Advertisement

Effects of Leukemia Inhibitory Factor (LIF) on Gene Transfer Efficiency into Murine Hematolymphoid Progenitors

  • Frederick A. Fletcher
  • Kateri A. Moore
  • Douglas E. Williams
  • Dirk Anderson
  • Charles Maliszewski
  • John W. Belmont
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 292)

Abstract

We have investigated the effects of the cytokine leukemia inhibitory factor (LIF) on recovery and retroviral infection of murine hematopoietic stem cells maintained in short-term culture. Up to a two-fold increase in CFU-S13 recovery was observed, from 9.7 x 10-5 cells in untreated controls to 17.6 x 10-5 cells when lOU/ml LIF is added to the culture medium. Intermediate concentrations of LIF (.lU/ml and lU/ml) were not significantly different from the control. Histological analysis of spleen colonies harvested thirteen days posttransplant demonstrated that LIF does not cause a detectable alteration in the differentiative potential of CFU-S13. The efficiency of retroviral-vector infection in CFU-S13 is also improved, from 15% (24/158) in untreated controls to 91% (116/127) at a LIF concentration of l0U/ml. LIF concentrations of .lU/ml and lU/ml increased infection efficiency to 35% (14/40) and 71% (37/51), respectively. Analysis of proviral insertion sites in spleen colonies indicated that some CFU-S13 precursors were infected in the LIF-treated marrows, but no identical pairs were detected in the controls. Finally, long-term expression of provirally-encoded human adenosine deaminase (hADA) was measured in hematopoietic tissues of bone marrow transplant recipients six months posttransplant. In all tissues analyzed (spleen, thymus, bone marrow, splenic B cells, peritoneal macrophages, and blood) differentiated progeny of LIF-treated marrows had higher levels of hADA than untreated controls. Tenfold increases in levels of hADA are detected in some tissues, but levels were variable. These experiments demonstrate that LIF directly or indirectly enhances retroviral infection efficiency of hematopoietic stem cells, and might be used to improved existing gene transfer protocols.

Keywords

Hematopoietic Stem Cell Leukemia Inhibitory Factor Infection Efficiency Hematopoietic Tissue Myeloid Leukemic Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    J. W. Belmont, G. R. MacGregor, K. Wager-Smith, F. A. Fletcher, K. A. Moore, D. Hawkins, D. Villalon, S. M.-W. Chang, and C. T. Caskey, Expression of human adenosine deaminase in murine hematopoietic cells, Mol. Cell Biol. 8:5116 (1988).PubMedGoogle Scholar
  2. 2.
    K. A. Moore, F. A. Fletcher, D. K. Villalon, A. E. Utter, and J. W. Belmont, Human adenosine deaminase expression in mice, Blood 75:2085 (1990).PubMedGoogle Scholar
  3. 3.
    L. G. Lajtha, Stem cell concepts, Differentiation 14:23 (1979).PubMedCrossRefGoogle Scholar
  4. 4.
    D. M. Bodine, S. Karlsson, and A. W. Nienhuis, Combination of interleukins 3 and 6 preserves stem cell function in culture and enhances retrovirus-mediated gene transfer into hematopoietic stem cells, Proc. Natl. Acad. Sci. USA 86:8897 (1989).PubMedCrossRefGoogle Scholar
  5. 5.
    D. J. Hilton, N. A. Nicola, N. M. Gough, and D. Metcalf, Resolution and purification of three distinct factors produced by Krebs ascites cells which have differentiation-including activity on murine myeloid leukemic cell lines, J. Biol. Chem 263:9238 (1988).PubMedGoogle Scholar
  6. 6.
    D. Metcalf, Actions and interactions of G-CSF, LIF, and IL-6 on normal and leukemic murine cells, Leukemia 3:349 (1989).PubMedGoogle Scholar
  7. 7.
    A. Godard, H. Gascan, J. Naulet, M.-A. Peyrat, Y. Jacques, J.-P. Soulillou, and J.-F. Moreau, Biochemical characterization and purification of HILDA, a human lymphokine active on eosinophils and bone marrow cells, Blood 71:1618 (1988).PubMedGoogle Scholar
  8. 8.
    D. J. Hilton, N. A. Nicola, and D. Metcalf, Specific binding of murine leukemia inhibitory factor to normal and leukemic monocytic cells, Proc. Natl. Acad. Sci. USA 85:5971 (1988).PubMedCrossRefGoogle Scholar
  9. 9.
    A. G. Smith, J. K. Heath, D. D. Donaldson, G. G. Wong, J. Moreau, M. Stahl, and D. Rogers, Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides, Nature 336:688 (1988).PubMedCrossRefGoogle Scholar
  10. 10.
    R. L. Williams, D. J. Hilton, S. Pease, T. A. Willson, C. L. Stewart, D. P. Gearing, E. F. Wagner, D. Metcalf, N. A. Nicola, and N. M. Gough, Myeloid leukemia inhibitory factor maintains the developmental potential of embryonic stem cells, Nature 336:684 (1988).PubMedCrossRefGoogle Scholar
  11. 11.
    S. C. Kogan, M. Doherty, and J. Gitschier, An improved method for prenatal diagnosis of genetic diseases by amplified DNA sequences, N. Engl. J. Med 317:985 (1987).PubMedCrossRefGoogle Scholar
  12. 12.
    E. M. Southern, Detection of specific sequences among DNA fragments separated by gel electrophoresis, J. Mol. Biol. 98:503 (1975).PubMedCrossRefGoogle Scholar
  13. 13.
    G. S. Adrian, D. A. Wiginton, and J. J. Hutton, Structure of adenosine deaminase mrnas from normal and adenosine deaminase deficient human cell lines, Mol. Cell. Biol. 4:1712 (1984).PubMedGoogle Scholar
  14. 14.
    D. Metcalf, T. Maekawa, D. Hilton, N. Nicola, D. Gearing, and N. Gough, Interactions of leukemia inhibitory factor (LIF) IL-6 and colony-stimulating factors on murine and human leukemic cells. Exp. Hematol. 17:483 (1989).Google Scholar
  15. 15.
    J. Harel, E. Rassart, and P. Jolicoeur, Cell cycle dependence of synthesis of unintegrated viral DNA in mouse cells newly infected with muine leukemia virus, Virology 110:202 (1981).PubMedCrossRefGoogle Scholar
  16. 16.
    I. S. Y. Chen, and H. Temin, Establishment of infection by spleen necrosis virus: inhibition in stationary cells and the role of secondary infection, J, Virol. 41:183 (1982).Google Scholar
  17. 17.
    D. E. Williams, A. E. Namen, D. Y. Mochizuki, and R. W. Overell, Clonal growth of murine pre-B colony forming cells and their targeted infection by a retroviral vector: dependence on interleukin-7, Blood 75:1132 (1990),PubMedGoogle Scholar
  18. 18.
    G. M. Springett, R. C. Moen, S. Anderson, R. M. Blaese, and W. F. Anderson, Infection efficiency of T lymphocytes with amphotropic retroviral vectors is cell cycle dependent, J. Virol 63:3865 (1989).PubMedGoogle Scholar
  19. 19.
    A. J. Becker, E. A. Mc-Culloch, L. Siminovitch, and J. E. Till, The effect of different demands for blood cell production on DNA synthesis by haemopoietic colony forming cells of mice, Blood 26:296 (1965).PubMedGoogle Scholar
  20. 20.
    B. I. Lord, L. G. Lajtha, and J. Gidali, Measurement of the kinetic status of bone marrow precursor cells: three cautionary tales, Cell and Tissue Kin 7:507 (1974).Google Scholar
  21. 21.
    A. G. Leary, G. C. Wong, S. C. Clark, and M. Ogawa, Leukemia inhibitory factor (LIF)/differentiation-inducing activity (DIA) is a synergistic factor for human hemopoietic stem cells, Exp. Hematol. 17:524 (1989).Google Scholar
  22. 22.
    J. Ikebuchi, G. G. Wong, S. C. Clark, J. N. Ihle, Y. Hirai, and M. Ogawa, Interleukin 6 enhancement of interleukin 3-dependent proliferation of multi-potential hemopoietic progenitors, Proc. Natl. Acad. Sci. USA 84:9035 (1987).PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Frederick A. Fletcher
    • 2
  • Kateri A. Moore
    • 2
  • Douglas E. Williams
    • 3
  • Dirk Anderson
    • 3
  • Charles Maliszewski
    • 3
  • John W. Belmont
    • 1
    • 2
  1. 1.Howard Hughes Medical InstituteUSA
  2. 2.Institute for Molecular GeneticsBaylor College of MedicineHoustonUSA
  3. 3.Immunex Research and Development CorporationSeattleUSA

Personalised recommendations