and the median filtration time was 90 minutes per unit.results:Study 1: Median WBC counts per unit increased progressively from 51,000 at 4°C to 934,000 at 27°C, with intermediate values at 20 to 24°C. In no unit did the WBC count exceed 5 × 106if filtration at 20 to 24°C was completed within 100 minutes, while counts in excess of 50 × 106were found at 20 to 24°C and at 27°C with filtration times of 100 to 200 minutes, and 50 to 100 minutes, respectively. Study 2: The relation between segment and postfiltration bag WBC counts obtained by the 3‐ percent paraformaldehyde method was poor, with the latter being almost always lower than the former. Study 3: None of the 120 units filtered through the second type of filter at 20 to 24°C in 50 to 100 minutes contained more than 3.2 × 106WBCs; the median value was 147,000 WBCs per unit.Conclusion:On the basis of the results with the 3‐percent paraformaldehyde method, which showed the unreliability of segment counts, a new policy was adopted for quality control of bedside WBC reduction, based on controlling the time of and temperature at transfusion. Bedside WBC reduction in 1‐ to 2‐day‐old red cells performed with the second type of filter at 20 to 24°C in less than 100 minutes per unit allowed the preparation of units that meet the standard of fewer than 5 × 106WBCs in all tested cases. Bedside WBC reduction with the second type of filter and under the

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226145.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:The platelet count increases transiently after treatment with polyclonal anti‐D in about 50 percent of D+ patients with autoimmune thrombocytopenic purpura (AITP). The effect is usually attributed to macrophage Fc‐receptor blockade by antibody‐coated red cells. As polyclonal anti‐D is in limited supply, prospective testing was performed on a monoclonal anti‐D (MoAb D) in such patients.Study Design and Methods:Seven D+ patients with chronic AITP received MoAb D intravenously at doses of 47 to 95 μg per kg of body weight. Response was assessed by studying platelet count increment. Hemolysis and red cell‐bound MoAb D were measured before and after MoAb D administration.Results:MoAb D red cell binding was demonstrated in all patients at a ratio higher than that observed in AITP patients successfully treated with polyclonal anti‐D. However, little or no platelet count increment was observed in six patients, while a transient response was observed in only one (platelet count 97 × 109/L before MoAb D infusion and 163 × 109/L 4 days later). Furthermore, because five patients showed signs of hemolysis and two became anemic, higher doses of MoAb D should be used only with caution in patients with AITP.Conclusion:The MoAb D used in this study cannot be proposed as an alternative treatment for pa

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226146.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:About 5 to 10 percent of Asians have platelets that lack the major membrane glycoprotein (GP) IV (CD36, GPIIIb) that carries the isoantigen Naka. The GPIV‐negative platelet phenotype is extremely rare among whites, but its frequency in persons of African ancestry has not yet been determined. Isoimmunization against GPIV can occur in GPIV‐ negative persons and can lead to platelet transfusion refractoriness. Therefore, the expression of GPIV on platelets from unrelated African Americans was studied.Study Design and Methods:Platelets were obtained from 250 African American and 280 white blood donors. Flow cytometry was used to determine the ability of these platelets to bind a monoclonal antibody that reacted with GPIV. Platelets that failed to react with this probe were tested with other GPIV‐specific monoclonal antibodies and with anti‐Naka, an isoantibody that recognizes an epitope on GPIV.Results:Platelets from 6 of the 250 African American donors (2.4%) lacked GPIV and failed to bind anti‐Naka, whereas platelets from all of the white donors were GPIV positive (p>0.05). No platelet‐reactive antibodies were identified in the serum of the GPIV‐ negative donors.Conclusion:The frequency of the GPIV‐negative platelet phenotype in African Americans is comparable to that in Asians and much greater than that in whites. Studies are needed to determine the frequency with which African Americans become isoimmunized to GPIV by transfusions and the possible contribution of this isoimmunization to platelet transfusion refractoriness in

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226147.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY

摘要:

Background:Platelet concentrates and apheresis platelets must be maintained at a temperature as close as possible to 20 to 24°C during transport. To improve temperature control, ensure component quality, and meet handling and freight carrier needs, a new insulated shipping container system was developed and evaluated.Study Design and Methods:Molded polyurethane‐insulated shipping containers were loaded with different payloads of simulated platelet components, with or without gel‐based temperature stabilizing packs (TSPs). The containers were subjected to constant ambient temperature of 37, 4 or ‐10°C. Payload temperatures were continuously monitored, in situ, for 24 hours.Results:Temperature data are reported as the mean number of hours needed for components to warm or cool by 1 degree C. The temperature of payloads exposed to a constant 37°C ambient temperature increased by 1 degree C in 2.5 to 3.8 hours when no TSPs were included in the shipment and in 6.1 to 6.9 hours when TSPs were used. Exposure to a constant 4°C ambient temperature resulted in a 1 degree C temperature decrease in 1.8 to 3.4 hours without TSPs and in 4.6 to 5.6 hours with TSPs. At a ‐10°C ambient temperature, there was a 1 degree C drop within 1.0 to 1.6 hours without TSPs and within 2.7 to 2.9 hours with TSPs.Conclusion:The container and packing methods described moderate the rate of change in the temperature of platelet components during their exposure to challenging ambient conditions. The use of TSPs substantially improves the performance of the system. In addition, the system meets freight carrier requirements and is easy to use, environmentally friendly,

ISSN:0041-1132    

DOI:10.1046/j.1537-2995.1996.36496226148.x

出版商:Blackwell Science Ltd    

年代:1996

数据来源: WILEY