High efficiency gene transfer and expression in normal murine B lymphocytes.
The introduction of new genetic information into hematopoietic cells offers a new approach for investigating the molecular events controlling differentiation. Retrovirus vectors have been used to transfer new genes with high efficiency into murine hematopoietic cells, primarily of the myeloid lineage. In this report, we show that vectors carrying the dominant, selectable gene for neomycin resistance (neo gene) can successfully infect normal murine B lymphocytes (CFU-B). The infected CFU-B formed colonies in vitro in high concentrations (750 micrograms/ml) of G418, a neomycin analogue. That B lymphocytes contained the neo gene was confirmed by the findings that the putative B cell colonies growing in G418 contained antibody-producing cells and that the cells responding to the B cell mitogen, LPS, were resistant to G418. Infection of normal spleen cells with different vectors containing a variety of transcriptional regulatory sequences resulted in 7-40% of the CFU-B becoming G418 resistant. Introduction of the immunoglobulin heavy chain enhancer into NEO vectors appeared to augment the expression of the neo gene, since the level of G418 resistance was higher in B cells infected with a NEO vector containing the enhancer than in cells infected with a vector lacking the enhancer.
Wu
D D
DD
Huszar
D
D
Dick
J E
JE
Bernstein
A
A
Phillips
R A
RA
D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D002454:Cell Differentiation; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D005260:Female; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000276:immunology; D006462:Hemolytic Plaque Technique; D007136:Immunoglobulins / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008808:Mice, Inbred CBA; D009355:Neomycin / Q000494:pharmacology*; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic
D005839:Gentamicins; D007136:Immunoglobulins; D008070:Lipopolysaccharides; C010680:antibiotic G 418; D009355:Neomycin