Inhibition of clinical human immunodeficiency virus (HIV) type 1 isolates in primary CD4+ T lymphocytes by retroviral vectors expressing anti-HIV genes

Abstract

Gene therapy may be of benefit in human immunodeficiency virus type 1 (HIV-1)-infected individuals by virtue of its ability to inhibit virus replication and prevent viral gene expression. It is not known whether anti-HIV-1 gene therapy strategies based on antisense or transdominant HIV-1 mutant proteins can inhibit the replication and expression of clinical HIV-1 isolates in primary CD4 ؉ T lymphocytes. We therefore transduced CD4 ؉ T lymphocytes from uninfected individuals with retroviral vectors expressing either HIV-1-specific antisense-TAR or antisense-Tat/Rev RNA, transdominant HIV-1 Rev protein, and a combination of antisense-TAR and transdominant Rev. The engineered CD4 ؉ T lymphocytes were then infected with four different clinical HIV-1 isolates. We found that replication of all HIV-1 isolates was inhibited by all the anti-HIV vectors tested. Greater inhibition of HIV-1 was observed with transdominantRev than with antisense RNA. We hereby demonstrated effective protection by antisense RNA or transdominant mutant proteins against HIV-1 infection in primary CD4 ؉ T lymphocytes using clinical HIV-1 isolates, and this represents an essential step toward clinical anti-HIV-1 gene therapy.

The Efficiency of Simian Foamy Virus Vector Type-1 (SFV-1) in Nondividing Cells and in Human PBLs

“…Simian foamy viruses have been readily isolated from PBLs of naturally and experimentally infected animals as well as infected animal caretakers (Falcone et al, 1999;Heneine et al, 1998;Hooks and Detrick-Hooks, 1981;Neuman-Haefelin et al, 1993;von Laer et al, 1996). To test the efficiency of SFV-1 vector in human PBLs, Ficoll gradient purified cells obtained from five different donors were independently PHA stimulated and infected twice with SFV-1 vector at a (Table 3), which is significantly higher than that of other retroviral vectors (Douglas et al, 1999;Vandendriessche et al, 1995;Yang et al, 1999). Transduction efficiency of human PBLs with the MuLV vector was Ͻ1%, again consistent with previous reports of infection of PBLs with equivalent moi (Culver et al, 1990(Culver et al, , 1991Morgan et al, 1994).…”

“…This low-level infection is attributed to MuLV's inefficiency at transducing nondividing cells and restricted replication in human PBLs despite entry (Naldini et al, 1996;Woffendin et al, 1994). A higher transduction efficiency ranging from 20 to 50% in human primary lymphocytes has been accomplished with MuLV vectors only after enhancing vector titers by pseudotyping with vesicular stomtitis virus envelope glycoprotein G (VSV-G) and improving the method of infections with complex manipulations of the system (Burns et al, 1993;Pollok et al, 1998;Sharma et al, 1996;Vandendriessche et al, 1995;Yang et al, 1999).…”

High-efficiency gene transfer into CD34+ cells with a human immunodeficiency virus type 1-based retroviral vector pseudotyped with vesicular stomatitis virus envelope glycoprotein G

“…One immediate application is to introduce anti-HIV-1 therapeutic genes (e.g., trans-dominant proteins, ribozymes, etc.) into CD34 ϩ cells (2,15,24,26,30). The transduced CD34 ϩ cells could be used to reconstitute SCID-hu mice to derive CD4 ϩ cells expressing these constructs.…”

Gene Therapy for Infectious Diseases

“…A Rev TNP that contains a single point mutation at leucine 78 was developed by our group. It inhibited HIV-1 replication in T-cell lines and PBL when challenged with both laboratory and clinical HIV-1 isolates (28,145). Another type of Rev TNP was generated by deletion of the nucleolar localization signal sequence (63).…”