title: Recombinant retrovirus-mediated gene transfer to normal haemopoietic cells.
abstract: Several of parameters seeking to increase the infection frequency of normal haemopoietic cells by recombinant retroviruses containing a drug selectable marker and/or a haemopoietic growth factor gene have been compared. By using bone marrow cells pretreated with 5-fluorouracil, cocultivation of cells for at least 4 days with virus producing fibroblasts, and addition of pokeweed mitogen stimulated spleen cell conditioned medium, from 30 to 100% of haemopoietic spleen colony forming cells (CFU-S) could be infected. Because drug (G-418) selection was found to select subsets of CFU-S, a retrovirus lacking the drug selectable marker Neor, but containing a GM-CSF gene, was used to study the effects of endogenous GM-CSF production on CFU-S differentiation. Infected CFU-S produced equivalent numbers of erythroid and neutrophil-macrophage committed progenitor cells to control uninfected CFU-S. However, the progeny of infected CFU-S produced GM-CSF, some cells grew autonomously and neutrophil numbers were greatly increased at the expense of erythroblasts. These results suggest that unregulated GM-CSF production, although not altering commitment, alters the amplication phase following commitment.
authors.affiliation: Cancer Research Unit, Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
authors.forename: G R
authors.initials: GR
authors.lastname: Johnson
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D003115:Colony-Stimulating Factors / Q000235:genetics* / Q000378:metabolism; D004352:Drug Resistance, Microbial; D005814:Genes, Viral; D005839:Gentamicins / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000235:genetics* / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D051379:Mice; D008815:Mice, Inbred Strains; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection
chemical_list: D000900:Anti-Bacterial Agents; D003115:Colony-Stimulating Factors; D005839:Gentamicins; D006133:Growth Substances; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C010680:antibiotic G 418