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.
CRC Department of Medical Oncology, Beatson Institute for Cancer Research, Glasgow, UK.
W N
WN
Keith
R
R
Brown
I B
IB
Pragnell
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