Abstract
The central role of endoconvertases and HIV-1 protease (HIV-1 PR) in the processing of HIV proproteins makes the design of specific inhibitors important in anti-HIV gene therapy. Accordingly, we tested native α1 antitrypsin (α1AT) delivered by a recombinant simian virus-40-based vector, SV(AT), as an inhibitor of HIV-1 proprotein maturation. Cell lines and primary human lymphocytes were transduced with SV(AT) without selection and detectable toxicity. Expression of α1AT was confirmed by Northern blotting, immunoprecipitation and immunostaining. SV(AT)-transduced cells showed no evidence of HIV-1-related cytopathic effects when challenged with high doses of HIV-1NL4-3. As measured by HIV-1 p24 assay, SV(AT)-transduced cells were protected from HIV-1NL4-3 at challenge dose of 40 000 TCID50 (MOI = 0.04). In addition, peripheral blood lymphocytes treated with SV(AT) were protected from HIV doses challenge up to 40 000 TCID50 (MOI = 0.04). By Western blot analyses, the delivered α1AT inhibited cellular processing of gp 160 to gp 120 and decreased HIV-1 virion gp 120. SV(AT) inhibited processing of p55Gag as well. Furthermore, high levels of uncleaved p55Gag protein were detected in HIV virus particles recovered from SV(AT)-transduced cells lines and primary lymphocytes. Thus, delivering α1AT using SV(AT) to human lymphocytes strongly inhibits replication of HIV-1, most likely by inhibiting the activities both of the cellular serine proteases involved in processing gp160 and of the aspartyl protease, HIV-1 PR, which cleaves p55gag, α1AT delivered by SV(AT) may represent a novel and effective strategy for gene therapy to interfere with HIV replication, by blocking a stage in the virus replicative cycle that has until now been inaccessible to gene therapeutic intervention.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 467-477 |
| Number of pages | 11 |
| Journal | Gene Therapy |
| Volume | 10 |
| Issue number | 6 |
| DOIs | |
| State | Published - Mar 2003 |
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Keywords
- α
- Antitrypsin
- gp160
- p55
- SV40
ASJC Scopus subject areas
- Genetics
Cite this
HIV-1 proprotein processing as a target for gene therapy. / Cordelier, P.; Zern, Mark A; Strayer, D. S.
In: Gene Therapy, Vol. 10, No. 6, 03.2003, p. 467-477.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - HIV-1 proprotein processing as a target for gene therapy
AU - Cordelier, P.
AU - Zern, Mark A
AU - Strayer, D. S.
PY - 2003/3
Y1 - 2003/3
N2 - The central role of endoconvertases and HIV-1 protease (HIV-1 PR) in the processing of HIV proproteins makes the design of specific inhibitors important in anti-HIV gene therapy. Accordingly, we tested native α1 antitrypsin (α1AT) delivered by a recombinant simian virus-40-based vector, SV(AT), as an inhibitor of HIV-1 proprotein maturation. Cell lines and primary human lymphocytes were transduced with SV(AT) without selection and detectable toxicity. Expression of α1AT was confirmed by Northern blotting, immunoprecipitation and immunostaining. SV(AT)-transduced cells showed no evidence of HIV-1-related cytopathic effects when challenged with high doses of HIV-1NL4-3. As measured by HIV-1 p24 assay, SV(AT)-transduced cells were protected from HIV-1NL4-3 at challenge dose of 40 000 TCID50 (MOI = 0.04). In addition, peripheral blood lymphocytes treated with SV(AT) were protected from HIV doses challenge up to 40 000 TCID50 (MOI = 0.04). By Western blot analyses, the delivered α1AT inhibited cellular processing of gp 160 to gp 120 and decreased HIV-1 virion gp 120. SV(AT) inhibited processing of p55Gag as well. Furthermore, high levels of uncleaved p55Gag protein were detected in HIV virus particles recovered from SV(AT)-transduced cells lines and primary lymphocytes. Thus, delivering α1AT using SV(AT) to human lymphocytes strongly inhibits replication of HIV-1, most likely by inhibiting the activities both of the cellular serine proteases involved in processing gp160 and of the aspartyl protease, HIV-1 PR, which cleaves p55gag, α1AT delivered by SV(AT) may represent a novel and effective strategy for gene therapy to interfere with HIV replication, by blocking a stage in the virus replicative cycle that has until now been inaccessible to gene therapeutic intervention.
AB - The central role of endoconvertases and HIV-1 protease (HIV-1 PR) in the processing of HIV proproteins makes the design of specific inhibitors important in anti-HIV gene therapy. Accordingly, we tested native α1 antitrypsin (α1AT) delivered by a recombinant simian virus-40-based vector, SV(AT), as an inhibitor of HIV-1 proprotein maturation. Cell lines and primary human lymphocytes were transduced with SV(AT) without selection and detectable toxicity. Expression of α1AT was confirmed by Northern blotting, immunoprecipitation and immunostaining. SV(AT)-transduced cells showed no evidence of HIV-1-related cytopathic effects when challenged with high doses of HIV-1NL4-3. As measured by HIV-1 p24 assay, SV(AT)-transduced cells were protected from HIV-1NL4-3 at challenge dose of 40 000 TCID50 (MOI = 0.04). In addition, peripheral blood lymphocytes treated with SV(AT) were protected from HIV doses challenge up to 40 000 TCID50 (MOI = 0.04). By Western blot analyses, the delivered α1AT inhibited cellular processing of gp 160 to gp 120 and decreased HIV-1 virion gp 120. SV(AT) inhibited processing of p55Gag as well. Furthermore, high levels of uncleaved p55Gag protein were detected in HIV virus particles recovered from SV(AT)-transduced cells lines and primary lymphocytes. Thus, delivering α1AT using SV(AT) to human lymphocytes strongly inhibits replication of HIV-1, most likely by inhibiting the activities both of the cellular serine proteases involved in processing gp160 and of the aspartyl protease, HIV-1 PR, which cleaves p55gag, α1AT delivered by SV(AT) may represent a novel and effective strategy for gene therapy to interfere with HIV replication, by blocking a stage in the virus replicative cycle that has until now been inaccessible to gene therapeutic intervention.
KW - α
KW - Antitrypsin
KW - gp160
KW - p55
KW - SV40
UR - http://www.scopus.com/inward/record.url?scp=0344642957&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0344642957&partnerID=8YFLogxK
U2 - 10.1038/sj.gt.3301891
DO - 10.1038/sj.gt.3301891
M3 - Article
C2 - 12621451
AN - SCOPUS:0344642957
VL - 10
SP - 467
EP - 477
JO - Gene Therapy
JF - Gene Therapy
SN - 0969-7128
IS - 6
ER -