Abstract
Expression of suicide genes (e.g. herpes simplex virus thymidine kinase, HSV-TK) in T cells is an appealing approach to regulate graft-versus-host disease in adoptive immunotherapy. Here we report the optimization of retroviral infection of canine T cells. Canine T cells were stimulated either with phytohemagglutinin (PHA, 2 μg/ml) for 24-72 hours or with 100 U/ml interleukin-2 for seven days. Stimulated cells were co-cultivated with irradiated virus-producing cells. Transduction efficiencies ranged from 4% to 45% using PG13, a gibbon ape leukemia virus envelope (env) pseudotyped packaging cell line. Infection of cells with GPenvAM12, expressing the amphotropic Moloney routine leukemia virus env, did not yield a satisfactory percentage of transduced cells. Enrichment of transduced cells was performed using immunoselection, and gave a purity of up to 98%. Transfusion of 1 10 6 transduced cells per kilogram body weight showed that transduced cells could convert mixed chimerism to 100% and transfer immunity to a specific antigen. Transduced cells were repeatedly detected in peripheral blood and bone marrow by polymerase chain reaction with primers specific for the HSV-TK gene. We have demonstrated the feasibility of using the canine model to study gene therapy as a preclinical model.
Original language | English |
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Pages (from-to) | 25-33 |
Number of pages | 9 |
Journal | Cytokines, Cellular and Molecular Therapy |
Volume | 6 |
Issue number | 1 |
Publication status | Published - 2000 |
Keywords
- Canine model
- Donor leukocyte transfusion
- Gene therapy
- Hematopoietic stem cell transplantation
- Suicide gene (e.g. HSV-TK)
ASJC Scopus subject areas
- Pharmacology
- Immunology and Allergy
- Immunology