Retroviral transduction of T lymphocytes for suicide gene therapy in allogeneic stem cell transplantation

• Published in: Bone marrow transplantation (Basingstoke) Vol. 25 Suppl 2; no. S2; pp. S96 - S98
• Main Authors: Kühlcke, K, Ayuk, F A, Li, Z, Lindemann, C, Schilz, A, Schade, U M, Fauser, A A, Zander, A R, Eckert, H G, Fehse, B
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01-05-2000
• Subjects: Bone marrow; Bone marrow transplantation; CD3 antigen; Fibronectins; Gene therapy; Genetic Therapy - methods; Good Manufacturing Practice; Graft vs Host Disease - prevention & control; Graft vs Leukemia Effect; Graft-versus-host reaction; Hematopoietic Stem Cell Transplantation; Humans; In Vitro Techniques; Infections; Leukemia; Lymphocytes; Lymphocytes T; Protocol; Retroviridae - genetics; Retrovirus; Stem cell transplantation; Stem cells; Suicide; Suicide genes; T-Lymphocytes - virology; Transduction; Transduction, Genetic; Transplantation; Transplantation, Homologous
• ISSN: 0268-3369; 1476-5365
• DOI: 10.1038/sj.bmt.1702364

Transplantation of suicide gene modified allogeneic T lymphocytes is an approach to prevent T cell mediated GVHD while preserving the 'graft-versus-leukemia' (GVL) effect of an allograft. A prerequisite for such a therapy is the efficient transduction of T cells with suitable vectors. Since existing techniques allow only insufficient transduction of T cells, the development of more efficient gene transfer protocols into these cells is of great importance. We present here a protocol for the highly efficient transduction of human primary T cells at high densities (1 x 10(6) cells/ml) by retroviral infection. The presented protocol allowed us to obtain transduction rates of more than 70% of CD3+ cells after two cycles of infection. It is based on the use of FBS-free media for both the production of retrovirus-containing supernatant, as well as the cultivation of the primary T cells. Since the protocol presented here works just as efficiently under large scale conditions, it may easily be adapted to clinical needs and 'good manufacturing practice' (GMP) standards.