Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Mar;82(5):1526–1530. doi: 10.1073/pnas.82.5.1526

Purification and biochemical characterization of human pluripotent hematopoietic colony-stimulating factor.

K Welte, E Platzer, L Lu, J L Gabrilove, E Levi, R Mertelsmann, M A Moore
PMCID: PMC397296  PMID: 3871951

Abstract

Pluripotent hematopoietic colony-stimulating factor (pluripotent CSF), a protein that is constitutively produced by the human bladder carcinoma cell line 5637, has been purified from low serum (0.2% fetal calf serum)-containing conditioned medium. The purification involved sequential ammonium sulfate precipitation, ion-exchange chromatography, gel filtration, and reversed-phase high-performance liquid chromatography. The purified protein has a molecular weight of 18,000 in NaDodSO4/polyacrylamide gel electrophoresis, both by the silver staining technique and by elution of biological activity from a corresponding gel slice, and has an isoelectric point of 5.5. Pluripotent CSF supports the growth of human mixed colonies, granulocyte-macrophage colonies, and early erythroid colonies and induces differentiation of the human promyelocytic leukemic cell line HL-60 and the murine myelomonocytic leukemic cell line WEHI-3B (D+). The specific activity of the purified pluripotent CSF in the granulocyte-macrophage colony assay is 1.5 X 10(8) units/mg of protein.

Full text

PDF
1526

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abboud C. N., Brennan J. K., Barlow G. H., Lichtman M. A. Hydrophobic adsorption chromatography of colony-stimulating activities and erythroid-enhancing activity from the human monocyte-like cell line, GCT. Blood. 1981 Dec;58(6):1148–1154. [PubMed] [Google Scholar]
  2. Broxmeyer H. E., Moore M. A., Ralph P. Cell-free granulocyte colony inhibiting activity derived from human polymorphonuclear neutrophils. Exp Hematol. 1977 Mar;5(2):87–102. [PubMed] [Google Scholar]
  3. Burgess A. W., Metcalf D. The nature and action of granulocyte-macrophage colony stimulating factors. Blood. 1980 Dec;56(6):947–958. [PubMed] [Google Scholar]
  4. Das S. K., Stanley E. R., Guilbert L. J., Forman L. W. Human colony-stimulating factor (CSF-1) radioimmunoassay: resolution of three subclasses of human colony-stimulating factors. Blood. 1981 Sep;58(3):630–641. [PubMed] [Google Scholar]
  5. Das S. K., Stanley E. R. Structure-function studies of a colony stimulating factor (CSF-1). J Biol Chem. 1982 Nov 25;257(22):13679–13684. [PubMed] [Google Scholar]
  6. Fauser A. A., Messner H. A. Granuloerythropoietic colonies in human bone marrow, peripheral blood, and cord blood. Blood. 1978 Dec;52(6):1243–1248. [PubMed] [Google Scholar]
  7. Fauser A. A., Messner H. A. Identification of megakaryocytes, macrophages, and eosinophils in colonies of human bone marrow containing neurtophilic granulocytes and erythroblasts. Blood. 1979 May;53(5):1023–1027. [PubMed] [Google Scholar]
  8. Fauser A. A., Messner H. A., Lusis A. J., Golde D. W. Stimulatory activity for human pluripotent hemopoietic progenitors produced by a human T-lymphocyte cell line. Stem Cells. 1981;1(2):73–80. [PubMed] [Google Scholar]
  9. Fung M. C., Hapel A. J., Ymer S., Cohen D. R., Johnson R. M., Campbell H. D., Young I. G. Molecular cloning of cDNA for murine interleukin-3. Nature. 1984 Jan 19;307(5948):233–237. doi: 10.1038/307233a0. [DOI] [PubMed] [Google Scholar]
  10. Golde D. W., Bersch N., Quan S. G., Lusis A. J. Production of erythroid-potentiating activity by a human T-lymphoblast cell line. Proc Natl Acad Sci U S A. 1980 Jan;77(1):593–596. doi: 10.1073/pnas.77.1.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gough N. M., Gough J., Metcalf D., Kelso A., Grail D., Nicola N. A., Burgess A. W., Dunn A. R. Molecular cloning of cDNA encoding a murine haematopoietic growth regulator, granulocyte-macrophage colony stimulating factor. 1984 Jun 28-Jul 4Nature. 309(5971):763–767. doi: 10.1038/309763a0. [DOI] [PubMed] [Google Scholar]
  12. Ihle J. N., Keller J., Henderson L., Klein F., Palaszynski E. Procedures for the purification of interleukin 3 to homogeneity. J Immunol. 1982 Dec;129(6):2431–2436. [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lu L., Broxmeyer H. E., Meyers P. A., Moore M. A., Thaler H. T. Association of cell cycle expression of Ia-like antigenic determinations on normal human multipotential (CFU-GEMM) and erythroid (BFU-E) progenitor cells with regulation in vitro by acidic isoferritins. Blood. 1983 Feb;61(2):250–256. [PubMed] [Google Scholar]
  16. Lu L., Broxmeyer H. E. The selective enhancing influence of hemin and products of human erythrocytes on colony formation by human multipotential (CFUGEMM) and erythroid (BFUE) progenitor cells in vitro. Exp Hematol. 1983 Sep;11(8):721–729. [PubMed] [Google Scholar]
  17. Lusis A. J., Quon D. H., Golde D. W. Purification and characterization of a human T-lymphocyte-derived granulocyte-macrophage colony-stimulating factor. Blood. 1981 Jan;57(1):13–21. [PubMed] [Google Scholar]
  18. Messner H. A., Izaquirre C. A., Jamal N. Identification of T lymphocytes in human mixed hemopoietic colonies. Blood. 1981 Aug;58(2):402–405. [PubMed] [Google Scholar]
  19. Metcalf D. Clonal extinction of myelomonocytic leukemic cells by serum from mice injected with endotoxin. Int J Cancer. 1980 Feb 15;25(2):225–233. doi: 10.1002/ijc.2910250210. [DOI] [PubMed] [Google Scholar]
  20. Motoyoshi K., Suda T., Kusumoto K., Takaku F., Miura Y. Granulocyte-macrophage colony-stimulating and binding activities of purified human urinary colony-stimulating factor to murine and human bone marrow cells. Blood. 1982 Dec;60(6):1378–1386. [PubMed] [Google Scholar]
  21. Nicola N. A., Metcalf D., Johnson G. R., Burgess A. W. Separation of functionally distinct human granulocyte-macrophage colony-stimulating factors. Blood. 1979 Sep;54(3):614–627. [PubMed] [Google Scholar]
  22. Nicola N. A., Metcalf D., Matsumoto M., Johnson G. R. Purification of a factor inducing differentiation in murine myelomonocytic leukemia cells. Identification as granulocyte colony-stimulating factor. J Biol Chem. 1983 Jul 25;258(14):9017–9023. [PubMed] [Google Scholar]
  23. Okabe T., Nomura H., Sato N., Ohsawa N. Large-scale preparation and characterization of human colony-stimulating factor. J Cell Physiol. 1982 Jan;110(1):43–49. doi: 10.1002/jcp.1041100108. [DOI] [PubMed] [Google Scholar]
  24. Ruppert S., Löhr G. W., Fauser A. A. Characterization of stimulatory activity for human pluripotent stem cells (CFUGEMM). Exp Hematol. 1983 Feb;11(2):154–161. [PubMed] [Google Scholar]
  25. Salahuddin S. Z., Markham P. D., Lindner S. G., Gootenberg J., Popovic M., Hemmi H., Sarin P. S., Gallo R. C. Lymphokine production by cultured human T cells transformed by human T-cell leukemia-lymphoma virus-I. Science. 1984 Feb 17;223(4637):703–707. doi: 10.1126/science.6320367. [DOI] [PubMed] [Google Scholar]
  26. Welte K., Wang C. Y., Mertelsmann R., Venuta S., Feldman S. P., Moore M. A. Purification of human interleukin 2 to apparent homogeneity and its molecular heterogeneity. J Exp Med. 1982 Aug 1;156(2):454–464. doi: 10.1084/jem.156.2.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Westbrook C. A., Gasson J. C., Gerber S. E., Selsted M. E., Golde D. W. Purification and characterization of human T-lymphocyte-derived erythroid-potentiating activity. J Biol Chem. 1984 Aug 25;259(16):9992–9996. [PubMed] [Google Scholar]
  28. Wu M. C., Yunis A. A. Common pattern of two distinct types of colony-stimulating factor in human tissues and cultured cells. J Clin Invest. 1980 Mar;65(3):772–775. doi: 10.1172/JCI109726. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES