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  • Viral Transfer Technology
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Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors

Gene Therapy volume 7, pages 886–895 (2000)Cite this article

Abstract

We used recombinant SV40 (rSV40)-derived vectors to deliver transgenes to human and simian hematopoietic progenitor cells in culture, and in vivo after transduction ex vivo. rSV40 are highly efficient vectors that are made in very high titers. They infect almost all cells, whether resting or dividing. Two rSV40s were used: SV(HBS), carrying hepatitis B surface antigen as a marker; and SV(Aw) carrying IN#33, a single chain Fv antibody against HIV-1 integrase. CD34+ cells derived from human fetal bone marrow (HFBM) and rhesus macaque bone marrow were transduced once with SV(HBS) without selection. On average 60% of colonies derived from transduced CD34+ cells carried and expressed HBsAg, as assessed by PCR and immunochemistry. Transgene carriage persisted following differentiation of transduced rhesus CD34+ cells into T lymphocytes. In an effort to increase the percentage of gene-marked cells, three sequential treatments of CD34+cells were done using sv(aw), without selection. two weeks later, >95% of colonies expressed IN#33. Unselected SV(Aw)-transduced CD34+ cells from hfbm were transplanted into sublethally irradiated scid mice. bone marrow harvested 3 months later showed that >50% of bone marrow cells expressed IN#33. This is comparable with the percentage of human cells in these animals’ bone marrow as judged by immunostaining for human CD45. The stability and longevity of transduction in this setting suggests that rSV40 vectors integrate into the cellular genome. This possibility was supported by finding that PCR of genomic DNA using primer pairs with one cellular and one viral primer yielded PCR products only in transduced, but not control, cells. These PCR products hybridized with an SV40 DNA fragment. Thus, rSV40 vectors transduce normal human and primate bone marrow progenitor cells effectively without selection, and maintain transgene expression in vivo following reimplantation. Such high efficiency transduction may be useful in treating diseases of CD34+ cells and their derivatives.

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Acknowledgements

This work was made possible by the excellent technical support provided by Joe Milano and Danlan Wei. The services of the Immunopathology Laboratory at Jefferson University Hospital, under the direction of Dr Roland Schwarting and Mr Al Kovatich, are gratefully acknowledged. These studies were supported by USPHS grants AI41399, RR13156, RR00168 and CA73473.

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Authors and Affiliations

  1. Department of Pathology, Jefferson Medical College, Philadelphia, PA, USA

    D S Strayer & M Yu

  2. Division of Infectious Disease, Department of Medicine, Jefferson Medical College, Philadelphia, PA, USA

    R J Pomerantz & M BouHamdan

  3. Division of Infectious Diseases, Department of Medicine, Harvard Medical College and the New England Regional Primate Research Center, Boston, MA, USA

    M Rosenzweig & R P Johnson

  4. Division of Immunology, Department of Pediatrics, Albert Einstein College of Medicine, Bronx, NY, USA

    S Yurasov & H Goldstein

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  1. D S Strayer
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  2. R J Pomerantz
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  3. M Yu
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  4. M Rosenzweig
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  6. S Yurasov
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Strayer, D., Pomerantz, R., Yu, M. et al. Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors. Gene Ther 7, 886–895 (2000). https://doi.org/10.1038/sj.gt.3301159

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