Journal of Leukocyte Biology
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(Journal of Leukocyte Biology. 2000;68:757-764.)
© 2000 by Society for Leukocyte Biology

Selective stimulation of G-CSF gene expression in macrophages by a stimulatory monoclonal antibody as detected by a luciferase reporter gene assay

Yoshiko Aoki, Shiken Sha, Hidehito Mukai and Yoshisuke Nishi

Laboratory of Life Science & Biomolecular Engineering, Japan Tobacco Inc., Yokohama, Kanagawa, Japan

Correspondence: Yoshisuke Nishi, Ph.D., Director, Laboratory of Life Science & Biomolecular Engineering, Japan Tobacco, Inc., 6-2 Umegaoka, Aoba-ku, Yokohama, Kanagawa 227-8512, Japan. E-mail: yoshisuke.nishi{at}ims.jti.co.jp

We have identified a stimulatory monoclonal antibody (mAb) from autoimmune mice that selectively stimulates granulocyte colony-stimulating factor (G-CSF) gene expression in a mouse macrophage cell line. The induction was observed not only in the cell line, but also in normal peritoneal macrophages. This mAb bound to the monocyte/macrophage cell lines and pre-B leukemia cell lines, but also in normal peritoneal macrophages, whereas it did not bind to normal T and B cells in the spleen or fibroblastic cell lines. It could even bind to a human promyelocytic leukemia cell line, when they were differentiated into monocytic cells. On Western blotting, this mAb mainly recognized an approximately 30-kDa band and it was unique because there have been no reports of membrane-associated proteins with a similar molecular mass found in macrophages. These results suggest that there could be a specific gateway molecule to induce G-CSF in macrophages.

Key Words: RAW264.7 • peritoneal exudate macrophages






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