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

Transplantation of suicide gene modified allogeneic T lymphocytes is an app roach to prevent T cell mediated GVHD while preserving the 'graft-versus-le ukemia' (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 developmen t of more efficient gene transfer protocols into these cells is of great im portance. 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 retrovir al infection. The presented protocol allowed us to obtain transduction rate s of more than 70% of CD3(+) cells after two cycles of infection. It is bas ed on the use of FBS-free media for both the production of retrovirus-conta ining supernatant, as well as the cultivation of the primary T cells. Since the protocol presented here works just as efficiently under large scale co nditions, it may easily he adapted to clinical needs and 'good manufacturin g practice' (GMP) standards.