Expression of HIV-1 integrase in CEM cells inhibits HIV-1 replication.
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.
van Griensven
Johan
J
Rega Institute for Medical Research, Minderbroedersstraat 10, B-3000 Leuven, Belgium.
Zhan
Xiaojing
X
Van Maele
Bénédicte
B
Pluymers
Wim
W
Michiels
Martine
M
De Clercq
Erik
E
Cherepanov
Peter
P
Debyser
Zeger
Z
D000163:Acquired Immunodeficiency Syndrome / Q000628:therapy; D002460:Cell Line; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005813:Genes, Synthetic / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors; D019427:HIV Integrase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics*; D006801:Humans; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D014161:Transduction, Genetic; D016662:Virus Integration / Q000235:genetics; D014779:Virus Replication / Q000502:physiology*
D019427:HIV Integrase