Efficient transduction by an amphotropic retrovirus vector is dependent on high-level expression of the cell surface virus receptor

Peter Kurre, Hans Peter Kiem, Julia Morris, Scott Heyward, Jean Luc Battini, A. Dusty Miller

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Transduction by murine leukemia virus-based retrovirus vectors is limited in certain cell types, particularly in nondividing cells. But transduction can be inefficient even in cells that divide rapidly. For example, exposure of 208F rat embryo fibroblasts to an excess of an amphotropic retrovirus vector encoding alkaline phosphatase results in a transduction efficiency of only about 10%, even though these cells divide rapidly. Here we show that transduction of 208F cells is limited by cell surface retrovirus receptor levels; overexpression of the amphotropic retrovirus receptor Pit2 markedly improved the transduction efficiency to 50%. To characterize receptor levels and binding affinity, we synthesized a fusion protein that joins the amino terminus of the amphotropic envelope protein to the Fc region of a human immunoglobulin G1 molecule for use in binding assays. In comparison to the parental cell line, the modified cell line showed an order of magnitude increase in binding sites of from 18,000 to 150,000 per cell. Thus, efficient transduction by an amphotropic retrovirus vector requires high-level expression of the retrovirus receptor Pit2. These results provide the rationale for further examination of the role of receptor levels in inefficient transduction, especially with regard to target cells for gene therapy, where a high transduction rate is often crucial.

Original languageEnglish (US)
Pages (from-to)495-500
Number of pages6
JournalJournal of Virology
Volume73
Issue number1
StatePublished - 1999
Externally publishedYes

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Retroviridae
Virus Receptors
Cell Surface Receptors
receptors
cells
Cell Line
Murine Leukemia Viruses
cell lines
Murine leukemia virus
Cell- and Tissue-Based Therapy
Genetic Therapy
gene therapy
Alkaline Phosphatase
virus receptors
Immunoglobulins
Proteins
Embryonic Structures
Fibroblasts
Binding Sites
immunoglobulins

ASJC Scopus subject areas

  • Immunology

Cite this

Efficient transduction by an amphotropic retrovirus vector is dependent on high-level expression of the cell surface virus receptor. / Kurre, Peter; Kiem, Hans Peter; Morris, Julia; Heyward, Scott; Battini, Jean Luc; Dusty Miller, A.

In: Journal of Virology, Vol. 73, No. 1, 1999, p. 495-500.

Research output: Contribution to journalArticle

Kurre, Peter ; Kiem, Hans Peter ; Morris, Julia ; Heyward, Scott ; Battini, Jean Luc ; Dusty Miller, A. / Efficient transduction by an amphotropic retrovirus vector is dependent on high-level expression of the cell surface virus receptor. In: Journal of Virology. 1999 ; Vol. 73, No. 1. pp. 495-500.
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