Abstract
A variety of genetic therapies or intracellular immunization techniques hold promise as modalities to inhibit human immunodeficiency virus type 1 (HIV-1) replication in vivo. We have recently demonstrated that a single-chain variable fragment (SFv) construct, derived from a monoclonal antibody that binds to the HIV-1 regulatory protein Rev, can be expressed intracellularly and potently inhibits HIV-1 replication. This single-chain intracellular antibody, which avidly binds to the effector domain of Rev, is now demonstrated to dramatically inhibit various diverse laboratory and primary clinical strains of HIV-1. Potent suppression of HIV-1 replication by this modality is maintained over several months in long-term cultures. As well, the intracellular expression of anti-Rev SFv is shown to alter HIV-1 replication by specifically affecting Rev function. Importantly, no alterations in HIV-1 internalization, reverse transcription, or initial transcription of multiply spliced viral mRNAs are demonstrated in SFv-immunized cells, as compared to controls. Thus, these studies extend the understanding of the molecular mechanisms involved in the inhibition of lentivirus replication, by these intracellular antibody constructs.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Antibodies, Monoclonal / chemistry
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / immunology*
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Antiviral Agents / pharmacology*
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Base Sequence
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DNA, Viral / isolation & purification
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Gene Expression Regulation, Viral / drug effects
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Gene Products, rev / antagonists & inhibitors*
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Gene Products, rev / immunology
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Gene Products, rev / physiology
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HIV Antibodies / chemistry
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HIV Antibodies / genetics
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HIV Antibodies / immunology*
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HIV-1 / drug effects*
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HIV-1 / immunology
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HIV-1 / physiology
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HeLa Cells
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Humans
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Immunization, Passive / methods*
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Immunoglobulin Fragments / genetics
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Immunoglobulin Fragments / pharmacology*
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Immunoglobulin Variable Region / genetics
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Immunoglobulin Variable Region / pharmacology*
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Molecular Sequence Data
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Polymerase Chain Reaction
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Proviruses / isolation & purification
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / pharmacology*
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Recombinant Proteins
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Single-Chain Antibodies
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Transfection
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Virus Replication / drug effects*
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rev Gene Products, Human Immunodeficiency Virus
Substances
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Antibodies, Monoclonal
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Antiviral Agents
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DNA, Viral
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Gene Products, rev
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HIV Antibodies
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Immunoglobulin Fragments
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Immunoglobulin Variable Region
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Recombinant Fusion Proteins
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Recombinant Proteins
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Single-Chain Antibodies
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anti-HIV-1 Rev SFv
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rev Gene Products, Human Immunodeficiency Virus