Efficient gene transfer into spleen cells of newborn mice by a replication-competent retroviral vector

Bachrach, E.; Pelegrin, M.; Piechaczyk, M.; Pedersen, F. S.; Duch, M.


2002-02-15 / vol 293 / pages 328-34


A murine leukemia virus-derived replication-competent retroviral vector with a translational cassette for the enhanced green fluorescence protein (EGFP) was previously found to function efficiently in cell culture (Jespersen et al., 1999, Gene 239, 227-235). We here report that infection of newborn NIH Swiss mice gives rise to EGFP expression in a majority of spleen cells within the first days after infection. Among the nonadherent spleen cells, B, T, and NK lymphocytes were found to be efficiently marked by EGFP by flow cytometry analysis, whereas the adherent spleen cells were negative in most animals. Analysis at time points up to 60 days after infection reveals a decline in EGFP-positive spleen cells over time. Viremia analysis and PCR analysis of spleen DNA indicate that viruses that have lost the translation cassette predominate at later stages. The results provide a model for efficient gene delivery to spleen cells in a time window of 1 to 2 weeks after infection of newborns. Although this type of vector will not in itself be applicable in the clinic, we envision that efficient in vivo gene delivery will be valuable by analysis of differentiation and proliferation of hematopoietic cells in animal models and by development and evaluation of effectors interfering with these processes.

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Animals; Mice; Flow Cytometry; Time Factors; Animals, Newborn; Gene Expression; Leukemia Virus, Murine/*genetics; Gene Transfer Techniques; Green Fluorescent Proteins; Cell Count; *Genetic Vectors; Luminescent Proteins/analysis/genetics; Lymphocytes/metabolism; Spleen/immunology/*metabolism

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