ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.1

年代:1992

数据来源: MAL

摘要:

In vivotherapeutic manipulation as well asex vivografting are important strategies for bone marrow manipulation and transplantation. This review article discusses the early techniques for marrow processing and manipulation as a background for discussing later graft engineering protocols, as well as preclinical and experimental protocols designed to modify transplantation biology at the multidisciplinary level. The direction of future trials in graft transplantation is also discussed.

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.3

年代:1992

数据来源: MAL

摘要:

High levels of peripheral blood stem cells (PBSC) can be collected from patients with hematopoietic malignancies or solid tumors after high-dose chemotherapy and/or growth factors. Rapid, complete, and durable hematopoietic reconstitution has been observed in patients who have been transplanted with PBSC after conditioning with supralethal myeloablative therapy. We will review the recent data concerning the immunophenotype and functional capacities of mobilized blood stem cells. PBSC seem to be comprised of a mixture of primitive and committed hematopoietic progenitors and are not that different from bone marrow-derived stem cells.

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.19

年代:1992

数据来源: MAL

摘要:

Peripheral blood stem cells (PBSC) can be used for hematopoietic reconstitution in patients who have received high-dose chemotherapy (HDC). Previously, we reported 18 such patients who had nonmobilized PBSC harvested in steady state. We have now performed PBSC reinfusion in 24 patients with hypocellular or tumor-involved marrow who received cyclophosphamide (Cy) mobilization (4 grams/m2) prior to PBSC collection. The PBSC were collected upon rebound hematopoietic recovery by continuous-flow leukapheresis and subsequently cryoproserved. A median of 3.17 X 108mononuclear cells/kg (range 1.47-3.95 X 108) were collected. Patients underwent a median of 6 apheresis procedures (range 5-12). To date all 24 patients have undergone PBSC reinfusion after HDC. Fourteen patients received post-reinfusion granulocyte/macrophage colony-stimulating factor. Granulocyte recovery (>500 x 106/ml) has occurred at a median of 13.5 days (range 7-39) and platelet recovery (>50 X 109/ml without transfusion support) has occurred in 20 patients at a median of 33 days (range 7-121 days). Four other patients have platelet counts of greater than 20K and are transfusion independent, but have still not achieved counts of 50K. PBSC collection after Cy mobilization is feasible and effective for hematopoietic reconstitution after HDC and reinfusion. All 24 patients demonstrated trilineage engraftment. Although neutrophil recovery was not significantly hastened (as compared with our nonmobilized or steady-state data), time to platelet engraftment was foreshortened.

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.27

年代:1992

数据来源: MAL

摘要:

During the last decade, immunomagnetic separation of cells has become popular due to the simplicity and speed of the technology. A variety of magnetic materials have been developed that have fundamentally different characteristics. Some of these act as true colloids and have the properties of solutions, while others are larger and are based on the polymerization of materials, such as styrene, in the presence of magnetite. In this article, the different magnetic matrices are discussed and reviewed with respect to their use for the separation of cells of the hematopoietic system. Additionally, a brief introduction into the field of immunomagnetic separation of nucleic acids is presented, as this technology may be used to perform investigations at the molecular level on hematopoietic cells.

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.35

年代:1992

数据来源: MAL

摘要:

Autologous bone marrow transplantation (ABMT) is the treatment of choice for selected patients with acute myelogenous leukemia, non-Hodgkin's lymphoma, and poor prognosis breast cancer. A possible limitation of this approach is that clonogenic tumor cells could be collected and infused back into the patient along with the normal bone marrow. The major emphasis in our laboratory has been the development of marrow purging regimens for breast cancer patients. This paper describes two investigative approaches hematopoietic progenitor cell protection and selection. We describe how the use of G-CSF in the patients who receive positively selected marrow shortens the rate of engraftment.

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.45

年代:1992

数据来源: MAL

摘要:

The binding of lectins to nucleated cells from human bone marrow was studied in a search for markers that can be used to subdivide further immature hemopoietic cells that are characterized by their expression of CD34. Low-density bone marrow cells were indirectly labeled with biotinylated lectins and streptavidin-R-phycoerythrin (SA-RPE) together with FITC-labeled monoclonal anti-CD34. Four-parameter flow cytometric analysis was then performed and list mode data analyzed. Of the 21 lectins tested, only a few showed differential staining of CD34+versusCD34-cells. These include soybean agglutinin (SBA) andUlex europaeusagglutinin I (UE).Lycopersicon esculentum(LE) andErythrina cristigalli(EC) reacted preferentially with, respectively, CD34+and CD34-cells, suggesting their usefulness in some method to enrich for CD34+cells. This possibility was tested by passing cells labeled with biotinylated lectins over a column containing streptavidin-coated beads. CD34+cells could be enriched>10-fold by competitive (sugar) elution of LE-labeled cells from the column. Similarly, depletion of biotinylated EC-labeled cells by passage through the streptavidin column enriched CD34+cells several fold. The results of these studies document the reactivity of a large panel of lectins with subpopulations of nucleated bone marrow cells and indicate that certain lectins could possibly be used for development of cell separation procedures aimed at the selective enrichment of cells that express CD34.

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.55

年代:1992

数据来源: MAL

摘要:

We have developed a multiassay system consisting of fluorescence microscopy, immunocytology and tumor colony assay to monitor the removal of tumor cells from bone marrow. This system was tested in preclinical purging experiments in which neuroblastoma cells were seeded into bovine marrow and purged by treatment with monoclonal antibodies and immunomagnetic beads. Eight experiments were performed on two different neuroblastoma cell lines seeded at 2% and/or 5% contamination. We consistently demonstrated greater than a 3 log removal with one cycle of antibody/bead treatment and greater than a 1 log further reduction by addition of a second cycle. We also demonstrated removal of all detectable tumor stem cells by this purging method. We feel that this system will prove valuable for monitoringex vivotumor removal in future clinical studies and should be considered for use in other purging trials.

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.65

年代:1992

数据来源: MAL

摘要:

Graft-versus-host-disease (GVHD), which results after allogeneic bone marrow transplantation (BMT), is associated with reduced leukemic relapse. This may be mediated by an immunologic attack with subsequent destruction of residual tumor cells. On the other hand, GVHD does not normally occur after autologous BMT (ABMT), which has an inherently high relapse rate. However, an autoimmune syndrome (AIS) similar to GVHD can be induced after autologous/syngeneic BMT by administration of cyclosporine-A (CsA), resulting in the production of major histocompatibility complex (MHC) class II or Ia autoreactive cytolytic effector cells. Since many hematopoietic malignancies express variable levels of class II molecules, we hypothesized that the adjuvant use of interferon-γ (IFN-γ) with CsA-induced autoimmunity after autologous/syngeneic BMT may upregulate class II antigens on residual tumor cells and make them more susceptible to attack by the la-reactive cells of CsA-induced AIS. The present studies demonstrated that the CsA-induced autoimmune syndrome mediated an anti-tumor effect, although this effect was dependent on challenge with a minimal number of tumor cells. Further studies clearly demonstrated that the antitumor effect could be markedly enhanced by administration of IFN-γ which increased the susceptibility of the tumor to recognition and lysis by the CsA induced autoimmune effector cells. The induction of MHC class II-restricted AIS similar to GVHD by administration of CsA together with the ability to manipulate the surface phenotype of residual tumor cells may lead to decreased relapse rates in the ABMT setti

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.75

年代:1992

数据来源: MAL

摘要:

The high-affinity receptor for IgG, FcγRI, expressed on monocytes and interferon-γ (IFN-γ)-stimulated neutrophils, is a trigger molecule for cell-mediated cytotoxicity. We have prepared murine monoclonal antibodies (MoAb 22 and MoAb 32) that bind to FcγRI outside the ligand binding site and thus bind to and trigger cytotoxicity that is not competed by other immunoglobulins. Because of these properties, it seemed that these MoAbs would be very useful for the development of bispecific antibodies (BsAb) for targeting normal cellular immune defense mechanisms as a new form of immunotherapy for treatment of cancer. BsAbs incorporate into a single molecule the binding specificities of two different antibodies, and, thus, can be used to target myeloid cells to tumors, ensure activation of cellular cytotoxic mechanisms, and target cell lysis and/or phagocytosis. BsAbs were prepared using anti-FeγRI MoAb and an anti-myeloid cell MoAb, PM81, reactive with the CD15 antigen, for studies of antibody-dependent cellular cytotoxicity. Conjugates were made by cross-linking sulfhydryl groups of Fab fragments of MoAb 32 or 22 (both IgG1) and sulfhydryl groups added to intact PM81 (an IgM) usingN-succinimdyl-acetyl-S-thioacetate (SATA). The resulting product was purified by high-performance size-exclusion chromatography. The ability of the BsAbs to mediate attachment of human monocytes to tumor target cells was confirmed in a microtiter well assay of binding of MTT-labeled U937 cells (a human FcγRI-bearing cell line) to SKBR-3 (PM81-reactive breast carcinoma) target cells. The ability of the BsAbs to mediate killing of HL-60 promyelocytic leukemia cells was studied using a 6-hour Chromium-51 release assay. Effector cells were monocytes obtained by cytopheresis and cultured for 18 hours with IFN-γ. Monocytes alone caused minimal killing (5-20%), monocytes plus BsAb caused moderate killing (20-50%), and monocytes plus BsAb plus human serum resulted in maximal killing (50-80%). Experiments were performed to test the ability of the BsAb to purge bone marrow of small numbers of leukemia cells using bone marrow mononuclear phagocytes treated for 18 hours with IFN-γ prior to adding target cells. Without the addition of human serum as a source of complement, a 90% depletion of clonogenic HL-60 cells could be demonstrated. With human complement, up to 95 % depletion was seen. Thus, this BsAb possessed the ability to lyse tumor cell targets by two different mechanisms, complement and cell-mediated lysis. In a Phase I clinical trial, 4 patients with CD15+tumors were treated with up to 48 mg of this BsAb with no toxicity. Of this group, the patient with acute myelogenous leukemia experienced a transient 30-60% reduction in circulating leukemic blast cells during each of six infusions over a 2-week period. Althoughin vitroassays indicated maximal effectiveness between 1 and 10 μg/ml of this BsAb,in vivoreduction in circulating cell counts was observed when peak serum concentrations were as low as 50 ng/ml. As such, this BsAb may be useful forin vivotherapy of high-risk tumors, especially after induction of remission or after bone marrow transplant,i.e., for treatment of minimal residual disease in patients with CD15-expressing tumors, including acute myeloid leukemia, small cell carcinoma of the lung, and colon and breast c

ISSN:1061-6128    

DOI:10.1089/scd.1.1992.1.85

年代:1992

数据来源: MAL