Retrovirus mediated transfer and expression of GM-CSF in haematopoietic cells.
Two retrovirus vectors were compared for their ability to express granulocyte-macrophage colony stimulating factor (GM-CSF) in a haematopoietic cell line, FDCP1, which is dependent on GM-CSF for survival. Both a MoMLV-based vector pVneoGM, and a MPSV-based vector, M3neoGM, were found to be capable of transmitting and expressing both GM-CSF and neomycin sequences in the myeloid FDCP1 cell line. Our results also demonstrate that pVneoGM is more efficient at generating GM-CSF independent colonies than M3neoGM. Analysis of cell lines derived after infection confirmed pVneoGM expressed higher levels of GM-CSF. Cell lines generated by infection with pVneoGM responded to levels of exogenous recombinant GM-CSF which did not stimulate growth of the parental cell line, suggesting autocrine stimulation may convey a proliferative advantage under sub-optimal growth conditions. Finally the parental vectors pVneo and M3neo were shown to be capable of expressing the neomycin gene in both murine haematopoietic progenitor and stem cells.
Keith
W N
WN
CRC Department of Medical Oncology, Beatson Institute for Cancer Research, Glasgow, UK.
Brown
R
R
Pragnell
I B
IB
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D002874:Chromosome Mapping; D004247:DNA / Q000032:analysis; D015870:Gene Expression; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis*; D006133:Growth Substances / Q000096:biosynthesis*; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics*; D012313:RNA / Q000032:analysis; D009053:Sarcoma Viruses, Murine / Q000235:genetics*; D014162:Transfection
D006133:Growth Substances; D012313:RNA; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA