title: Isolation and properties of gene-modified mouse bcr-abl-transformed cells expressing various immunostimulatory factors.
abstract: B210 cells are murine (BALB/c) cells transformed by bcr-abl fusion gene. After intravenous administration they are capable of inducing leukaemia-like disease in syngeneic mice. From these cells a thymidine-kinase less subline was derived. It was significantly less pathogenic than the parental cells. However, a highly pathogenic clone denoted B210cTK-/cl-2 was isolated from its population. As determined by Western blotting, these cells produced more p210 protein than the parental B210 cells. To successfully transfect these cells a modified electroporation method was introduced. Bicistronic plasmids carrying gene for herpes simplex thymidine kinase (HSV TK) and the gene for either granulocyte-monocyte colony stimulation factor (GM-CSF), interleukin-2 (IL-2) or interleukin 12 (IL-12) were used for the transfection experiments. Gradually, cell lines producing these cytokines were isolated in media supplemented with hypoxantin, aminopterin and thymidine (HAT). All of them were highly sensitive to ganciclovir in vitro confirming that the cells produced HSV TK. The genetic modification of B210cTK-/cl-2 was associated neither with the alteration of p210 bcr-abl production nor with any changes in expression of MHC class I molecules. From populations of each of the three lines several cell clones were isolated and tested for the production of the respective cytokines. The original uncloned population and several clones differing in the cytokine production were administered intravenously into mice. All animals survived without symptoms of the disease suggesting that the gene-modification was associated with the loss of pathogenicity.
  
  
  BACKGROUND
  CML, Bcr-Abl, HSV TK, cytokines, gene-modified tumour cells, pathogenicity.
authors.affiliation: Department of Experimental Virology, Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
authors.forename: M
authors.initials: M
authors.lastname: Petrackova
authors.forename: E
authors.initials: E
authors.lastname: Sobotkova
authors.forename: M
authors.initials: M
authors.lastname: Duskova
authors.forename: P
authors.initials: P
authors.lastname: Jinoch
authors.forename: V
authors.initials: V
authors.lastname: Vonka
mesh_terms: D000276:Adjuvants, Immunologic / Q000235:genetics*; D000818:Animals; D002471:Cell Transformation, Neoplastic; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics*; D016044:Fusion Proteins, bcr-abl / Q000032:analysis; D015774:Ganciclovir / Q000627:therapeutic use; D016313:Genes, abl; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
chemical_list: D000276:Adjuvants, Immunologic; D016207:Cytokines; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D013937:Thymidine Kinase; D016044:Fusion Proteins, bcr-abl; D015774:Ganciclovir