We are continuing our efforts to characterize and purify mediators elaborated by human cell lines, e.g., GCT. Our efforts in the current year are aimed at large scale immunization of animals with purified human GM-CSF preparations, possibly using acrylamide gels as added immunogens or alternative coupling methods. We have used several techniques to scale up the purification sequence while minimizing losses. Currently, we have begun to explore the use of hydrophobic HPLC separations and are comparing overall yields between reverse phase supports C4 and C8 and the new phenyl-TSK columns, which may provide similar resolution but avoid the use of solvents such as acetonitrile. Our CSF preparations of highest purity still are able to stimulate erythroid-enhancing activity and mixed Eo-CSF and GM-CSF. We are also investigating factors such as cell concentration, cell cycle, and chemicals such as dimethylsulfoxide on the responsiveness of the leukemia cell-line, HL-60, to CSF. We believe that with careful control of such parameters this population can be used to perform CSF assays using tritiated thymidine incorporation and cytochemical staining techniques, for instance. The availability of such tools will assist us in selecting suitable target cell conditions for study of CSF binding to its receptors, as well as screening for relevant antibodies, polyclonal or monoclonal. (M)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA025512-06
Application #
3166889
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1979-07-01
Project End
1987-10-31
Budget Start
1985-01-01
Budget End
1985-12-31
Support Year
6
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Rochester
Department
Type
Schools of Medicine
DUNS #
208469486
City
Rochester
State
NY
Country
United States
Zip Code
14627
Erickson-Miller, C L; Abboud, C N; Brennan, J K (1988) Purification of GCT cell-derived human colony-stimulating factors. Exp Hematol 16:184-9
Liesveld, J L; Abboud, C N; Slovick, F T et al. (1988) Effect of hydrocortisone and interleukins 1 and 2 on eosinophil progenitors in hypereosinophilic states. Int J Cell Cloning 6:404-16
Liesveld, J L; Abboud, C N; Looney, R J et al. (1988) Expression of IgG Fc receptors in myeloid leukemic cell lines. Effect of colony-stimulating factors and cytokines. J Immunol 140:1527-33
Erickson-Miller, C L; Abboud, C N; Stach, R W et al. (1988) Macrophage colony-stimulating factor in nerve growth factor preparations. J Neurosci Res 19:52-6
Brennan, J K; Lee, K S; Abboud, C N et al. (1987) Interactions of dimethyl sulfoxide and granulocyte-macrophage colony-stimulating factors on the growth and maturation of HL-60 cells. J Cell Physiol 132:246-54
Abboud, C N; Duerst, R E; Frantz, C N et al. (1986) Lysis of human fibroblast colony-forming cells and endothelial cells by monoclonal antibody (6-19) and complement. Blood 68:1196-200
Slovick, F T; Abboud, C N; Brennan, J K et al. (1985) Modulation of in vitro eosinophil progenitors by hydrocortisone: role of accessory cells and interleukins. Blood 66:1072-9
Boros, L; Markham, R E; Brennan, J K (1985) Primary thrombocythemia and acute leukemia. JAMA 253:1721-2
Bloom, R E; Brennan, J K; Sullivan, J L et al. (1985) Lymphoma of host origin in a marrow transplant recipient in remission of acute myeloid leukemia and receiving cyclosporin A. Am J Hematol 18:73-83