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Improved gene transfer into canine hematopoietic repopulating cells using CD34-enriched marrow cells in combination with a gibbon ape leukemia virus–pseudotype retroviral vector

Gene Therapy volume 6, pages 966–972 (1999)Cite this article

Abstract

We have used dogs to study gene transfer into hematopoietic stem cells, because of the applicability of results in dogs to human transplantation and the availability of canine disease models that mimic human diseases. Previously we reported successful gene transfer into canine marrow repopulating cells, however, gene transfer efficiency was low, usually below 0.1% (Kiem et al, Hum Gene Ther 1996; 7: 89). In this study we have used CD34-enriched marrow cells to study different retroviral pseudotypes for their ability to transduce canine hematopoietic repopulating cells. Cells were divided into two equal fractions that were cocultivated for 72 h with irradiated packaging cells producing vector with different retroviral pseudotypes (GALV, amphotropic or 10A1). The vectors used contained small sequence differences to allow differentiation of cells genetically marked by the different vectors. Nonadherent and adherent cells from the cultures were infused into four dogs after a myeloablative dose of 920 cGy total body irradiation. Polymerase chain reaction (PCR) analysis of DNA from peripheral blood and marrow after transplant showed that the highest gene transfer rates (up to 10%) were obtained with the GALV-pseudotype vector. Gene transfer levels have remained stable now for more than 18 months. Southern blot analysis confirmed the high gene transfer rate. Interference studies on canine D17 cells revealed that 10A1 virus behaved like an amphotropic virus and was not able to use the GALV receptor. In summary, our results show improved gene transfer into canine hematopoietic repopulating cells when CD34-enriched cells are transduced by cocultivation on a GALV-pseudotype packaging cell line in combination with a GALV-pseudotype vector. Furthermore, these results demonstrate that the monoclonal antibody to canine CD34 used in this study is able to enrich for hematopoietic repopulating cells.

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References

  1. Taylor N et al. Correction of interleukin-2 receptor function in X-SCID lymphoblastoid cells by retrovirally mediated transfer of the gamma-c gene Blood 1996 87: 3103–3107

    CAS  PubMed  Google Scholar 

  2. Shull RM et al. Bone marrow transplantation in canine mucopolysaccharidosis I: effects within the central nervous system J Clin Invest 1987 79: 435–443

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Dunbar CE et al. Improved retroviral gene transfer into murine and rhesus peripheral blood or bone marrow repopulating cells primed in vivo with stem cell factor and granulocyte colony-stimulating factor Proc Natl Acad Sci USA 1996 93: 11871–11876

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Kiem H-P et al. Gene transfer into marrow repopulating cells: comparison between amphotropic and gibbon ape leukemia virus pseudotyped retroviral vectors in a competitive repopulation assay in baboons Blood 1997 90: 4638–4645

    CAS  PubMed  Google Scholar 

  5. Kiem H-P et al. Long-term persistence of canine hematopoietic cells genetically marked by retrovirus vectors Hum Gene Ther 1996 7: 89–96

    Article  CAS  PubMed  Google Scholar 

  6. Kiem H-P et al. Retrovirus-mediated gene transduction into canine peripheral blood repopulating cells Blood 1994 83: 1467–1473.

    CAS  PubMed  Google Scholar 

  7. von Kalle C et al. Increased gene transfer into human hematopoietic progenitor cells by extended in vitro exposure to a pseudotyped retroviral vector Blood 1994 84: 2890–2897

    CAS  PubMed  Google Scholar 

  8. Bauer TR Jr, Miller AD, Hickstein DD . Improved transfer of the leukocyte integrin CD18 subunit into hematopoietic cell lines using retroviral vectors having a gibbon ape leukemia virus envelope Blood 1995 86: 2379–2387

    PubMed  Google Scholar 

  9. Bunnell BA et al. High efficiency retrovirus-mediated gene transfer into human and nonhuman primate peripheral blood lymphocytes Proc Natl Acad Sci USA 1995 92: 7739–7743

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Orlic D et al. The level of mRNA encoding the amphotropic retrovirus receptor in mouse and human hematopoietic stem cells is low and correlates with the efficiency of retrovirus transduction Proc Natl Acad Sci USA 1996 93: 11097–11102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Miller DG, Miller AD . A family of retroviruses that utilize related phosphate transporters for cell entry J Virol 1994 68: 8270–8276

    CAS  PubMed  PubMed Central  Google Scholar 

  12. McSweeney PA et al. Canine CD34: cloning of the cDNA and evaluation of an antiserum to recombinant protein Blood 1996 88: 1992–2003

    CAS  PubMed  Google Scholar 

  13. McSweeney PA et al. Characterization of monoclonal antibodies that recognize canine CD34 Blood 1998 91: 1977–1986

    CAS  PubMed  Google Scholar 

  14. Civin CI et al. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells JImmunol 1984 133: 157–165.

    CAS  Google Scholar 

  15. Andrews RG, Singer JW, Bernstein ID . Monoclonal antibody 12.8 recognizes a 115-kd molecule present on both unipotent and multipotent hematopoietic colony-forming cells and their precursors Blood 1986 67: 842–845

    CAS  PubMed  Google Scholar 

  16. Miller AD, Chen F . Retrovirus packaging cells based on 10A1 murine leukemia virus for production of vectors that use multiple receptors for cell entry J Virol 1996 70: 5564–5571

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Dick JE et al. Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/WV mice Cell 1985 42: 71–79

    Article  CAS  PubMed  Google Scholar 

  18. Nolta JA, Smogorzewska EM, Kohn DB . Analysis of optimal conditions for retrovirus-mediated transduction of primitive human hematopoietic cells Blood 1995 86: 101–110

    CAS  PubMed  Google Scholar 

  19. Moritz T et al. Fibronectin improves transduction of reconstituting hematopoietic stem cells by retroviral vectors: evidence of direct viral binding to chymotryptic carboxy-terminal fragments Blood 1996 88: 855–862

    CAS  PubMed  Google Scholar 

  20. Schuening FG et al. Retrovirus-mediated gene transduction into long-term repopulating marrow cells of dogs Blood 1991 78: 2568–2576

    CAS  PubMed  Google Scholar 

  21. Whitney KM, Goodman SA, Bailey EM, Lothrop CD Jr . The molecular basis of canine pyruvate kinase deficiency Exp Hematol 1994 22: 866–874

    CAS  PubMed  Google Scholar 

  22. Van Beusechem VW, Valerio D . Gene transfer into hematopoietic stem cells of nonhuman primates (review) Hum Gene Ther 1996 7: 1649–1668

    Article  CAS  PubMed  Google Scholar 

  23. Larochelle A et al. Identification of primitive human hematopoietic cells capable of repopulating NOD/SCID mouse bone marrow: implications for gene therapy Nature Med 1996 2: 1329–1337.

    Article  CAS  PubMed  Google Scholar 

  24. Miller AD, Buttimore C . Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production Mol Cell Biol 1986 6: 2895–2902

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Miller AD et al. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus J Virol 1991 65: 2220–2224

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Sandmaier BM et al. Biochemical characterization of a unique canine myeloid antigen Leukemia 1991 5: 125–130

    CAS  PubMed  Google Scholar 

  27. Miller DG, Edwards RH, Miller AD . Cloning of the cellular receptor for amphotropic murine retroviruses reveals homology to that for gibbon ape leukemia virus Proc Natl Acad Sci USA 1994 91: 78–82

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported in part by grants HL36444, HL03701, DK42716, and DK47754 awarded by the National Institutes of Health, DHHS, Bethesda, MD. HPK is a Markey Molecular Medicine Investigator. We thank Eric Bell, Barbara Johnston, DVM, and the staff of the Fred Hutchinson Cancer Research Center Clinical Hematology laboratory for their excellent assistance. We also wish to acknowledge the assistance of Harriet Childs in preparing the manuscript.

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Authors and Affiliations

  1. Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    H-P Kiem, P A McSweeney, B Bruno, M Goerner, G Buron, J Morris & R Storb

  2. Department of Medicine, University of Washington School of Medicine, Seattle, WA, USA

    H-P Kiem, P A McSweeney & R Storb

  3. Molecular Medicine Divisions, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    A D Miller

  4. Department of Pathology, University of Washington School of Medicine, Seattle, WA, USA

    A D Miller

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  1. H-P Kiem
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  2. P A McSweeney
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  3. B Bruno
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Kiem, HP., McSweeney, P., Bruno, B. et al. Improved gene transfer into canine hematopoietic repopulating cells using CD34-enriched marrow cells in combination with a gibbon ape leukemia virus–pseudotype retroviral vector. Gene Ther 6, 966–972 (1999). https://doi.org/10.1038/sj.gt.3300925

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