Blut

, Volume 52, Issue 1, pp 51–58 | Cite as

Identification of leukemia cell-derived inhibitory activity (LIA) in conditioned media from human myeloid leukemic cell line ML-2

  • V. Cukrová
  • Z. Hrkal
  • H. Kopřivová
  • J. Neuwirt
Original Articles

Summary

Conditioned media from the human myeloid leukemic cell line ML-2 contain a factor that inhibits the entry of normal CFU-GM into S phase of mitotic cycle as measured by the 3H-TdR suicide technique. This factor was detected in conditioned media prepared by incubating 5×106 ML-2 cells/ml or 1×106 ML-2 cells/ml in serum-free RPMI for 5 or 24 hours respectively, and was isolated by ultrafiltration through an XM300 Diaflo membrane followed by chromatography on Sepharose 6B. Ferritin, prepared from human placenta, had the same inhibitory effect on CFU-GM. Antibodies against human placental ferritin completely inactivated the inhibitory effect of both human placental ferritin and the factor released from ML-2 cells. The inhibitory activity produced by the cell-line ML-2 was considered as LIA (leukemia cell-derived inhibitory activity) earlier found in HL-60 cell line and AML and CML cells.

Key words

LIA Leukemia cell-derived inhibitory activity CM Conditioned medium Granulopoiesis CFU-GM Progenitor cells of granulocytes and macrophages 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Bognacki J, Broxmeyer HE, Lobue J (1981) Isolation and biochemical characterization of leukemia associated inhibitory activity that suppresses colony and cluster formation of cells. Biochim Biophys Acta 672: 176–190Google Scholar
  2. 2.
    Broxmeyer HE, Jacobsen N, Kurland J, Mendelsohn N, Moore MAS (1978) In vitro suppression of normal granulocytic stem cells by inhibitory activity derived from human leukemia cells. J Nat Cancer Inst 60: 497–511Google Scholar
  3. 3.
    Broxmeyer HE, Bognacki J, Dorner MH, de Sousa M (1981) Identification of leukemia-associated inhibitory activity as acidic isoferritin. A regulatory role for acidic isoferritin in the production of granulocytes and macrophages. J Exp Med 153: 1426–1444Google Scholar
  4. 4.
    Broxmeyer HE (1982) Relationship of cell-cycle expression of Ia-like antigenic determinants on normal and leukemic human granulocyte-macrophage progenitor cells to regulation in vitro by acidic iso-ferritins. J Clin Invest 69: 632–642Google Scholar
  5. 5.
    Broxmeyer HE, Bognacki J, Ralph P, Dorner MH, Lu L, Castro-Malaspina H (1982) Monocyte-macrophage derived acidic isoferritins: normal feedback regulators of granulocyte-macrophage progenitor cells. Blood 60: 595–607Google Scholar
  6. 6.
    Broxmeyer HE (1983) Acid isoferritin inhibitory activity. Blood 62: 702–703Google Scholar
  7. 7.
    Broxmeyer HE, Gentile P, Bognacki J, Ralph P (1983) Lactoferrin, transferrin and acidic isoferritins: regulatory molecules with potential therapeutic value in leukemia. Blood Cells 9: 83–105Google Scholar
  8. 8.
    Drysdale JW, Munro HN (1965) Small-scale isolation of ferritin for the assay of the incorporation of 14C-labelled amino acids. Biochem J 95: 851–858Google Scholar
  9. 9.
    Eckersall D, Conner JG (1984) The prevention of distortion in ultrathin-layer polyacrylamide gel isoelectric focusing. Anal Biochem 138: 52–58Google Scholar
  10. 10.
    Minowada J, Sagawa K, Trowbrodge IS, Kung PD, Goldstein G (1982) Markerprofiles of 55 human leukemia-lymphoma cell lines. In: Kaplan HS, Rosenberg SA (eds) Malignant lymphomas. Academic Press, New York, p 53Google Scholar
  11. 11.
    Olofsson T, Olsson I (1980) Suppression of normal granulopoiesis in vitro by a leukemia associated inhibitor (LAI) derived from human prolyelotic cell line (HL-60). Leuk Res 4: 437–447Google Scholar
  12. 12.
    Olofsson T, Olsson I (1980) Biochemical characterization of a leukemia-associated inhibitor (LAI) suppressing normal granulopoiesis in vitro. Blood 55: 983–991Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • V. Cukrová
    • 1
  • Z. Hrkal
    • 1
  • H. Kopřivová
    • 2
  • J. Neuwirt
    • 1
  1. 1.Institute of Hematology and Blood TransfusionPrague 2Czechoslovakia
  2. 2.Institute of Biophysics and Nuclear MedicineCharles UniversityPrague 2Czechoslovakia

Personalised recommendations