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Expression of HIV‐1 integrase in CEM cells inhibits HIV‐1 replication
Author(s) -
van Griensven Johan,
Zhan Xiaojing,
Van Maele Bénédicte,
Pluymers Wim,
Michiels Martine,
De Clercq Erik,
Cherepanov Peter,
Debyser Zeger
Publication year - 2004
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.520
Subject(s) - viral replication , transduction (biophysics) , biology , cell culture , integrase , microbiology and biotechnology , transfection , gene expression , hiv long terminal repeat , virology , gene , long terminal repeat , human immunodeficiency virus (hiv) , genetics , biochemistry
Background HIV‐1 integrase (IN) is an interesting target for the gene therapy of AIDS. Although the in vivo functions are not well characterized, it is thought that IN has pleiotropic effects and plays a central role in the interplay between the virus and the host cell. Expression of IN in mammalian cells has proven difficult. We have previously established a 293T‐derived cell line that stably expresses high levels of HIV‐1 IN from a synthetic gene. We now have constructed CEM‐derived cell lines stably expressing the enzyme or its different domains and studied the impact of IN expression on HIV‐1 replication. Methods The CEM cell lines were selected following transduction with a retroviral vector encoding the full‐length IN, the N‐terminal domain, the catalytic core or the C‐terminal domain. Stable IN expression in CEM cell lines was verified by Western blotting. The impact of IN expression on HIV‐1 replication and HIV‐1 vector transduction was studied. Results A marked inhibitory effect on HIV‐1 replication was observed in CEM cells expressing IN. Expression of IN interfered with both particle production and integration. Expression of the N‐terminal domain alone was sufficient for the inhibiting of HIV‐1 replication. Conclusions Expression of IN in CEM cells inhibits HIV‐1 replication by a cumulative inhibitory effect on integration and particle production, in accord with the known pleiotropic interactions of IN. The inhibition of HIV‐1 replication in CEM cells expressing the N‐terminal domain of IN may lead to a novel approach for the gene therapy of AIDS. Copyright © 2004 John Wiley & Sons, Ltd.