ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.1

年代:1994

数据来源: MAL

摘要:

We have developed a rapid and accurate method to enumerate the number of CD34+cells in peripheral blood, bone marrow, and leukopheresis samples. The method consists of a two-tube assay and a dedicated software program for data acquisition and analysis. The first reagent combination consists of (a) a nucleic acid dye to identify nucleated cells, (b) a CD45 monoclonal antibody labeled with PE/CY5 to discriminate progenitor cells from mature lymphoid, neutrophil, erythroid, and monocytic cells, (c) an IgG1control antibody labeled with PE to establish the boundary between specific and nonspecific staining, and (d) a known number of fluorescent beads to determine an absolute count of cells. In the second reagent combination the IgG1control antibody is replaced by a CD34 antibody labeled with PE that is used to identify the CD34+cells in the location established by the control reagent combination. The software program uses the fluorescent beads to adjust the forward light scatter, orthogonal light scatter, and three fluorescence detectors of the flow cytometer. The expected location of the CD34+cells is then established with the control reagent combination followed by the enumeration of the CD34+cells per microliter of sample with the reagent combination containing the CD34 antibody. This method is sensitive enough to detect CD34+cells in peripheral blood of normal donors and can reliably determine an increase in CD34+cells in the peripheral blood of patients treated with chemotherapy and/or growth factors. The method alleviates some of the difficulties encountered when small numbers of CD34+cells are enumerated. The system allows for more precise evaluations of the grafts used for bone marrow transplantation.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.3

年代:1994

数据来源: MAL

摘要:

CAMPATH-1 (CDw52) antibodies recognize a very small lipid-anchored glycoprotein that is expressed on the surface of human lymphocytes. They are remarkably lytic with human complement. In addition, CAMPATH-1G (rat IgG2b) and CAMPATH-1H (human IgG1) bind to human Fc receptors and are very effective for cell lysisin vivo. CAMPATH-1M (rat IgM) and CAMPATH-1G have been used to control GVHD and graft rejection in bone marrow transplantation by depletion of the T cells of the donor and recipient. Depletion of donor T cells alone gave excellent control of GVHD but up to 20% of the patients transplanted from HLA-matched siblings, and 51% of those transplanted from nonsibling donors, experienced graft failure caused by immunological rejection. Graft rejection could be partly overcome by additional immunosuppression either with CsA or total lymphoid irradiation (TLI). More effective was the use of CAMPATH-1Gin vivoto deplete residual host lymphocytes. Preliminary results from current protocols of antibody depletion give two year actuarial leukemia-free survival as good as or better than similar studies with conventional GVHD prophylaxis, as well as a decreased morbidity from chronic GVHD, although engraftment was delayed by about 5 days. We propose that prophylactic T cell depletion with CAMPATH-1 antibodies is a simple and valid alternative to drug-based immunosuppression that may be particularly applicable to patients with acute leukemia or nonmalignant diseases transplanted from HLA-matched siblings as well as any patients transplanted from unrelated donors. Future developments of antibody-based immunosuppression may allow the extension of marrow transplantation for tolerance induction to organ transplants or in autoimmune diseases.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.15

年代:1994

数据来源: MAL

摘要:

The neomycin resistance gene in a retroviral vector has been used to mark the marrow of patients receiving autologous bone marrow transplantation for neuroblastoma and acute myeloid leukemia. We have shown that the marker gene can be detected in the resurgent malignant cells at the time of relapse, directly demonstrating that tumorigenic cells contaminate "remission" marrow. We have also shown effective gene transfer to normal primitive progenitor cells and demonstrated a long lasting contribution of the infused marrow to hematopoiesis. The gene marking technique is now being used to evaluate the efficacy of purging and the impact of growth factor treatment on long and short term hematopoietic reconstitution.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.33

年代:1994

数据来源: MAL

摘要:

The plant lectin, soybean agglutinin (SBA), has been widely used to separate heterogeneous populations of cells. In the field of bone marrow transplantation, SBA has been used for partial depletion of T cells from bone marrow allografts to reduce graft-vs.-host disease. SBA's high affinity for many different tumor cells has also indicated its use as a tumor purging agent for autologous bone marrow transplants. We have compared two methods of cell separation using either soluble SBA agglutination, or SBA covalently attached to an activated polystyrene surface. The nonbinding SBA-cell populations generated by these two procedures were very similar in terms of cell recovery, light scatter properties, and phenotypic profile. Notably, both SBA-fractions were enriched in cells with the known progenitor markers, CD34, CD33, and HLA-DR, and were relatively depleted of SBA binding cells. In addition, the activity of each SBA-cell population was measuredin vitroin short-term progenitor assays. Here, both SBA-populations were significantly enriched for CFU-GM. When device-separated SBA-cell populations were seeded into long-term bone marrow culture, they produced both increased progenitor activity and cell proliferation compared to unseparated BMMCs. The polystyrene technology described here could reduce or eliminate many of the drawbacks of soluble SBA agglutination, making SBA cell separation a viable and convenient technique for clinical application.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.37

年代:1994

数据来源: MAL

摘要:

Analysis of the results of different methods of post remission therapy in acute myeloid leukemia (AML) has been used to study their impact on outcome, with a particular emphasis on antileukemic effects by contrasting the risk of relapse. We have compared consolidation chemotherapy (CT), autologous bone marrow transplantation (ABMT) with and without subsequent interleukin-2 (IL-2) and allogeneic bone marrow transplantation (BMT). Additionally we have studied the immunological consequences of each of these maneuvers with particular regard to the cellular components having proven or suspected antileukemic properties and the secondary cytokines released by these cellsin vitroandin vivo.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.47

年代:1994

数据来源: MAL

摘要:

The characterization of tumor cells in bone marrow harvested for autologous bone marrow rescue remains a problem due to the limited sensitivities of the techniques available for their analysis. This complicates the use of purging techniques as their usefulness is debated, specifically because it is not known if they can remove all residual tumor cells from either peripheral blood stem cell harvests or bone marrow. Apart from developing more sensitive techniques for tumor detection, one way of increasing our efficiency at identifying rare malignant cells in normal hematopoietic cells is to develop approaches to enrich the population of interest prior to analysis. Here, we describe a laboratory-based system for tumour enrichment employing panels of monoclonal antibodies (MAbs) and an immunomagnetic colloid. In model systems, tumor cells could be enriched approximately 100-fold with high yield and purity. The adaptations of this technology to permit further cell enrichment and the choice of either an immunological technique or a molecular approach to tumor identification are discussed in detail.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.51

年代:1994

数据来源: MAL

摘要:

Thirty-one marrows were harvested from 29 patients and donors in the first year after startup of a six-bed pediatric bone marrow transplant unit at Children's Memorial Hospital in Chicago. Twenty-seven patients were infused, 13 with allogeneic marrow. Eight of the 14 autologous marrows were purged with 4HC. This paper presents cell processing results from the first year of operation, describes a yield-enhancing change made to the buffy coat procedure, and shows which of various parameters significantly influenced final cell recoveries.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.59

年代:1994

数据来源: MAL

摘要:

The total number and distribution of nucleated cells in harvested bone marrow are potentially important determinants of patient outcome following bone marrow transplantation. In order to assess whether marrows collected from predominantly unrelated donors at Georgetown University Medical Center (GUMC) were different in cellular content from marrows collected at harvest centers outside of GUMC, we compared the nucleated cell counts and mononuclear cell subset distribution (CD34, CD3, CD4, CD8, CD19 antigen-positive cell content) of 10 consecutive marrows harvested at GUMC to 10 unrelated donor marrows from outside harvest centers. Significantly higher nucleated cell counts and CD34 antigen-positive cell content and significantly lower CD3 and CD4 antigen-positive T-cell numbers were demonstrated among the marrows harvested at GUMC. These results confirmed significant variability in marrow collection practices between GUMC and 10 different outside harvest centers and suggest that strict adherence to a specific collection procedure, involving small volume marrow aspirations and multiple puncture sites, results in a product with a high number of early hematopoietic progenitor cells and minimal contamination by peripheral blood. These data further suggest the need for careful monitoring of individual unrelated donor marrow collection centers' practices to optimize the quality of the harvested marrow.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.65

年代:1994

数据来源: MAL

摘要:

Ex vivodepletion of lymphocytes has been used to decrease the incidence of acute graft-versus-host disease following the allogeneic bone marrow transplantation. Many depletion techniques also remove natural killer (NK) cells that may mediate a beneficial graft-versus-leukemia effect. In this paper we review our studies onex vivoexpansion and activation of NK cells for use in the therapy of chronic myelogenous leukemia.

ISSN:1061-6128    

DOI:10.1089/scd.1.1994.3.71

年代:1994

数据来源: MAL