21 results on '"Leimig, T."'
Search Results
2. Large-scale isolation of CD133+ progenitor cells from G-CSF mobilized peripheral blood stem cells
- Author
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Gordon, P R, Leimig, T, Babarin-Dorner, A, Houston, J, Holladay, M, Mueller, I, Geiger, T, and Handgretinger, R
- Published
- 2003
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3. A large-scale method for T cell depletion: towards graft engineering of mobilized peripheral blood stem cells
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Gordon, PR, Leimig, T, Mueller, I, Babarin-Dorner, A, Holladay, MA, Houston, J, Kerst, G, Geiger, T, and Handgretinger, R
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- 2002
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4. Predominance and characteristics of Burkitt lymphoma among children with non-Hodgkin lymphoma in northeastern Brazil
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Sandlund, JT, Fonseca, T, Leimig, T, Verissimo, L, Ribeiro, R, Lira, V, Berard, CW, Sixbey, J, Crist, WM, Mao, L, Chen, G, Pui, C-H, Heim, M, and Pedrosa, F
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- 1997
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5. Efficient in vitro generation of adult multipotent cells from mobilized peripheral blood CD133+ cells
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Kuçi, S., Kuçi, Z., Schmid, S., Seitz, G., Müller, I., Dufke, A., Leimig, T., Murti, G., Jurecic, R., Schumm, M., Lang, P., Bruchelt, G., Bader, P., Klingebiel, T., Niethammer, D., and Handgretinger, R.
- Published
- 2008
6. A one-step large-scale method for T- and B-cell depletion of mobilized PBSC for allogeneic transplantation
- Author
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Barfield, R.C., Otto, M., Houston, J., Holladay, M., Geiger, T., Martin, J., Leimig, T., Gordon, P., Chen, X., and Handgretinger, R.
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- 2004
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7. Purification of human natural killer cells using a clinical-scale immunomagnetic method
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Iyengar, R., Handgretinger, R., Babarin-Dorner, A., Leimig, T., Otto, M., Geiger, T.L., Holladay, M.S., Houston, J., and Leung, W.
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- 2003
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8. 803 - Process Development and Manufacturing: NOVEL SOFTWARE FOR ROBUST AND REPRODUCIBLE CLINICAL GRADE ENRICHMENT OF T REGULATORY CELLS EMPLOYING THE CLINIMACS PRODIGY.
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Akel, S., Leimig, T., Porter, V., Hu, X., Peppenelli, M., Paszkiet, B., Gottschalk, S., Triplett, B., and Riberdy, J.M.
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REGULATORY T cells , *MANUFACTURING processes - Published
- 2023
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9. Efficient in vitro generation of adult multipotent cells from mobilized peripheral blood CD133+ cells.
- Author
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Kuçi, S., Kuçi, Z., Schmid, S., Seitz, G., Müller, I., Dufke, A., Leimig, T., Murti, G., Jurecic, R., Schumm, M., Lang, P., Bruchelt, G., Bader, P., Klingebiel, T., Niethammer, D., and Handgretinger, R.
- Subjects
BLOOD cells ,STEM cells ,CELL proliferation ,ASTROCYTES ,CELL cycle ,LIVER cells - Abstract
Objectives: To generate non-haematopoietic tissues from mobilized haematopoietic CD133
+ stem cells. Materials and methods: Mobilized peripheral blood CD133+ cells from adult healthy donors were used. In vitro ability of highly enriched CD133+ cells from mobilized peripheral blood to generate multipotent cells, and their potential to give rise to cells with characteristics of neuroectoderm, endoderm and mesoderm layers was investigated. Results: We found that a recently identified population of CD45+ adherent cells generated in vitro after culture of highly purified CD133+ cells for 3–5 weeks with Flt3/Flk2 ligand and interleukin-6 can, in presence of the appropriate microenvironmental cues, differentiate into neural progenitor-like cells (NPLCs), hepatocyte-like cells and skeletal muscle-like cells. We have termed them to be adult multipotent haematopoietic cells (AMHCs). AMHC-derived NPLCs expressed morphological, phenotypic and molecular markers associated with primary neural progenitor cells. They can differentiate into astrocyte-like cells, neuronal-like cells and oligodendrocyte-like cells. Moreover, AMHC-derived NPLCs produced 3,4-dihydrophenylalanine and dopamine and expressed voltage-activated ion channels, suggesting their functional maturation. In addition, AMHC-derived hepatocyte-like cells and skeletal muscle-like cells, showed typical morphological features and expressed primary tissue-associated proteins. Conclusion: Our data demonstrate that AMHCs may therefore serve as a novel source of adult multipotent cells for autologous replacement cell therapies. [ABSTRACT FROM AUTHOR]- Published
- 2008
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10. Large-scale isolation of CD133+progenitor cells from G-CSF mobilized peripheral blood stem cells.
- Author
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Gordon, P.R., Leimig, T., Babarin-Dorner, A., Houston, J., Holladay, M., Mueller, I., Geiger, T., and Handgretinger, R.
- Subjects
STEM cells ,CELL transplantation ,LEUKAPHERESIS - Abstract
We have evaluated the feasibility of large-scale isolation of CD133+ progenitors from healthy mobilized adult donors for potential clinical use in autologous and allogeneic transplantation. A total of 11 healthy volunteer adult donors were mobilized with G-CSF. CD133 + stem cells were isolated from a single leukapheresis using the Clinimacs method. The median percentage of CD133 before positive selection was 0.75% (range 0.39-2.03%). After selection, the median purity and recovery was 94% (range 85.2-98.0%) and 69% (range 44-100%), respectively. The median log10 T-cell depletion obtained by CD133+ positive selection was 4.2 (range 3.8-4.7). The CD133+ progenitors were highly enriched in colony-forming units (CFU) and transplantation into NOD/SCID mice resulted in a high engraftment rate. Transplantation of sorted CD133 +/CD34 + cells into NOD/SCID mice showed a higher engraftment compared to CD133-/CD34+ cells. Mobilized peripheral CD133+ stem cells can be purified in large scale for potential clinical use. The biological function of the cells is not impaired. The majority of the NOD/SCID repopulating cells are within the CD133 +/CD34 + subpopulation. Therefore, clinical studies using purified CD133 + stem cells can be envisoued to further clarify the role of CD133+ stem cells in hematopoietic reconstitution after transplantation. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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11. New Forms of Transplantation.
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Handgretinger, R., Leimig, T., Babarin-Domer, A., Holladay, M.A., Gordon, J. Houston P., Corbacioglu, S., Greil, J., laws, H., Dilloo, D., Strahm, B., Peters, C., Sykora, K., Luethy, A. Ridolfi, Friedrich, W., Gungor, T., Schulz, A., Wachowiak, J., Chybicka, A., and Boruczkowski, D.
- Subjects
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TRANSPLANTATION of organs, tissues, etc. , *STEM cell transplantation - Abstract
Bone Marrow Transplantation (2002) 30, S11–S15. doi:10.1038/sj.bmt.1703744 [ABSTRACT FROM AUTHOR]
- Published
- 2002
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12. Combination immunotherapy with clinical-scale enriched human gammadelta T cells, hu14.18 antibody, and the immunocytokine Fc-IL7 in disseminated neuroblastoma.
- Author
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Otto M, Barfield RC, Martin WJ, Iyengar R, Leung W, Leimig T, Chaleff S, Gillies SD, and Handgretinger R
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- Animals, Antibodies, Monoclonal immunology, Blood Donors, Cell Survival drug effects, Cell Survival immunology, Cytotoxicity Tests, Immunologic, Female, Flow Cytometry, Gangliosides immunology, Granulocyte Colony-Stimulating Factor pharmacology, Humans, Interleukin-7 immunology, Leukapheresis, Mice, Mice, Inbred NOD, Mice, SCID, Neuroblastoma immunology, Neuroblastoma secondary, T-Lymphocytes, Cytotoxic metabolism, Transplantation, Heterologous, Antibodies, Monoclonal pharmacology, Cytotoxicity, Immunologic immunology, Immunotherapy, Neuroblastoma prevention & control, Receptors, Antigen, T-Cell, gamma-delta immunology, T-Lymphocyte Subsets immunology, T-Lymphocytes, Cytotoxic immunology
- Abstract
Purpose: To evaluate a combined cellular and humoral immunotherapy regimen in a mouse model of disseminated human neuroblastoma. We tested combinations of clinical-grade, isolated human gammadelta T cells with the humanized anti-GD2 antibody hu14.18 and a novel fusion cytokine, Fc-IL7., Experimental Design: gammadelta T cells were large-scale enriched from leukapheresis product obtained from granulocyte colony-stimulating factor-mobilized donors. gammadelta T cell cytotoxicity was tested in a europium-TDA release assay. The effect of Fc-IL7 on gammadelta T-cell survival in vitro was assessed by flow cytometry. NOD.CB17-Prkdc(scid)/J mice received 1 x 10(6) NB-1691 neuroblastoma cells via the tail vein 5 to 6 days before therapy began. Treatment, for five consecutive weeks, consisted of injections of 1 x 10(6) gammadelta T cells weekly, 1 x 10(6) gammadelta T cells weekly, and 20 microg hu14.18 antibody four times per week, or 1 x 10(6) gammadelta T cells weekly with 20 microg hu14.18 antibody four times per week, and 20 mug Fc-IL7 once weekly., Results: The natural cytotoxicity of gammadelta T cells to NB-1691 cells in vitro was dramatically enhanced by hu14.18 antibody. Fc-IL7 effectively kept cultured gammadelta T cells viable. Combination therapy with gammadelta T cells and hu14.18 antibody significantly enhanced survival (P = 0.001), as did treatment with gammadelta T cells, hu14.18 antibody, and Fc-IL7 (P = 0.005). Inclusion of Fc-IL7 offered an additional survival benefit (P=0.04)., Conclusions: We have shown a new and promising immunotherapy regimen for neuroblastoma that requires clinical evaluation. Our approach might also serve as a therapeutic model for other malignancies.
- Published
- 2005
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13. Phenotype and function of human natural killer cells purified by using a clinical-scale immunomagnetic method.
- Author
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Leung W, Iyengar R, Leimig T, Holladay MS, Houston J, and Handgretinger R
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- Animals, Antilymphocyte Serum pharmacology, Cell Adhesion Molecules metabolism, Cell Proliferation, Cytokines metabolism, Graft vs Host Disease immunology, Granulocyte Colony-Stimulating Factor pharmacology, Granzymes, Humans, Immunomagnetic Separation, Membrane Glycoproteins metabolism, Mice, Mice, Inbred NOD, Mice, SCID, Perforin, Pore Forming Cytotoxic Proteins, Receptors, Immunologic immunology, Serine Endopeptidases metabolism, Immunophenotyping, Killer Cells, Natural immunology
- Abstract
Infection, disease relapse, graft failure, and graft-versus-host disease (GVHD) are significant adverse events associated with allogeneic bone marrow transplantation. Donor natural killer (NK) cells may be an ideal cell type for prevention or treatment of all these adverse events. Therefore, we investigated the phenotype and function of human NK cells purified by using a clinical-scale immunomagnetic method. We found that the NK cell purification procedures did not adversely affect the expression of killer cell immunoglobulin-like receptors, adhesion molecules, intracellular cytokines, perforin, and granzyme B. Purified NK cells had extensive proliferative capacity and potent antitumor activity when assessed using an immunodeficient mouse model. While all mice transplanted with unpurified mononuclear cells developed GVHD, none of the mice transplanted with purified NK cells did. NK cells were highly susceptible to lysis by antithymocyte globulin (ATG), whereas G-CSF had a minimal effect on their natural cytotoxicity. These results support future clinical investigation of the use of purified NK cells for adoptive immunotherapy in the absence of ATG.
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- 2005
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14. Biology and plasticity of CD133+ hematopoietic stem cells.
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Handgretinger R, Gordon PR, Leimig T, Chen X, Buhring HJ, Niethammer D, and Kuci S
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- AC133 Antigen, Adult, Animals, Antigens, CD, Antigens, CD34 metabolism, Cell Division, Cell Size, Flow Cytometry, Humans, Mice, Mice, SCID, Transplantation, Heterologous, Cell Differentiation, Glycoproteins metabolism, Hematopoietic Stem Cells cytology, Peptides metabolism
- Abstract
AC133 (CD133) is a highly conserved antigen expressed on hematopoietic stem cells with unknown function. In order to further characterize CD133(+) progenitor cells, we purified CD133(+) stem cells using the method of magnetic activated cell sorting (MACS) from healthy adult volunteers mobilized with granulocyte colony-stimulating growth factor (G-CSF) to a mean purity of 94%. The purified CD133(+) cells highly engrafted NOD/SCID mice. In addition, unseparated mononuclear cells or CD133(+) stem cells isolated from the bone marrow of transplanted NOD/SCID mice gave rise to engraftment of secondary recipients. Upon ex vivo culture of purified CD133(+) cells with FLT3/Flk2 ligand (FL) and interleukin-6 (IL-6), a plastic-adherent cell population could be observed after 6 weeks in culture. These adherent cells did not express CD34 or CD133 antigens on their surface, nor did they express markers for endothelial, mesenchymal, or dendritic cells. After incubation of these adherent cells with stem cell factor (SCF), non-adherent cells were observed which partially co-expressed CD133, but were negative for CD34. These nonadherent CD34(-) cells showed a high engraftment capacity in NOD/SCID mice. From our results, we conclude that CD133 might be a marker of early progenitors with a high NOD/SCID engraftment potential. The fact that CD133(+) hematopoietic progenitors can give rise to an adherent population which is CD133(-) and CD34(-) and that these cells can again give rise to a CD133(+)CD34(-) stem cell population with high NOD/SCID engraftment potential indicates the plasticity of hematopoietic precursors. CD133(+) stem cells might be useful for research and for clinical application.
- Published
- 2003
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15. Functional amelioration of murine galactosialidosis by genetically modified bone marrow hematopoietic progenitor cells.
- Author
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Leimig T, Mann L, Martin Mdel P, Bonten E, Persons D, Knowles J, Allay JA, Cunningham J, Nienhuis AW, Smeyne R, and d'Azzo A
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- Animals, Ataxia etiology, Ataxia therapy, Bone Marrow Cells cytology, Carboxypeptidases administration & dosage, Carboxypeptidases genetics, Carboxypeptidases pharmacokinetics, Cathepsin A, Central Nervous System Diseases etiology, Central Nervous System Diseases therapy, Genetic Therapy methods, Green Fluorescent Proteins, Hematopoietic Stem Cell Transplantation, Kidney Diseases etiology, Kidney Diseases therapy, Luminescent Proteins genetics, Mice, Mice, Knockout, Mucolipidoses complications, Mucolipidoses pathology, Neuraminidase deficiency, Organ Specificity, Tissue Distribution, Treatment Outcome, beta-Galactosidase deficiency, Hematopoietic Stem Cells metabolism, Lysosomal Storage Diseases therapy, Mucolipidoses therapy
- Abstract
Protective protein/cathepsin A (PPCA), a lysosomal carboxypeptidase, is deficient in the neurodegenerative lysosomal disorder galactosialidosis (GS). PPCA(-/-) mice display a disease course similar to that of severe human GS, resulting in nephropathy, ataxia, and premature death. Bone marrow transplantation (BMT) in mutant animals using transgenic BM overexpressing the corrective enzyme in either erythroid cells or monocytes/macrophages has proven effective for the improvement of the phenotype, and encouraged the use of genetically modified BM cells for ex vivo gene therapy of GS. Here, we established stable donor hematopoiesis in PPCA(-/-) mice that received hematopoietic progenitors transduced with a murine stem cell virus (MSCV)-based, bicistronic retroviral vector overexpressing PPCA and the green fluorescent protein (GFP) marker. We observed complete correction of the disease phenotype in the systemic organs up to 10 months after transplantation. PPCA(+) BM-derived cells were detected in all tissues, with the highest expression in liver, spleen, BM, thymus, and lung. In addition, a lysosomal immunostaining was seen in nonhematopoietic cells, indicating efficient uptake of the corrective protein by these cells and cross-correction. Expression in the brain occurred throughout the parenchyma but was mainly localized on perivascular areas. However, PPCA expression in the central nervous system was apparently sufficient to delay the onset of Purkinje cell degeneration and to correct the ataxia. The long-term expression and internalization of the PPCA by cells of systemic organs and the clear improvement of the neurologic phenotype support the use of this approach for the treatment of GS in humans. (Blood. 2002;99:3169-3178)
- Published
- 2002
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16. Retrovirus-mediated gene transfer of the cytokine genes interleukin-1beta and tumor necrosis factor-alpha into human neuroblastoma cells: consequences for cell line behavior and immunomodulatory properties.
- Author
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Coze C, Leimig T, Jimeno MT, and Mannoni P
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- Cell Division, Coculture Techniques, Genetic Vectors, Humans, Lymphocyte Culture Test, Mixed, Lymphocyte Subsets, Neuroblastoma pathology, Phenotype, Tumor Cells, Cultured, Gene Transfer Techniques, Interleukin-1 genetics, Neuroblastoma genetics, Retroviridae genetics, Tumor Necrosis Factor-alpha genetics
- Abstract
We have investigated the value of a gene therapy approach for neuroblastoma (NB), based on retroviral transduction of the IL-1beta or TNF-alpha cytokine genes into human NB lines. Secretion of the corresponding cytokine, was demonstrated in all lines, although with considerable quantitative variations. Cytokine gene expression significantly reduced the proliferation index (p = 0.0001); this effect was associated with either terminal neuronal (one TNF-alpha line) or fibroblast-like differentiation (two IL-1beta lines), leading to growth arrest after a few weeks. Cell surface levels of CD54 and HLA class II remained unaffected, but HLA class I (p < 0.001) and CD58 expression (p = 0.01) increased on SKNSH after TNF-alpha gene transfer. Mononuclear cells from normal allogeneic donors cocultured with both IL-1beta (p < 0.001) and TNF-alpha lines (p < 0.01), showed a significant increase in the proportion of activated T cells (CD3+DR+); however, their cytotoxicity and proliferation rate remained unchanged. Immunotherapy of neuroblastoma will require identification of transduced lines in which cytokine secretion induces phenotypic changes in such a way as to augment their likely immunomodulatory properties without impeding cell growth. Because of the limited efficacy of IL-1beta or TNF-alpha gene transfer alone, further studies should focus on combination with other immunomodulatory agents, to improve their potential efficacy in neuroblastoma.
- Published
- 2001
17. IL-2 adenovector-transduced autologous tumor cells induce antitumor immune responses in patients with neuroblastoma.
- Author
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Bowman L, Grossmann M, Rill D, Brown M, Zhong WY, Alexander B, Leimig T, Coustan-Smith E, Campana D, Jenkins J, Woods D, Kitchingman G, Vanin E, and Brenner M
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- Adolescent, Child, Child, Preschool, Humans, Infant, Interleukin-2 therapeutic use, Killer Cells, Natural immunology, Longitudinal Studies, Neuroblastoma pathology, Neuroblastoma therapy, T-Lymphocyte Subsets immunology, T-Lymphocytes, Cytotoxic immunology, T-Lymphocytes, Helper-Inducer immunology, Transfection immunology, Adenoviridae genetics, Genetic Vectors therapeutic use, Immunotherapy, Adoptive methods, Interleukin-2 genetics, Neuroblastoma genetics, Neuroblastoma immunology
- Abstract
In many different murine models, the immunogenicity of tumor cells can be increased by transduction with a range of immunostimulatory genes, inducing an immune response that causes regression of pre-existing unmodified tumor cells. To investigate the relevance of these animal models to pediatric malignancy, we used autologous unirradiated tumor cells transduced with an adenovirus-IL-2 to immunize 10 children with advanced neuroblastoma. In a dose-escalation study, we found that this tumor immunogen induced a moderate local inflammatory response consisting predominantly of CD4(+) T lymphocytes, and a systemic response, with a rise in circulating CD25(+) and DR+ CD3(+) T cells. Patients also made a specific antitumor response, manifest by an IgG antitumor antibody and increased cytotoxic T-cell killing of autologous tumor cells. Clinically, five patients had tumor responses after the tumor immunogen alone (one complete tumor response, one partial response, and three with stable disease). Four of these five patients were shown to have coexisting antitumor cytotoxic activity, as opposed to only one of the patients with nonresponsive disease. These results show a promising correlation between preclinical observations and clinical outcome in this disease, and support further exploration of the approach for malignant diseases of children., (Copyright 1998 by The American Society of Hematology.)
- Published
- 1998
18. Interleukin-2 gene-modified allogeneic tumor cells for treatment of relapsed neuroblastoma.
- Author
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Bowman LC, Grossmann M, Rill D, Brown M, Zhong WY, Alexander B, Leimig T, Coustan-Smith E, Campana D, Jenkins J, Woods D, and Brenner M
- Subjects
- Cell Transplantation, Child, Child, Preschool, Humans, Injections, Subcutaneous, Neuroblastoma pathology, Phenotype, Recurrence, Transplantation, Homologous, Treatment Outcome, Tumor Cells, Cultured, Cancer Vaccines, Cytotoxicity, Immunologic, Genetic Therapy, Interleukin-2 biosynthesis, Interleukin-2 genetics, Neuroblastoma immunology, Neuroblastoma therapy
- Abstract
Tumor cells that have been genetically modified to express immunostimulatory genes will induce effective antitumor responses in a range of syngeneic animal models. For human applications, transduced autologous tumor cell lines are often difficult or impossible to prepare, so that there are strong incentives for substituting a standardized allogeneic tumor cell line. However, such lines may be inferior immunogens if they differ from host tumors in the antigens they express. We have evaluated the safety, immunostimulatory, and antitumor activity of an interleukin-2-secreting allogeneic neuroblastoma cell line in 12 children with relapsed stage IV neuroblastoma. They received two to four subcutaneous injections of cells in a dose-escalating schedule, up to a maximum of 10(8) cells per injection. There was induration and pruritus at the injection site, and skin biopsies revealed mild panniculitis with CD3+ cells surrounding scanty residual tumor cells. There was a limited but significant peripheral monocytosis. No patient showed any increase in direct cytotoxic effector function against the immunizing cell line, but 3 patients had a rise in the frequency of neuroblastoma-reactive cytotoxic T lymphocyte precursor cells. One child had > 90% tumor response (PR), 7 had stable disease, and 4 had progressive disease in response to vaccine alone. Although these results offer some encouragement for the continued pursuit of allogeneic vaccine strategies in human cancer, the antitumor immune responses we observed are inferior to those obtained in an earlier immunization study using autologous neuroblastoma cells. Hence, we suggest that this earlier approach remains preferable, its difficulties notwithstanding.
- Published
- 1998
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19. Gene marking and gene therapy for transplantation medicine.
- Author
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Dilloo D, Rill DR, Grossmann ME, Leimig T, and Brenner MK
- Subjects
- Biomarkers, Gene Transfer Techniques, Humans, Genetic Therapy, Hematopoietic Stem Cell Transplantation methods
- Abstract
The classic application for gene therapy is in the correction of single gene defects, although this has been complicated by the low efficiency of gene transfer into hematopoietic cells. Gene therapy, however, has potential for the modulation of tumor cell growth, drug sensitivity, and antitumor immune responses. In addition, gene marking can be used, in spite of this limited transfer efficiency, to provide information on hematopoiesis, sources of cancer relapse after stem cell transplant, and the relative efficacy of graft manipulation techniques. This article reviews the applications of gene therapy and gene marking in transplantation medicine.
- Published
- 1996
- Full Text
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20. High-efficiency transduction of freshly isolated human tumor cells using adenoviral interleukin-2 vectors.
- Author
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Leimig T, Brenner M, Ramsey J, Vanin E, Blaese M, and Dilloo D
- Subjects
- Coculture Techniques, Gene Deletion, HLA-DR Antigens analysis, Humans, Interleukin-2 biosynthesis, Lac Operon genetics, Lymphocytes immunology, Retroviridae, T-Lymphocyte Subsets, T-Lymphocytes immunology, T-Lymphocytes, Cytotoxic immunology, Tumor Cells, Cultured, Adenoviruses, Human genetics, Gene Transfer Techniques, Genetic Vectors genetics, Interleukin-2 genetics, Neuroblastoma immunology, Neuroblastoma metabolism
- Abstract
Tumor cells genetically modified to express immunostimulatory molecules can produce high levels of antitumor immunity in rodent models. Although a number of clinical trials are currently in progress to assess the value of the approach in human disease, almost all require ex vivo transduction of cultured tumor cells with retroviral vectors. This process is not feasible for many human malignancies, hampering clinical evaluation of the approach. We have used an E1a,1b/E3 deletion mutant of adenovirus containing either the lacZ or the human interleukin-2 (IL-2) gene to transduce human neuroblastoma cells. This vector transduces fresh neuroblastoma cells and neuroblastoma cell lines with an efficiency of 80-90%, compared to an efficiency of 0-14% obtained with retroviral vectors. Cells transduced with the IL-2 adenovector produce up to 12,000 pg of IL-2/10(6) cells/24 hr. IL-2 adenovector-transduced neuroblasts are immunostimulatory; when they are cultured with patient lymphocytes, they increase the proportion of DR+ T cells and generate major histocompatibility complex (MHC) unrestricted cytotoxic effector cells active against parental (nontransduced) tumor cells. We conclude that IL-2 adenovector can be used to transduce freshly isolated human tumor cells efficiently, which will then produce immunomodulatory quantities of the cytokine. The use of adenoviral rather than retroviral vectors facilitates preparation of human tumor "vaccines" and these vectors are now being used in our clinical study of neuroblastoma patients.
- Published
- 1996
- Full Text
- View/download PDF
21. Immunomodulatory effects of human neuroblastoma cells transduced with a retroviral vector encoding interleukin-2.
- Author
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Leimig T, Foreman N, Rill D, Coze C, Holladay M, and Brenner M
- Subjects
- Antibodies, Monoclonal immunology, Antibodies, Monoclonal pharmacology, Antigens, CD analysis, DNA, Complementary genetics, Genes, Reporter, Humans, Immunotherapy, Interleukin-2 immunology, Interleukin-2 metabolism, Kanamycin Kinase, Lymphocyte Subsets immunology, Neuroblastoma immunology, Phosphotransferases (Alcohol Group Acceptor) biosynthesis, Phosphotransferases (Alcohol Group Acceptor) genetics, Recombinant Fusion Proteins biosynthesis, Transfection, Tumor Cells, Cultured, Vaccination, Genetic Vectors genetics, Interleukin-2 genetics, Neuroblastoma pathology, Retroviridae genetics
- Abstract
We have investigated whether retroviral mediated transfer of the IL-2 gene renders human neuroblastoma cells immunogenic, justifying their use in a clinical tumor immunization study. Fourteen neuroblastoma cell lines were established from patients with disseminated neuroblastoma and transduced with the vector G1Ncvl2, which contains the neomycin phosphotransferase gene and the cDNA of the human interleukin-2 gene. Clones secreting > 150 pg/10(6) cells/24 h of IL-2 were selected for further study. Secretion of IL-2 was maintained for at least 3 weeks in nonselective media, implying that production of the cytokine would continue under in vivo conditions. Co-culture of IL-2 transduced cell lines with patient lymphocytes induced potent cytotoxic activity against both transduced and parental neuroblastoma cell lines. This activity was HLA unrestricted, and predominantly mediated by CD16+ or CD56+ and CD8- lymphocytes. These data form the preclinical justification for our current immunization protocol for patients with relapsed or resistant neuroblastoma.
- Published
- 1994
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