Efficacy and safety analyses of a recombinant human immunodeficiency virus type 1 derived vector system

L. J. Chang, V. Urlacher, T. Iwakuma, Yan Cui, J. Zucali

Research output: Contribution to journalArticle

170 Citations (Scopus)

Abstract

Lentiviruses infect both dividing and nondividing cells. In this study we characterized a lentiviral vector system consisting of a packaging vector (pHP) and a transducing vector (pTV) derived from a recombinant human immunodeficiency virus type 1 (HIV-1). In pHP, the long terminal repeats (LTRs), the 5' untranslated leader and portions of the env and nef genes were deleted. The leader sequence of pHP was substituted with a modified Rous sarcoma virus (RSV) 59 bp leader containing a mutated RSV gag AUG and a functional 5' splice site. The pHP construct was found to direct Gag-Pol synthesis as efficiently as wild-type HIV-1. The pTV construct contains sequences required for RNA packaging, reverse transcription and integration, but lacks viral genes. Co-transfection of pHP, pTV and a vesicular stomatitis virus G (VSV-G) envelope plasmid produced vectors at titers of 10 5-10 6 transducing units per milliliter in 48 h. Replication-competent virus (RCV) was not detected when deletions were made in the env gene in pHP. The ability of this vector system to transduce dividing and nondividing cell in vitro and in vivo was also demonstrated. Compared with Moloney murine leukemia virus (MLV) vector, the HP/TV vectors transduced human muscle-, kidney-, liver-derived cell lines and CD34 + primary hematopoietic progenitor cells more efficiently. Although the levels of the pTV transgene expression were high soon after transduction, the expression tended to decrease with time due either to the loss of proviral DNA or to the inactivation of promoter activity, which was found to be cell type-dependent. Analyses of extrachromosomal DNA showed that the unintegrated proviral DNA of lentiviral vectors survived much longer than that of the retroviral vectors. We demonstrate that the HP/TV vector is capable of high efficiency transduction and that long-term expression of lentiviral vectors is dependent on target cell type, the internal promoter and the transgene itself in the transducing vector.

Original languageEnglish (US)
Pages (from-to)715-728
Number of pages14
JournalGene Therapy
Volume6
Issue number5
DOIs
StatePublished - May 1 1999

Fingerprint

HIV-1
env Genes
Safety
Rous sarcoma virus
Product Packaging
Transgenes
DNA
nef Genes
Moloney murine leukemia virus
Lentivirus
Vesicular Stomatitis
RNA Splice Sites
Viral Genes
Terminal Repeat Sequences
Virus Replication
Hematopoietic Stem Cells
Reverse Transcription
Transfection
Plasmids
Viruses

Keywords

  • HIV
  • Lentivirus
  • MLV
  • VSV-G
  • Vector

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Efficacy and safety analyses of a recombinant human immunodeficiency virus type 1 derived vector system. / Chang, L. J.; Urlacher, V.; Iwakuma, T.; Cui, Yan; Zucali, J.

In: Gene Therapy, Vol. 6, No. 5, 01.05.1999, p. 715-728.

Research output: Contribution to journalArticle

Chang, L. J. ; Urlacher, V. ; Iwakuma, T. ; Cui, Yan ; Zucali, J. / Efficacy and safety analyses of a recombinant human immunodeficiency virus type 1 derived vector system. In: Gene Therapy. 1999 ; Vol. 6, No. 5. pp. 715-728.
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