Background and Objectives. Gene manipulation and cell vaccines represent innovative strategies to enhance the immunogenicity of cancer cells. We adopted a defective lentivirus derived from the human immunodeficiency virus (HIV)-1 backbone and carrying the enhanced green fluorescent protein (EGFP) gene to transduce primary human acute myelogenous leukemia (AML) and B-precursor acute lymphoblastic leukemia (ALL) cells. Design and Methods. AML blasts were maintained with or without cytokines (stem cell factor, FLT3 ligand and interleukin 3) for 48 hours, and successively infected with two spin infection cycles. ALL blasts were cultured on a murine S17 stromal cell line. Results. As regards AML cells, the efficiency of infection at 7 days varied from 8.4 to 37%. As confirmed by cell cycle analysis, cells were, in most of the cases, blocked in different phases of the cycle and did not proliferate during culture: the infection was therefore obtained in the absence of cell proliferation. In c...
Biagi, E., Bambacioni, F., Gaipa, G., Casati, C., Golay, J., Biondi, A., et al. (2001). Efficient lentiviral transduction of primary human acute myelogenous and lymphoblastic leukemia cells. HAEMATOLOGICA, 86(1), 13-16.
Efficient lentiviral transduction of primary human acute myelogenous and lymphoblastic leukemia cells
BIAGI, ETTORE;GAIPA, GIUSEPPE;BIONDI, ANDREA;
2001
Abstract
Background and Objectives. Gene manipulation and cell vaccines represent innovative strategies to enhance the immunogenicity of cancer cells. We adopted a defective lentivirus derived from the human immunodeficiency virus (HIV)-1 backbone and carrying the enhanced green fluorescent protein (EGFP) gene to transduce primary human acute myelogenous leukemia (AML) and B-precursor acute lymphoblastic leukemia (ALL) cells. Design and Methods. AML blasts were maintained with or without cytokines (stem cell factor, FLT3 ligand and interleukin 3) for 48 hours, and successively infected with two spin infection cycles. ALL blasts were cultured on a murine S17 stromal cell line. Results. As regards AML cells, the efficiency of infection at 7 days varied from 8.4 to 37%. As confirmed by cell cycle analysis, cells were, in most of the cases, blocked in different phases of the cycle and did not proliferate during culture: the infection was therefore obtained in the absence of cell proliferation. In c...I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.