Efficient gene transfer into spleen cells of newborn mice by a replication-competent retroviral vector.
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.
Bachrach
Estanislao
E
Institut de Génétique Moléculaire, UMR 5535/IFR24, CNRS, 1919, Route de Mende, 34293 Montpellier Cedex 05, France.
Pelegrin
Mireia
M
Piechaczyk
Marc
M
Pedersen
Finn Skou
FS
Duch
Mogens
M
D000818:Animals; D000831:Animals, Newborn; D002452:Cell Count; D005434:Flow Cytometry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008214:Lymphocytes / Q000378:metabolism; D051379:Mice; D013154:Spleen / Q000276:immunology / Q000378:metabolism*; D013997:Time Factors
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins