Peripheral blood progenitor cells mobilized by chemotherapy plus granulocyte-colony stimulating factor accelerate both neutrophil and platelet recovery after high-dose VP16, ifosfamide and cisplatin.

We report on the chemotherapy plus granulocyte colony-stimulating factor (G-CSF) induced mobilization of peripheral blood progenitor cells (PBPCs) and their impact on haematopoietic recovery following high-dose chemotherapy. Twenty-four patients with advanced solid tumours or lymphomas received standard-dose chemotherapy with VP16, ifosfamide and cisplatin (VIP) followed by filgrastim (G-CSF; 5 micrograms/kg s.c.

Ex vivo expansion of enriched peripheral blood CD34+ progenitor cells by stem cell factor, interleukin-1 beta (IL-1 beta), IL-6, IL-3, interferon-gamma, and erythropoietin.

To provide sufficient numbers of peripheral blood progenitor cells (PBPCs) for repetitive use after high-dose chemotherapy, we investigated the ability of hematopoietic growth factor combinations to expand the number of clonogenic PBPCs ex vivo. Chemotherapy plus granulocyte colony-stimulating factor (G-CSF) mobilized CD34+ cells from 18 patients with metastatic solid tumors or refractory lymphomas were cultured for up to 28 days in a liquid culture system. The effects of interleukin-1 beta (IL-1), IL-3, IL-6, granulocyte-macrophage-CSF (GM-CSF), G-CSF, macrophage-CSF (M-CSF), stem cell factor (SCF), erythropoietin (EPO), leukemia inhibitory factor (LIF), and interferon-gamma, as well as 36 combinations of these factors were tested.

Haematopoietic growth factors and peripheral blood stem cells as supportive agents in dose intensification.

We have studied the requirements that have to be met to combine effective cancer chemotherapy with the mobilisation of peripheral blood stem cells (PBSC). We have shown that there is a differential induction of high numbers of PBSC following standard-dose chemotherapy (VIP) plus treatment with colony-stimulating factors. The combined sequential administration of interleukin 3 (IL-3) plus granulocyte-macrophage colony-stimulating factor (GM-CSF) induced maximal numbers of PBSC, including colony-forming unit-granulocyte, erythrocyte, monocyte/macrophage, megakaryocyte (CFU-GEMM) and colony-forming unit-megakaryocyte (CFU-Meg), compared with the application of GM-CSF, granulocyte colony-stimulating factor (G-CSF) or chemotherapy alone.

In vivo effects of interleukin-6 on thrombopoiesis in healthy and irradiated primates.

We have studied the in vivo effects of recombinant human interleukin-6 (rhIL-6) on hematopoiesis in eight healthy and nine irradiated cynomolgus monkeys. Of the healthy animals, three received rhIL-6 alone (10 micrograms/kg/d, subcutaneously [SC]), one received rhIL-6 in combination with rhIL-3 (10 micrograms/kg/d, SC), one received rhIL-6 in combination with recombinant cynomolgus granulocyte-macrophage colony-stimulating factor (rcGM-CSF; 10 micrograms/kg/d, SC), two received rhIL-6 in combination with recombinant human granulocyte-CSF (rhG-CSF; 10 micrograms/kg/d, SC), and one received rhIL-6 in combination with recombinant human leukemia inhibitory factor (rhLIF; 10 micrograms/kg/d, SC). All animals were treated for at least 2 weeks with rhIL-6 or the above mentioned combinations.

Sequential administration of interleukin-3 and granulocyte-macrophage colony-stimulating factor following standard-dose combination chemotherapy with etoposide, ifosfamide, and cisplatin.

Purpose: To combine the benefits of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) on neutrophil recovery and recombinant human interleukin-3 (rhIL-3) on platelet recovery, we applied standard-dose chemotherapy with the combined administration of IL-3 and GM-CSF to investigate their efficacy and toxicity.

Patients And Methods: Thirty-six patients with advanced malignancies were treated with etoposide (VP16) 500 mg/m2, ifosfamide 4 g/m2, and cisplatin 50 mg/m2 (VIP), followed by the sequential administration of IL-3 (days 1 to 5 subcutaneously [SC]) and GM-CSF (day 6 to 15 SC). Control patients received GM-CSF alone or were treated without hematopoietic growth factors.

Sequential in vivo treatment with two recombinant human hematopoietic growth factors (interleukin-3 and granulocyte-macrophage colony-stimulating factor) as a new therapeutic modality to stimulate hematopoiesis: results of a phase I study.

In a phase I study, the sequentially administered combination of recombinant human interleukin-3 (rhIL-3) and rhGM-CSF was compared with treatment with rhIL-3 alone in 15 patients with advanced tumors but normal hematopoiesis. Patients were initially treated with rhIL-3 for 15 days. After a treatment-free interval, the patients received a second 5-day cycle of rhIL-3 at an identical dosage, immediately followed by a 10-day course of rhGM-CSF, to assess the toxicity and biologic effects of this sequential rhIL-3/rhGM-CSF combination.

Role of hematopoietic growth factor combinations in experimental and clinical oncology.

Extensive in vitro studies with hematopoietic growth factors as well as numerous clinical trials with single growth factor administration provided the basis for in vivo studies with those factors in combination. Animal models and first clinical trials in humans, with the sequential application of interleukin-3 plus granulocyte-macrophage colony-stimulating factor, demonstrate that growth factor treatment in combination might be effective and could optimize hematologic responses according to specific clinical requirements. This is a brief review of some of the possible clinical applications of hemopoietic growth factors in clinical oncology with particular focus on preclinical and clinical data using combined administration.

Mobilization of peripheral blood progenitor cells by sequential administration of interleukin-3 and granulocyte-macrophage colony-stimulating factor following polychemotherapy with etoposide, ifosfamide, and cisplatin.

We report on the requirements that have to be met to combine a standard-dose chemotherapy regimen with broad antitumor activity with the mobilization of peripheral blood hematopoietic progenitor cells. Thirty-two cancer patients were given a 1-day course of chemotherapy consisting of etoposide (VP16), ifosfamide, and cisplatin (VIP; n = 46 cycles), followed by the combined sequential administration of recombinant human interleukin-3 (rhIL-3) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF). Control patients received GM-CSF alone or were treated without cytokines.

Interleukin-3 and granulocyte-macrophage colony-stimulating factor in combination: clinical implication.

The multilineage hematopoietic effects of IL-3 appear to be most important for its clinical use comprising especially leucocyte and platelet responses. This was demonstrated to be dose dependent characterising doses of 250 to 500 micrograms/m2/day subcutaneously as hematopoietic effective and well tolerable. Since preclinical data suggest synergism between IL-3 and GM-CSF hematopoietic effects of their sequential administration were evaluated in 15 patients with preserved hematopoietic function.

Combination of cytokines: current status and future prospects.

Clinical trials with individual cytokines and extensive in vitro studies have provided the basis for the in vivo use of these molecules in combination. Animal models, with haemopoietic growth factors as well as preliminary studies in humans--as shown by our studies with the sequential use of IL-3 and GM-CSF in patients receiving intensive chemotherapy--indicate that the selection of the appropriate cytokines could optimize haematological responses according to particular clinical requirements. That immunotherapy with IL-2 can induce regression of disseminated human malignancies serves as an encouraging starting point for combinations with other cytokines with the goal of improving the therapeutic efficacy and reducing toxicity.

Fibrinogen gamma-chain mRNA is not detected in human megakaryocytes.

Human megakaryocytes and platelets contain counterparts of several plasma proteins. The origin of most of these alpha-granule proteins is unclear. Fibrinogen represents one of those molecules, being essential in hemostasis, thrombosis, and platelet aggregation.

The role of cytokines in oncology.

The availability of sufficient quantities of recombinant human cytokines and promising preclinical data have led to their introduction into clinical trials. Cytokines have potential as new therapeutic agents in a variety of hematological disorders as well as in solid tumors. Only a few of the still increasing number of these glycoprotein hormones have been studied in humans so far, either as single agents or in combination with chemotherapy and other cytokines.

Hemopoietins in clinical oncology.

Despite major advances in supportive care, neutropenic infections and thrombopenic bleedings remain major lethal treatment- and disease-related complications in patients with malignancy. Moreover, complications of platelet (Plt) and erythrocyte transfusion therapy have become a cause of great concern and shortages of homologous blood products are a constant problem. Suggestions that the application of recombinant human hemopoietins may provide an alternative treatment modality in this patient population is currently being evaluated in clinical trials.

Analysis of megakaryocyte ploidy in patients with thrombocytosis.

The DNA content of bone marrow megakaryocytes was analyzed in 24 patients with myeloproliferative disorders, 23 patients with secondary thrombocytosis and 15 normal volunteers using 2-color flow cytometry. Compared with normal controls, the majority of patients with secondary thrombocytosis, polycythemia vera and essential thrombocytosis exhibited a relative increase in higher ploidy (greater than 16N) cells. In contrast, patients with chronic myelogenous leukemia exhibited an increase in lower ploidy cells (less than 16N), with a modal DNA content of 8N.

Restoration of nonclonal hematopoiesis in chronic myelogenous leukemia with interferon alpha.

Studies have shown that recombinant human alpha interferon (rIFN alpha) inhibits the growth of colonies of multipotential stem cells from human bone marrow. This report demonstrates that rIFN alpha inhibits the growth of such colonies from the bone marrow of patients with chronic myelogenous leukemia (CML) to a greater extent than from bone marrow of healthy individuals. It also shows that T lymphocyte colonies subcloned with interleukin 2 (IL-2) from CML mixed colonies were inhibited more by rIFN alpha than were similar colonies subcultured from normal mixed colonies.

[Proliferation and differentiation in megakaryopoiesis].

Megakaryocytic progenitor cells only complete a limited number of mitotic events; early in megakaryopoiesis, these cells loose their capacity for cell division and acquire the ability for endoreduplication--a phenomenon that is unique to the megakaryocytic lineage. There is growing evidence, that at least two humoral activities affect proliferation of progenitor cells and maturation of its progeny. However, the cytokines that mediate these functionally defined, overlapping activities are not yet precisely known.

Analysis of human hemopoietic progenitor cells for the expression of glycoprotein IIIa.

Human hemopoietic progenitor cells were examined for the expression of glycoprotein IIIa (GPIIIa). This protein, which forms the beta-subunit of the GPIIb/IIIa receptor for cytoadhesive proteins as well as the beta-subunit of the vitronectin receptor, represents the most sensitive cell surface marker so far identified for the megakaryocytic lineage. Bone marrow cells were fractionated by a discontinuous Percoll gradient to separate cells that form megakaryocytic colonies in culture (1.

Megakaryocytic colony-stimulating activity in patients receiving a marrow transplant during hematopoietic reconstitution.

Megakaryocytic colony formation is dependent upon growth-stimulating activities present in human serum or plasma. Factors with diverse biological activities including megakaryocytic colony-stimulating activity (Mk-CSA) are provided by the medium of mitogen stimulated peripheral mononuclear cells or subsets of peripheral T cells. In this communication we describe the stimulatory activity of plasma collected from allogeneic and autologous bone marrow transplant recipients on the growth of megakaryocytic colonies.

Detection of messenger RNAs within single hemopoietic cells by in situ hybridization on small slide areas.

In situ hybridization provides a powerful tool to detect specific mRNA sequences at the cellular level. We have applied a modified in situ hybridization technique using specifically prepared regular glass microscope slides to evaluate mRNA levels in cells of small samples. Cells were derived from in vitro colonies or isolated by fluorescence-activated cell sorting and deposited on the slides.

T-cell depletion with ricin A-chain T101 in allogeneic bone marrow transplantation to prevent severe graft-versus-host disease.

Bone marrow cells from 10 marrow transplant donors were treated with an immunotoxin, which couples A-chain of ricin with a monoclonal anti-T-cell antibody T101 to prevent graft-versus-host disease by the elimination of mature T-cells. Marrow cells treated with the anti human T-cell immunotoxin (IT101) were cultured for erythropoietic colonies, granulocytic colonies, and multilineage hematopoietic colonies (CFU-GEMMT) containing myeloid cells and T-cells, and optimal conditions were defined for the elimination of T-cells present in the harvested donor marrow prior to marrow transplantation. Marrow samples purged with IT101 were examined for residual T-cells by fluorescence activated cell sorting, using anti-T-cell antibodies, [3H]-thymidine incorporation after PHA stimulation, and an assay for clonogenic T-cells.