Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors

D. S. Strayer, R. J. Pomerantz, M. Yu, M. Rosenzweig, M. BouHamdan, S. Yurasov, R. P. Johnson, Harris Goldstein

Research output: Contribution to journalArticle

31 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)886-895
Number of pages10
JournalGene Therapy
Volume7
Issue number10
StatePublished - May 2000

Fingerprint

Hematopoietic Stem Cells
Genes
Bone Marrow
Transgenes
Polymerase Chain Reaction
Hepatitis B Surface Antigens
Bone Marrow Cells
Immunochemistry
Single-Chain Antibodies
DNA Primers
SCID Mice
Replantation
Macaca mulatta
Primates
Stem Cells
Cell Culture Techniques
Genome
T-Lymphocytes
DNA

Keywords

  • Bone marrow
  • CD34 cells
  • Gene theapy
  • SV40

ASJC Scopus subject areas

  • Genetics

Cite this

Strayer, D. S., Pomerantz, R. J., Yu, M., Rosenzweig, M., BouHamdan, M., Yurasov, S., ... Goldstein, H. (2000). Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors. Gene Therapy, 7(10), 886-895.

Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors. / Strayer, D. S.; Pomerantz, R. J.; Yu, M.; Rosenzweig, M.; BouHamdan, M.; Yurasov, S.; Johnson, R. P.; Goldstein, Harris.

In: Gene Therapy, Vol. 7, No. 10, 05.2000, p. 886-895.

Research output: Contribution to journalArticle

Strayer, DS, Pomerantz, RJ, Yu, M, Rosenzweig, M, BouHamdan, M, Yurasov, S, Johnson, RP & Goldstein, H 2000, 'Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors', Gene Therapy, vol. 7, no. 10, pp. 886-895.
Strayer DS, Pomerantz RJ, Yu M, Rosenzweig M, BouHamdan M, Yurasov S et al. Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors. Gene Therapy. 2000 May;7(10):886-895.
Strayer, D. S. ; Pomerantz, R. J. ; Yu, M. ; Rosenzweig, M. ; BouHamdan, M. ; Yurasov, S. ; Johnson, R. P. ; Goldstein, Harris. / Efficient gene transfer to hematopoietic progenitor cells using SV40-derived vectors. In: Gene Therapy. 2000 ; Vol. 7, No. 10. pp. 886-895.
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