The growth and proliferation of normal murine peritoneal exudate macrophages (PEM) in vitro is strictly dependent on a macrophage-specific growth factor known as colony-stimulating factor (CSF-1). By contrast, transformed macrophage tumor cell lines do not require exogenous CSF-1 for growth in culture. Recent studies show that tyrosine phosphorylation of certain membrane-bound proteins may be involved in the control of cell proliferation induced by specific growth factors. In addition, several oncogene products have been shown to be structurally related to either specific growth factors or growth factor receptors. We propose to use both normal macrophages and established macrophage cell lines with different growth phenotypes as models to investigate the role of protein phosphorylation in the regulation of macrophage proliferation. First, we will develop two dimensional gel electrophoresis and autoradiography techniques to identify the CSF-1 induced and/or spontaneous phosphorylated components from both mormal macrophages and established macrophage-monocyte tumor cell lines. Thin layer plate electrophoresis techniques will be used to characterize the chemical nature of the phosphorylated amino acids. Second, a monoclonal antibody against a phosphotyrosine moiety will be developed as a probe, along with """"""""Western blot"""""""" techniques, to facilitate the identification and characterization of tyrosine-phosphorylated-proteins. We will study whether the production of autocrine CSF-1 or CSF-like activity and tyrosine phosphorylation are linked to oncogenic transformation of macrophages during the early phase of development. We will establish, by tumor virus transformation, immortalized mouse bone marrow-derived macrophages to study whether these cell lines produce and secrete autostimulatory activity. We will also examine and identify phosphorylated components from these cell lines. The hypothesis of our work is that autocrine growth factor, tyrosine phosphorylation and ligand-receptor interaction play important roles in the cellular transformation and oncogenesis of bone marrow-derived hematopoietic cells.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Wayne State University
Schools of Medicine
United States
Zip Code
Li, Y; Chen, B (1997) Induction of macrophage colony-stimulating factor receptor up-regulation in THP-1 human leukemia cells is dependent on the activation of c-fyn protein tyrosine kinase. Leuk Res 21:539-47
Li, Y; Valeriote, F; Chen, B (1996) Regulation of granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors in a GM-CSF-dependent human myeloid leukemia cell line (AML-193) by interleukin-6. Exp Hematol 24:94-100
Li, Y; Shen, B F; Karanes, C et al. (1995) Association between Lyn protein tyrosine kinase (p53/56lyn) and the beta subunit of the granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors in a GM-CSF-dependent human megakaryocytic leukemia cell line (M-07e). J Immunol 155:2165-74
Li, Y; Chen, B (1995) Differential regulation of fyn-associated protein tyrosine kinase activity by macrophage colony-stimulating factor (M-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF). J Leukoc Biol 57:484-90
Chen, B; Scheding, S; Nakeff, A et al. (1994) Differential expression of mast cell growth factor receptor (c-kit) by peritoneal connective tissue-type mast cells and tissue culture-derived mast cells. J Leukoc Biol 55:596-602
Wu, K F; Rao, Q; Zheng, G G et al. (1994) Enhancement of J6-1 human leukemic cell proliferation by cell-cell contact: role of an M-CSF-like membrane-associated growth factor MAF-J6-1. Leuk Res 18:843-9
Chen, B D; Chou, T H; Sensenbrenner, L (1993) Downregulation of M-CSF receptors by lipopolysaccharide in murine peritoneal exudate macrophages is mediated through a phospholipase C dependent pathway. Exp Hematol 21:623-8
Chen, B; Chou, T H; Sensenbrenner, L (1993) Induction of murine peritoneal macrophage colony-forming cells by peritoneal administration of macrophage inflammatory protein-1 alpha. Exp Hematol 21:1591-6
Fan, K; Barendsen, N; Sensenbrenner, L et al. (1993) Deregulation of granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor in murine macrophage cell line J774A.1. J Cell Physiol 154:535-42
Fan, K; Ruan, Q; Sensenbrenner, L et al. (1992) Transforming growth factor-beta 1 bifunctionally regulates murine macrophage proliferation. Blood 79:1679-85

Showing the most recent 10 out of 24 publications