Transformation studies with a human T-cell leukemia virus type 1 molecular clone.
In in vitro studies human T-cell leukemia virus type 1 (HTLV-1) may be produced by stable or transient transfection of target cells with an infectious molecular clone. Studies using primary human T cells, the natural targets of HTLV-1 infection, are hampered by difficulty in achieving significant infection with cell-free virus and a poor efficiency of transfection of primary cells. A method is described for the generation of stable cell lines expressing HTLV-1 from an infectious proviral clone. The stably transfected cells can be irradiated and cocultured with human peripheral blood mononuclear cells (PBMC) resulting in infected primary cells. These cells become immortalized, IL-2 dependent lines, which contain integrated copies of provirus and express a full spectrum of viral proteins. Analysis of cellular markers indicates that immortalized cell lines consist of CD3+/CD4+ T cells, matching the most common adult T-cell leukemia (ATL) cell phenotype. The method described has great utility in the study of the replication and transformation capacity of HTLV and HTLV mutant viruses in their natural targets, primary human T lymphocytes.
Anderson
Matthew D
MD
Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA.
Ye
Jianxin
J
Xie
Li
L
Green
Patrick L
PL
D001483:Base Sequence; D002472:Cell Transformation, Viral; D003001:Cloning, Molecular / Q000379:methods; D018920:Coculture Techniques; D017931:DNA Primers; D015368:Human T-lymphotropic virus 1 / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D013601:T-Lymphocytes / Q000528:radiation effects / Q000821:virology*
D017931:DNA Primers