Molecular and virological effects of intracellular anti-Rev single-chain variable fragments on the expression of various human immunodeficiency virus-1 strains.
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.
Duan
L
L
Dorrance H. Hamilton Laboratories, Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107.
Zhang
H
H
Oakes
J W
JW
Bagasra
O
O
Pomerantz
R J
RJ
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D004279:DNA, Viral / Q000302:isolation & purification; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D015694:Gene Products, rev / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000502:physiology; D015483:HIV Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015497:HIV-1 / Q000187:drug effects* / Q000276:immunology / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D007116:Immunization, Passive / Q000379:methods*; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000494:pharmacology*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000494:pharmacology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects*; D054321:rev Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D004279:DNA, Viral; D015694:Gene Products, rev; D015483:HIV Antibodies; D007128:Immunoglobulin Fragments; 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