Long-term expression of gene introduction into normal human T-lymphocytes by retroviral-mediated gene transfer.
Human T-lymphocytes are long lived, easily accessible, mature, and capable of proliferation. They are theoretically a suitable target for retroviral mediated gene transfer. To test this hypothesis, normal human T-cells obtained from bone marrow and peripheral blood were stimulated with phytohemagglutinin (PHA) and infected 24 h later with the retroviral vector N2 which carries the bacterial neo gene. T-lymphocytes were propagated in culture for up to 14 weeks with interleukin-2 (IL-2). Analysis by whole cell RNA dot/blot using a single stranded RNA probe demonstrated persistent expression of the neo gene. Preliminary functional studies revealed that both helper and suppressor functions were preserved in the infected cells in culture. These results demonstrate that normal T-cells are capable of long-term expression of genes introduced by retroviral mediated gene transfer and are potential target cells for somatic gene therapy.
Fauser
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Department of Hematology and Oncology, Albert-Ludwigs-Universität, Freiburg FRG.
D015152:Blotting, Northern; D002455:Cell Division; D002478:Cells, Cultured; D015870:Gene Expression; D005798:Genes, Bacterial; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D016130:Immunophenotyping; D012190:Retroviridae / Q000235:genetics*; D013601:T-Lymphocytes / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection