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Biomedical subjects

J McCullough

Publications and source records attributed to J McCullough.

At least 217 records · Page 12Linked to original sources

Resolution of red cell compatibility testing problems in patients receiving anti-lymphoblast or anti-thymocyte globulin.

Passively acquired hemagglutinating antibodies may be detected in the serum of patients receiving horse anti-lymphoblast globulin (ALG) or anti-thymocyte globulin (ATG). Thirty-seven patients receiving ALG following renal transplantation were studied. Eight patients monitored daily all developed positive direct antiglobulin tests (DAT) and positive red cell antibody screening tests. Fifteen of 32 recipients developed red cell antibodies reactive at room temperature in saline, 4 of 32 in albumin at 37 degrees C, and 33 of 37 in the antiglobulin test. Horse globulin was detected on the red cells of all six recipients tested with rabbit anti-horse globulin. Ether eluates prepared from the red cells of 20 patients showed no specificity for common red cell antigens. Anti-human globulin (AHG), absorbed with ALG-coated red cells to remove the component in the AHG which was crossreacting with horse globulin, was used successfully for antibody screening and identification, direct antiglobulin testing, and/or the antiglobulin crossmatching of 27 ALG and ATG recipients, including five with red cell alloantibodies.

Animals↗

The frequency of granulocyte-specific antibodies in postpartum sera and a family study of the 6B antigen.

We examined sera from 2313 postpartum women as a potential source of granulocyte specific antibodies. Lymphocyte cytotoxic (LC) antibodies were detected in 397 (17.2%) specimens and granulocyte agglutinating (GA) antibodies in 291 (12.6%). Only two GA positive sera had reactivity for previously defined granulocyte specific antigens (one NA1 and NB1 ). One additional serum had GA reactivity similar to a serum previously reported by van Rood and called anti-6B. The frequency of granulocyte specific antibodies (0.1 percent) in our study indicates that the productivity of random screening of parous sera for granulocyte specific antibodies is low. A four-generation family study of the 6B antigen demonstrated a parallel association between HLA-B7 and B40 and 6B reactivity in the LC, GA, and granulocyte immunofluorescence (GIF) assay. This illustrates that the granulocyte reactivity of anti-6B is not granulocyte specific but is HLA related. This report provides additional evidence that HLA antibodies such as anti-B7 (6B) can react with granulocytes bearing these antigens in the GA and GIF assays currently used to identify granulocyte specific antibodies.

Antibody Specificity↗

Platelet viability following storage for 5 days. Influence of holding whole blood for 8 hours at 20 to 24 degrees C before concentrate preparation.

Regional blood centers frequently need to hold units of whole blood at 20 to 24 degrees C for several hours after phlebotomy so that sufficient platelet concentrates can be prepared to meet the increasing need. We have evaluated the in vivo viability and in vitro properties of platelets that were prepared from whole blood drawn into citrate-phosphate-dextrose-adenine (CPDA-1) either immediately after phlebotomy or after an 8-hour hold at 20 to 24 degrees C. Platelet concentrates were stored for 5 days at 20 to 24 degrees C in polyolefin containers (PL 732, Fenwal) with end-over-end tumbler agitation. The autologous in vivo recovery (mean +/- SD) and one-half disappearance of 51Cr-labeled platelets prepared immediately after phlebotomy were 44.4 +/- 9.4 percent and 4.0 +/- 0.5 days, respectively. Platelets prepared after the delay of 8 hours showed a recovery of 44.5 +/- 8.4 percent and a one-half disappearance of 4.1 +/- 0.4 days. After 5 days of storage, platelet concentrates showed a mean pH of 7.21 +/- 0.20 when prepared immediately after phlebotomy, and of 7.22 +/- 0.15 when prepared after an 8-hour delay. Mean morphology scores were 280 +/- 33 and 302 +/- 27 for platelets from units prepared immediately after phlebotomy or after a holding period of 8 hours, respectively. Platelets underwent synergistic aggregation after 5 days of storage, independent of the length of time that the units of whole blood were held prior to centrifugation. These studies indicate that platelet concentrates prepared from units of whole blood held initially for 8 hours can be stored for 5 days at 20 to 24 degrees C and survive satisfactorily in vivo and retain in vitro characteristics.

Blood Platelets↗

Use of blood components containing red cells by donors of allogeneic bone marrow.

Bone marrow transplantation is performed for an increasing number of disorders. Although much has been written about blood transfusion in bone marrow recipients, there is little information dealing with blood transfusion in bone marrow donors. We examined the records of 129 consecutive allogeneic bone marrow transplants to determine blood use by bone marrow donors. We attempted to correlate blood transfusion requirements with a number of variables in both donors and recipients of bone marrow. The volume of marrow harvested correlated closely with the number of units of blood transfused (r = 0.82). When these two variables were expressed as a percent of estimated blood volume, the correlation improved slightly (r = 0.85). Although the volume of marrow correlated highly with the total number of mononuclear cells harvested (r = 0.92), the correlation between total cells harvested and number of units of blood transfused was not as high (r = 0.75). By knowing the number of marrow cells the recipient requires for transplantation, the volume of donor marrow required and donor blood transfusion requirements can be estimated. In most instances, autologous blood could be used exclusively, thus eliminating the risks associated with homologous blood transfusion.

Adolescent↗

Plasma separation using a hollow fiber membrane device.

A disposable hollow fiber device was evaluated by collecting approximately 550 ml of normal donor plasma (n = 43) and by performing sham (n = 10) and therapeutic (n = 12) plasma exchanges. Blood was processed at 70 ml per min, and plasma flux averaged 23 (collection) and 25 (exchange) ml per min (mean separation efficiencies of 52 and 60%, respectively). The procedures were tolerated well by all donors and patients. The plasma hemoglobin concentration in separated plasma averaged 1 mg per dl, and cell contamination was negligible (mean of 1, 3, and 6 RBCs, platelets and WBCs/microliter, respectively). There was no evidence of in vivo classical or alternative pathway complement activation as assessed by total hemolytic complement generation (CH50), alternative pathway hemolytic activity (AP50), C3 conversion, or C5 activation, nor were unexpected changes seen in the results of laboratory tests performed after the procedure. Sieving coefficients during sham plasma exchange averaged as follows: albumin, 1.03; IgM, 1.0, IgG, 1.0; IgA, 0.98; factor V, 1.07; factor VII, 0.89; factor VIII, 1.05; and factor IX, 1.19. The device appears to be useful for separation of cell-free plasma from blood during therapeutic plasma exchange procedures.

Blood Component Removal↗

Development and operation of a program to obtain volunteer bone marrow donors unrelated to the patient.

Because of recent increases in bone marrow transplantation, the lack of available matched sibling donors has become a problem. This study investigated the feasibility of obtaining volunteer bone marrow donors unrelated to the patient. A program was developed to inform potential donors and obtain their consent for listing in a bone marrow donor registry. Sixty-six percent of whole blood or apheresis donors agreed to participate in the program. There were no demographic differences between those who accepted or declined to participate. Religion, experiences with medical practices, and the attitude of the potential donor's spouse were important factors in the decision to participate. A search for donors was initiated for 127 patients. Thirty searches were not completed, primarily because of patient-related problems. A total of 713 HLA-A and -B antigen-identical donors was located for 63 of the 97 (65%) patients. A total of 40 HLA-A, -B, and -DR antigen-identical donors was located for 18 of these 63 patients. Six of these 18 patients then had one or more donors (total, 9) whose lymphocytes did not react in mixed lymphocyte culture. Thus, using the file of 2147 donors, an HLA-identical donor was located for 6 percent of the 97 patients. None of these patients has yet received a transplant; in four cases this was due to patient-related and in two cases donor-related reasons. The cost to establish the donor program averaged $33.46 (1984) per donor for donors already typed for HLA-A and -B antigens. This experience indicated that, given thorough information about bone marrow donation and transplantation, a majority of blood and apheresis donors will volunteer to donate bone marrow, without feeling pressure to do so. A relatively small donor file provided donors for 6 percent of patients. The long-term value of bone marrow transplantation using unrelated donors and the optimum size of the donor file remain to be established.

Adult↗

Liquid storage of unseparated human bone marrow. Evaluation of hematopoietic progenitors by clonal assay.

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 degrees 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 degrees C, but fell off rapidly after 1 day at 37 degrees C. Recovery of progenitor cells stored 7 days was relatively poor. Best maintenance was achieved for 3 days using 4 degrees C storage without agitation.

Adult↗

Granulocyte transfusion kinetics measured by chemiluminescence, nitroblue tetrazolium reduction, and recovery of indium-111-labeled granulocytes.

A 20-year-old man with chronic granulomatous disease (CGD) and who was receiving granulocyte transfusions for a refractory liver abscess was studied to compare the kinetics of 111In-labeled granulocytes with those of two functional granulocyte assays, nitroblue tetrazolium reduction and chemiluminescence. Transfused granulocytes were eliminated in both rapid and slow phases. Peak recovery was noted in the first sample, which was obtained 10 minutes after transfusion for each assay. The elimination kinetics were similar over 24 hours. These results confirm the value of using 111In-labeled granulocytes as a marker of transfused granulocytes. These data also confirm that the oxidative metabolic function of granulocytes prepared by continuous-flow leukapheresis remains intact while in the recipient's circulation. The response of the patient adds support for the use of granulocyte transfusions in certain patients with CGD.

Adult↗

Three-hour collection of committed and multipotent hematopoietic progenitor cells by apheresis.

Although multipotent progenitor cells have been collected from blood in humans by apheresis in 1- to 2-hour collections, longer-term collections of multipotent progenitor cells have not been reported. To determine whether such collections could be sustained, the authors performed six 3-hour collections on five normal volunteers, processing a mean of 8024 ml of blood. Blood samples were taken at 0, 1.5, and 3 hours, and component samples were taken at 1.5 and 3 hours. Cell counts, differentials, and committed myeloid (CFU-GM), erythroid (BFU-E), and multipotent (CFU-MIX or CFU-GEMM) progenitor cell assays were performed. Collection efficiency for progenitor cells, the number of cells collected divided by the number in the blood processed, did not diminish significantly. The number collected in the second 1.5-hour period resembled the number collected during the first. Circulating numbers did not fall significantly during apheresis. Prolongation of apheresis collections may reliably increase the number of progenitor cells available for eventual hematopoietic reconstitution.

Blood Component Removal↗

Effect of ABO incompatibility on the fate in vivo of 111Indium granulocytes.

The effect of ABO incompatibility on the in vivo fate of 111Indium granulocytes was determined. The intravascular recovery and survival (t1/2) and extravascular migration into a skin window of normal-donor granulocytes did not differ in 15 subjects from the values obtained in four controls who received ABO-compatible granulocytes. Nor was the correlation between the ABO antibody titers and the in vivo measurements strongly positive. It is concluded that ABO incompatibility did not alter the in vivo fate of granulocytes.

ABO Blood-Group System↗

HLA class I and II antigen phenotypes of North American Indians from Minnesota: implications for marrow transplants using unrelated donors.

As a result of an appeal for a bone marrow donor for a North American Indian (Native American) patient, 261 Native Americans from our community were typed for HLA-A,B,DR antigens, and 51 were typed for HLA-A,B antigens only. The HLA antigen frequencies of the Native Americans were compared with those of 12,881 white bone marrow donors and were found to differ markedly. To investigate the implications these differences in HLA antigen frequencies would have for the location of unrelated bone marrow donors, the HLA types of 12 Native American bone marrow transplant patients from our institution were used to search among 5389 HLA-A,B,DR-typed white donors in the National Marrow Donor Program file and the file of 261 HLA-A,B,DR-typed Native American donors. In the white donor file, at least two donors were found that matched at all HLA-A,B,DR antigen loci of one Native American patient (8%). Using the Native American donor file, which was less than one-twentieth the size of the white donor file, and HLA-A,B,DR-matched donor was also found for one (8%) of the patients. These results suggest that although donors for nonwhites can be identified in a file of HLA-typed white volunteers, the probability of finding a suitably matched donor for such individuals is enhanced if donors representing racial or ethnic minorities are included in unrelated donor registries.

Bone Marrow Transplantation↗

Adverse reactions in patients transfused with cryopreserved marrow.

Marrow is cryopreserved for use in autologous bone marrow transplants, but little is known of the incidence of reactions in patients transfused with these cryopreserved marrows. Reactions in patients transfused during a 4-year period with 134 autologous marrows cryopreserved in dimethyl sulfoxide (DMSO) were compared with those in patients transfused with marrow that had been collected from HLA-compatible donors and that had not been cryopreserved. Patients transfused with cryopreserved marrow had significantly more nausea (44.8 vs. 14.1%; p less than 0.0005), vomiting (23.9 vs. 8.5%; p less than 0.01), chills (31.3 vs. 1.4%; p less than 0.0005), and fever (17.9 vs. 0%; p less than 0.005) than patients transfused with fresh allogeneic marrow. The incidence of emesis correlated with the dose of DMSO received, but that of nausea did not. All cryopreserved marrows were cultured for bacteria at the time of transfusion and 17 (12.7%) were found to be positive. Only 1 of the 17 patients transfused with culture-positive marrow developed sepsis during the transplant course with the same organism that was present in the transfused marrow. Although the reactions in donors transfused with cryopreserved marrow were readily treated, this study suggests that the incidence of some reactions might be decreased by reducing the dose of DMSO transfused. Bacterial contamination of transfused marrow was a worrisome complication, and efforts should be made to improve marrow collection and processing techniques to minimize that risk.

Bacteria↗

Effects on donors of second bone marrow collections.

Second bone marrow transplants have been used successfully to treat marrow graft failure or relapse following the original marrow transplantation. Hospital records of 16 related two-time bone marrow donors were reviewed to determine what risks a person faces in making a second marrow donation. One donor suffered a minor complication following the first collection, but no donors suffered complications during the second donation. The volume of marrow collected for the first and second donations (1253 +/- 504 vs. 1261 +/- 471 mL) was similar. However, more marrow donors received transfusions of homologous red cells during the second collection than during the first collection (5/16 compared to 1/16; p = 0.06). To determine what factors might contribute to the high incidence of homologous red cell transfusion during second donations, the experiences of donors who received homologous blood during the second donation were compared to the experiences of donors who received only autologous blood. Persons receiving homologous blood during the second collection received more red cell units than persons receiving autologous blood (2.0 +/- 0.7 units compared to 0.9 +/- 0.5 units; p less than 0.01). Their precollection hemoglobin levels were lower (12.3 +/- 1.2 vs. 14.4 +/- 1.4 g/dL [123 +/- 12 vs. 144 +/- 14 g/L]; p less than 0.01), and the time between the first and second collections was significantly less (45 +/- 8 days vs. 342 +/- 432 days; p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Mislabeling of elutriation medium: a demonstration of the need to improve the quality control of reagents produced for cell processing.

BACKGROUND: The processing of hematopoietic cells to remove red cells, T-lymphocytes, or malignant cells has become common. However, few of the reagents used are produced specifically for marrow processing and approved by the Food and Drug Administration. CASE REPORTS: A labeling error is reported in a commercially manufactured, research-grade reagent used in an elutriation procedure to deplete T cells from mononuclear cells obtained from bone marrow for allogeneic transplantation. The elutriation medium used in the procedure had been labeled as containing 0.9-percent sodium chloride, 0.1-percent dextrose, and 0.3 mM EDTA, but it actually contained 3.0 mM EDTA. When this medium was not available from the manufacturer, it was prepared in this hospital, according to the stated formulation, with 0.3 mM EDTA. When marrows from two unrelated donors were processed with this 0.3 mM EDTA-containing medium, cell loss due to aggregation occurred, and, in one recipient, the marrow did not engraft. After the error in the medium composition was discovered, marrow was collected a second time from the same donor. The marrow was processed with medium containing 3.0 mM EDTA, and no cell aggregation was observed. CONCLUSION: Food and Drug Administration-approved reagents, prepared specifically for use in cell processing and produced according to good manufacturing practices, should be used whenever possible.

Bone Marrow Cells↗