L cell and WEHI-3 cell lines will be used for the large scale production of several types of colony stimulating factors. L cell CSF will be purified by a rapid affinity chromatography technique whereas multiple chromotographic and density gradient steps will be utilized for purification of the granulocytic stimulator in WEHI-3 conditioned medium. Antibodies will be prepared against both factors and used to devise radioimmunoassays. The antibodies will be purified by affinity chromatographic techniques. These will be coupled to immunoadsorbents and the resultant gels will be used to concentrate CSF responsive cells. Binding of 125I CSF to marrow cells will be characterized using both macrophages (M) and granulocytic (G) varieties of CSF. Selective use of the purified regulator and cell populations will be utilized to determine whether the progenitor cells are unipotential or have the capability of differentiating into both granulocytes and macrophages. Prostaglandins and isoferritins, putative regulators of cell production, will be tested for possible interactions with CSF receptors. Both GCSF and MCSF will be injected into mice to determine whether serum activity controls the rate of cell production in vivo. Studies will be conducted with diffusion chambers in normal mice and in those with tumor induced granulocytosis to determine whether increased local CSF production may be the primary determinant of granulopoiesis and monocytopoiesis in vivo. Several human cell lines will be established for the large scale production of human-type colony stimulating factor in serum-free medium. These may ultimately be used for the purification of human-active CSF's.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA015237-12
Application #
3164135
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1978-09-01
Project End
1988-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
12
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Montefiore Medical Center (Bronx, NY)
Department
Type
DUNS #
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Shadduck, R K; Fischer, B C; DePasquale, D K et al. (1996) Paradoxical stimulation of normal and leukemic rat hematopoiesis by monoclonal antibody to CSF-1 receptor. Exp Hematol 24:314-7
Logan, T F; Gooding, W; Kirkwood, J M et al. (1996) Tumor necrosis factor administration is associated with increased endogenous production of M-CSF and G-CSF but not GM-CSF in human cancer patients. Exp Hematol 24:49-53
Gilmore, G L; Shadduck, R K (1995) Inhibition of day-12 spleen colony-forming units by a monoclonal antibody to the murine macrophage/monocyte colony-stimulating factor receptor. Blood 85:2731-4
Jones, C M; Poddar, S; Goldstein, R et al. (1994) Human MCF activates monocytes to produce IL-1 but not TNF or CSF-1. Immunobiology 190:303-16
Shadduck, R K; Waheed, A; Mangan, K F et al. (1993) Preparation of a monoclonal antibody directed against the receptor for murine colony-stimulating factor-1. Exp Hematol 21:515-20
Greenberger, J S; Sakakeeny, M A; Leif, J et al. (1992) Expression of M-CSF and its receptor (C-FMS) during factor-independent cell line evolution from hematopoietic progenitor cells cocultivated with gamma irradiated marrow stromal cell lines. Leukemia 6:626-33
Greenberger, J; Leif, J; Crawford, D et al. (1992) Humoral and cell surface interactions during gamma-irradiation leukemogenesis in vitro. Exp Hematol 20:92-102
Shadduck, R K (1992) Granulocyte-macrophage colony-stimulating factor: present use and future directions. Semin Hematol 29:38-42
Shadduck, R K (1992) Granulocyte-macrophage colony-stimulating factor: present status. Semin Hematol 29:1-3
Gonzales-Chambers, R; Przepiorka, D; Shadduck, R K et al. (1991) Autologous bone marrow transplantation with 4-hydroperoxycyclophosphamide-purged marrow for acute lymphoblastic leukemia. Med Pediatr Oncol 19:160-4

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