Targeting human immunodeficiency virus type 1 reverse transcriptase by intracellular expression of single-chain variable fragments to inhibit early stages of the viral life cycle.
Novel molecular approaches to inhibit human immunodeficiency virus type 1 (HIV-1) infection have received increasing attention because of the lack of effective antiviral drug therapies in vivo. We now demonstrate that cells can be intracellularly immunized by cytoplasmic expression of single-chain variable antibody fragments (SFv) which bind to the HIV-1 reverse transcriptase (RT) enzyme. The expression of anti-RT SFv in T-lymphocytic cells specifically neutralizes the RT activity in the preintegration stage and affects the reverse transcription process, an early event of the HIV-1 life cycle. Blocking the virus at these early stages dramatically decreased HIV-1 propagation, as well as the HIV-1-induced cytopathic effects in susceptible human T lymphocytes, by impeding the formation of the proviral DNA. These data also demonstrate that intracellular, complete SFvs may gain access to viral proteins of the HIV-1 preintegration complex. These SFvs will provide a tool with which to better understand the molecular mechanisms involved in restricting viral replication in HIV-1-infected cells.
Shaheen
F
F
Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Duan
L
L
Zhu
M
M
Bagasra
O
O
Pomerantz
R J
RJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics*; D000998:Antiviral Agents / Q000494:pharmacology*; D003001:Cloning, Molecular; D054303:HIV Reverse Transcriptase; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D012194:RNA-Directed DNA Polymerase / Q000276:immunology*; D011994:Recombinant Proteins / Q000494:pharmacology; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology*; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; C055966:immunoglobulin Fv; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase