Retroviral transduction of T lymphocytes for suicide gene therapy in allogeneic stem cell transplantation.
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Retroviral transduction of T lymphocytes for suicide gene therapy in allogeneic stem cell transplantation. / Kühlcke, K; Ayuketang Ayuk, Francis; Li, Z; Lindemann, C; Schilz, A; Schade U, M; Fauser, A A; Zander, A R; Eckert, H G; Fehse, B.
in: BONE MARROW TRANSPL, Jahrgang 25, Nr. 2, 2, 2000, S. 96-98.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Retroviral transduction of T lymphocytes for suicide gene therapy in allogeneic stem cell transplantation.
AU - Kühlcke, K
AU - Ayuketang Ayuk, Francis
AU - Li, Z
AU - Lindemann, C
AU - Schilz, A
AU - Schade U, M
AU - Fauser, A A
AU - Zander, A R
AU - Eckert, H G
AU - Fehse, B
PY - 2000
Y1 - 2000
N2 - 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.
AB - 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.
U2 - 10.1038/sj.bmt.1702364
DO - 10.1038/sj.bmt.1702364
M3 - SCORING: Zeitschriftenaufsatz
VL - 25
SP - 96
EP - 98
JO - BONE MARROW TRANSPL
JF - BONE MARROW TRANSPL
SN - 0268-3369
IS - 2
M1 - 2
ER -
