Thymidine kinase (TK) gene-transduced human lymphocytes can be highly purified, remain fully functional, and are killed efficiently with ganciclovir
| Autor: | N C, Munshi, R, Govindarajan, R, Drake, L M, Ding, R, Iyer, R, Saylors, J, Kornbluth, S, Marcus, Y, Chiang, D, Ennist, L, Kwak, C, Reynolds, G, Tricot, B, Barlogie |
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| Rok vydání: | 1997 |
| Předmět: |
Mice
Inbred BALB C Genetic Vectors Cell Separation Flow Cytometry Transfection Antiviral Agents Immunotherapy Adoptive Polymerase Chain Reaction Thymidine Kinase Recombinant Proteins Mice Neoplastic Stem Cells Tumor Cells Cultured Animals Humans Simplexvirus Lymphocytes Enzyme Inhibitors Moloney murine leukemia virus Multiple Myeloma Ganciclovir Cells Cultured Neoplasm Transplantation |
| Zdroj: | Blood. 89(4) |
| ISSN: | 0006-4971 |
| Popis: | A graft-versus-leukemia (GVL) effect has been considered a major factor responsible for cures in patients with hematologic malignancies undergoing allogeneic bone marrow transplantation; however, associated graft-versus-host disease (GVHD) results in significant morbidity and mortality. T-cell depletion reduces the incidence and severity of GVHD but eliminates, at least partially, the GVL effect. Reinfusion of donor T lymphocytes at relapse posttransplantation can induce a potent antitumor response, but GVHD still occurs in the majority of patients. Prior transduction of T lymphocytes with the suicide gene, the viral thymidine kinase (TK), permits specific cell kill on administration of ganciclovir (GCV). Therefore, infusion of TK-transduced T lymphocytes may induce GVL effect and allow for their subsequent selective elimination in case GVHD develops. To evaluate the efficacy and feasibility of this promising approach, anti-CD3-stimulated primary human lymphocytes cultured in interleukin-2 were TK-transduced by a retroviral vector carrying both TK and neomycin-resistance genes. After selection in G418, more than 90% of the cells contained the TK gene as shown by a semiquantitative polymerase chain reaction. In addition, 1 to 5 days of GCV exposure, at clinically achievable concentrations of 20 to 50 micromol/L, inducedor = 90% killing of G418-selected cells without affecting nontransduced cells. Correlation of the extent of T-cell kill and the proportion of TK-gene-transduced cells is consistent with the absence of a bystander effect. Transduced cells were CD3+ and either CD8+ or CD4+ and retained functional properties of untransduced cells. In vivo administration of GCV prevented tumor development after subcutaneous injection of TK-transduced murine myeloma cells (MOPC-11), whereas such an effect was not observed on injection of untransduced cells into the opposite flank. Our studies provide critical information that (1) adequate numbers of TK-transduced lymphocytes can be selected efficiently withor = 90% purity, (2) selected cells remain functional, (3) 24 hours of exposure to GCV at clinically achievable concentration effectsor = 90% killing of selected cells, and (4) GCV is effective in vivo in killing TK-transduced cells. Based on these data, a clinical study has been initiated in patients with multiple myeloma with persistent or relapsing disease after T-cell-depleted allogeneic transplants. |
| Databáze: | OpenAIRE |
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