Mr. Mautino et al., Inhibition of human immunodeficiency virus type 1 (HIV-1) replication by HIV-1-based lentivirus vectors expressing transdominant Rev, J VIROLOGY, 75(8), 2001, pp. 3590-3599
Retrovirus vectors expressing transdominant-negative mutants of Rev (TdRev)
inhibit human immunodeficiency virus type 1 (HIV-1) replication by prevent
ing the nuclear export of unspliced viral transcripts, thus inhibiting the
synthesis of Gag-Pol, Env, and genomic RNA. The use of HIV-1-based vectors
to express TdRev would have the advantage of allowing access to nondividing
hematopoietic cells. It would also provide additional levels of protection
by sequestering the viral regulatory proteins Tat and Rev, competing for e
ncapsidation into wild-type virions, and inhibiting reverse transcription.
Here we describe HIV-1-based vectors that express TdRev, These vectors cont
ain mutations in the splicing signals or replacement of the Rev-responsive
element by the simian retrovirus type 1 constitutive transport element, mak
ing them less sensitive to the inhibitory effects of TdRev, In addition, ov
erexpression of Rev and the use of an HIV-1 helper plasmid that drives high
levels of Gag-Pol synthesis were used to transiently overcome the inhibiti
on by TdRev of the synthesis of Gag-Pol during vector production. SupT1 cel
ls transduced with these vectors were more resistant to HIV-1 replication t
han cells transduced with Moloney murine leukemia virus-based vectors expre
ssing TdRev, Furthermore, we show that these vectors can be mobilized by th
e wild-type virus, reducing the infectivity of virions escaping inhibition
and conferring protection against HIV-1 replication to previously untransdu
ced cells.