Intracellular immunization against HIV-1 infection of human T lymphocytes: utility of anti-rev single-chain variable fragments.
Genetic therapy offers a potentially promising approach with which to combat human immunodeficiency virus type 1 (HIV-1) infections. Several modalities, using protein- and RNA-based systems, have recently been shown to inhibit HIV-1 replication. A single-chain variable fragment (SFv), constructed from the cDNA of a monoclonal antibody to the HIV-1 regulatory protein Rev, has been demonstrated to potently inhibit HIV-1 replication, when expressed intracellularly in an epithelial cell-line (HeLa-CD4). Murine retroviral shuttle vectors, which express the anti-Rev SFv moiety, have now been constructed. HIV-1 infection was dramatically inhibited in human T-lymphocytic cell-lines, CEM and Sup-T1, transduced with these anti-Rev SFv-expressing vectors. This resistance to high levels of HIV-1 expression was demonstrated in both mixed populations and clones of these cells. Of further potential clinical significance, HIV-1 infection was also potently inhibited in human peripheral blood mononuclear cells (PBMC), transduced with retroviral vectors expressing the anti-Rev SFv molecule. These data suggest that intracellular expression of anti-Rev SFvs, or related approaches, may be utilized as genetic therapy, or intracellular immunization, for HIV-1 infections in vivo.
Duan
L
L
Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Zhu
M
M
Bagasra
O
O
Pomerantz
R J
RJ
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D017931:DNA Primers; D015870:Gene Expression; D015694:Gene Products, rev; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D015497:HIV-1 / Q000276:immunology / Q000502:physiology*; D006801:Humans; D007135:Immunoglobulin Variable Region; D009052:Leukemia Virus, Murine / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000821:virology; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D013601:T-Lymphocytes / Q000821:virology*; D014162:Transfection; D016662:Virus Integration; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D015694:Gene Products, rev; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C092667:anti-HIV-1 Rev SFv; D054321:rev Gene Products, Human Immunodeficiency Virus