Establishment of an optimised gene transfer protocol for human primary T lymphocytes according to clinical requirements.
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Establishment of an optimised gene transfer protocol for human primary T lymphocytes according to clinical requirements. / Ayuketang Ayuk, Francis; Li, Z; Kühlcke, K; Lindemann, C; Schade, U; Eckert, H G; Zander, A; Fehse, B.
in: GENE THER, Jahrgang 6, Nr. 10, 10, 1999, S. 1788-1792.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Establishment of an optimised gene transfer protocol for human primary T lymphocytes according to clinical requirements.
AU - Ayuketang Ayuk, Francis
AU - Li, Z
AU - Kühlcke, K
AU - Lindemann, C
AU - Schade, U
AU - Eckert, H G
AU - Zander, A
AU - Fehse, B
PY - 1999
Y1 - 1999
N2 - Current gene therapeutic protocols directed towards the treatment of inherited disorders (eg ADA-SCID) and viral infections (eg AIDS), as well as adoptive immunotherapy approaches are based on the use of genetically modified lymphocytes. Since only insufficient transduction of T cells is obtained using existing techniques, 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 106/ml) by retroviral infection. Using retroviral vectors encoding a truncated human low-affinity nerve growth factor receptor (DeltaLNGFR) as a gene transfer marker, we obtained transduction frequencies of more than 70% of CD3+ cells after two cycles of infection. Our protocol is based on the use of FBS-free media for both the production of retrovirus-containing supernatant and the cultivation of the primary T cells. Since the protocol presented here works just as efficiently under large-scale conditions, it may be easily adapted to clinical needs and 'good manufacturing practice' (GMP) standards.
AB - Current gene therapeutic protocols directed towards the treatment of inherited disorders (eg ADA-SCID) and viral infections (eg AIDS), as well as adoptive immunotherapy approaches are based on the use of genetically modified lymphocytes. Since only insufficient transduction of T cells is obtained using existing techniques, 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 106/ml) by retroviral infection. Using retroviral vectors encoding a truncated human low-affinity nerve growth factor receptor (DeltaLNGFR) as a gene transfer marker, we obtained transduction frequencies of more than 70% of CD3+ cells after two cycles of infection. Our protocol is based on the use of FBS-free media for both the production of retrovirus-containing supernatant and the cultivation of the primary T cells. Since the protocol presented here works just as efficiently under large-scale conditions, it may be easily adapted to clinical needs and 'good manufacturing practice' (GMP) standards.
M3 - SCORING: Zeitschriftenaufsatz
VL - 6
SP - 1788
EP - 1792
JO - GENE THER
JF - GENE THER
SN - 0969-7128
IS - 10
M1 - 10
ER -
