Inhibition of HIV-1 by an anti-integrase single-chain variable fragment (SFv): delivery by SV40 provides durable protection against HIV-1 and does not require selection.
Human immunodeficiency virus type 1 (HIV-1) encodes several proteins that are packaged into virus particles. Integrase (IN) is an essential retroviral enzyme, which has been a target for developing agents to inhibit virus replication. In previous studies, we showed that intracellular expression of single-chain variable antibody fragments (SFvs) that bind IN, delivered via retroviral expression vectors, provided resistance to productive HIV-1 infection in T-lymphocytic cells. In the current studies, we evaluated simian-virus 40 (SV40) as a delivery vehicle for anti-IN therapy of HIV-1 infection. Prior work suggested that delivery using SV40 might provide a high enough level of transduction that selection of transduced cells might be unnecessary. In these studies, an SV40 expression vector was developed to deliver SFv-IN (SV(Aw)). Expression of the SFv-IN was confirmed by Western blotting and immunofluorescence staining, which showed that > 90% of SupT1 T-lymphocytic cells treated with SV(Aw) expressed the SFv-IN protein without selection. When challenged, HIV-1 replication, as measured by HIV-1 p24 antigen expression and syncytium formation, was potently inhibited in cells expressing SV40-delivered SFv-IN. Levels of inhibition of HIV-1 infection achieved using this approach were comparable to those achieved using murine leukemia virus (MLV) as a transduction vector, the major difference being that transduction using SV40 did not require selection in culture whereas transduction with MLV did require selection. Therefore, the SV40 vector as gene delivery system represents a novel therapeutic strategy for gene therapy to target HIV-1 proteins and interfere with HIV-1 replication.
BouHamdan
M
M
Dorrance H Hamilton Laboratories, Department of Medicine, Thomas Jefferson University, Philadelphia, PA, USA.
Duan
L X
LX
Pomerantz
R J
RJ
Strayer
D S
DS
D015153:Blotting, Western; D002478:Cells, Cultured; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D015658:HIV Infections / Q000628:therapy*; D019427:HIV Integrase / Q000032:analysis / Q000276:immunology; D015497:HIV-1; D006801:Humans; D007128:Immunoglobulin Fragments / Q000008:administration & dosage / Q000276:immunology; D009052:Leukemia Virus, Murine / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection / Q000379:methods; D014779:Virus Replication
D007128:Immunoglobulin Fragments; D019427:HIV Integrase