A new reporter system for detection of retroviral infection.
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.
Planelles
V
V
Department of Medicine, UCLA School of Medicine, USA.
Haislip
A
A
Withers-Ward
E S
ES
Stewart
S A
SA
Xie
Y
Y
Shah
N P
NP
Chen
I S
IS
D000818:Animals; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D002460:Cell Line; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis*; D005434:Flow Cytometry; D015870:Gene Expression; D015726:Giant Cells; D016655:HIV Core Protein p24 / Q000096:biosynthesis; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D019868:Kanamycin Kinase; D008214:Lymphocytes / Q000821:virology*; D051379:Mice; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D018800:Thy-1 Antigens / Q000032:analysis / Q000096:biosynthesis*; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D004279:DNA, Viral; D016655:HIV Core Protein p24; D018800:Thy-1 Antigens; D017853:Phosphotransferases (Alcohol Group Acceptor); D019868:Kanamycin Kinase