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

Abstract

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 × 10⁶ 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. Bone Marrow Transplantation (2000) 25, Suppl. 2, S96–S98.