A new reporter system for detection of retroviral infection

Gene Ther. 1995 Aug;2(6):369-76.

Abstract

We describe a novel reporter molecule, the murine surface antigen Thy-1, useful for immunoselection and detection of retrovirus-mediated transduction by flow cytometry. A cDNA encoding the murine thy-1 gene was isolated, and cell surface expression of its gene product was demonstrated. The Thy-1 glycoprotein was tested as a cell surface reporter molecule in the context of replication-defective and -competent retroviruses. Cells transduced via murine retroviral vectors carrying the thy-1 and the neomycin phosphotransferase genes express Thy-1 glycoprotein on their surfaces. The Thy-1 marker is potentially useful in gene transfer protocols because selection of transduced cells can be achieved by immunoselection with anti-Thy-1 antibodies shortly after infection with the retroviral vector. In addition, a human immunodeficiency virus type 1 (HIV-1) recombinant expressing Thy-1 is described, which is replication-competent and syncytium-inducing in human peripheral blood mononuclear cells (PBMCs) and immortalized CD4-positive cell lines. Cells infected with this HIV-1 recombinant express Thy-1 on their surfaces and can be detected and purified by fluorescence-activated cell sorting (FACS). Because of these properties, retroviruses expressing this genetic marker can be useful for studies in gene therapy and of the retroviral life-cycle.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • CD4-Positive T-Lymphocytes / virology
  • Cell Line
  • Cells, Cultured
  • Chlorocebus aethiops
  • DNA Primers
  • DNA, Viral / analysis
  • DNA, Viral / biosynthesis*
  • Flow Cytometry
  • Gene Expression
  • Giant Cells
  • HIV Core Protein p24 / biosynthesis
  • HIV Long Terminal Repeat
  • HIV-1 / genetics*
  • Humans
  • Kanamycin Kinase
  • Lymphocytes / virology*
  • Mice
  • Molecular Sequence Data
  • Phosphotransferases (Alcohol Group Acceptor) / biosynthesis
  • Polymerase Chain Reaction
  • Thy-1 Antigens / analysis
  • Thy-1 Antigens / biosynthesis*
  • Transfection*
  • Tumor Cells, Cultured

Substances

  • DNA Primers
  • DNA, Viral
  • HIV Core Protein p24
  • Thy-1 Antigens
  • Phosphotransferases (Alcohol Group Acceptor)
  • Kanamycin Kinase