Efficient gene transfer into primary murine lymphocytes obviating the need
for drug selection
ML Kuo, N Sutkowski, Y Ron and JP Dougherty
Department of Molecular Genetics and Microbiology, Robert Wood Johnson
Medical School, University of Medicine and Dentistry, Piscataway, NJ
08854-5635.
The efficient introduction of exogenous genes into primary lymphocytes is
potentially important both for somatic cell gene therapy and for studying
lymphocyte biology. We describe the use of retroviral vectors to
efficiently introduce exogenous genes into primary, mature murine lymph
node T and B cells, and primary, immature murine CD4- CD8- double- negative
(DN) thymocytes. Efficient infection of primary cells was achieved by
cocultivation of target cells with lethally irradiated helper cells that
produce high titers of retroviral vectors containing either the neomycin
phosphotransferase II (neo) gene, or both the neo and the human adenosine
deaminase (ADA) genes, in the presence of lymphokines and/or mitogens. Two
days postinfection, without neomycin selection, one to five copies of the
exogenous genes per cell were detected by Southern blot analysis.
Expression of the exogenous human ADA protein was detected at levels
comparable to the endogenous murine ADA protein in the mature T and B
lymphocytes, and was somewhat lower for the immature DN thymocytes.
Volume 82,
Issue 3,
pp. 845-852,
08/01/1993
Copyright © 1993 by The American Society of Hematology
