Stimulation of G-CSF gene expression in the macrophage cell line by contact with extracellular matrix proteins and a pre-B leukaemia cell line.
In order to scrutinize the adherence-dependent interactions for induction of granulocyte colony-stimulating factor (G-CSF) in peripheral monocytes/macrophages, a sensitive reporter gene assay was constructed using the mouse macrophage cell line transfected with the mouse G-CSF promoter region in conjunction with the luciferase gene as a reporter. With this system, lipopolysaccharide (LPS) showed a markedly positive response. Among the extracellular matrix (ECM) proteins, both fibronectin (FN) and vitronectin (VN) markedly induced luciferase activity, but others did so but much lesser extent. Among the synthetic peptides having Arg-Gly-Asp (RGD) sequences, only FLEPP with multiple RGD significantly induced luciferase activity. Pretreatment of the cells with anti-integrin alpha 6, alpha M, beta 1 and beta 2 monoclonal antibodies (mAbs) significantly reduced the LPS-induced responses and anti-alpha 1, alpha 2 and beta 3 mAbs to lesser extent, and anti-alpha 5, alpha 6, alpha M, beta 1 and beta 2 mAbs blocked the FN-induced response. In the cell-to-cell interactions, significantly positive increase was observed by direct contacting this cell line with a G-CSF-dependent promyelocytic leukaemia cell line, known to stimulate the induction of G-CSF to the stromal cells. Its effect was mostly blocked by pretreatment with anti-integrin alpha 5, alpha L, beta 1 and beta 2 and anti-ICAM-1 mAbs. These results indicate that there are several pathways via the cell-to-ECM and cell-to-cell interactions triggering the induction of G-CSF in the macrophages.
Laboratory of Life Science & Biomolecular Engineering, Japan Tobacco, Inc., Kanagawa, Japan. aoki_y@tcrl.jti.co.jp
Y
Y
Aoki
D
D
Hirano
H
H
Kodama
Y
Y
Nishi
M
M
Nakamura
D000818:Animals; D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D016326:Extracellular Matrix Proteins / Q000378:metabolism* / Q000494:pharmacology; D005353:Fibronectins / Q000378:metabolism / Q000494:pharmacology; D005786:Gene Expression Regulation; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D016023:Integrins / Q000378:metabolism; D015448:Leukemia, B-Cell; D015473:Leukemia, Promyelocytic, Acute; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D009842:Oligopeptides / Q000378:metabolism / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D019096:Vitronectin / Q000378:metabolism / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D016207:Cytokines; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D016023:Integrins; D008070:Lipopolysaccharides; D008934:Mitogens; D009842:Oligopeptides; D019096:Vitronectin; D016179:Granulocyte Colony-Stimulating Factor; C047981:arginyl-glycyl-aspartic acid