Potent inhibition of drug-resistant HIV protease variants by monoclonal antibodies.
The monoclonal antibodies 1696 and F11.2.32 strongly inhibit the activity of wild-type HIV-1 protease (PR) by binding to epitopes at the enzyme N-terminus (residues 1-6) and flap residues 36-46, respectively. Here we demonstrate that these antibodies are also potent inhibitors of PR variants resistant to active-site inhibitors used as anti-AIDS drugs. Our in vitro experiments revealed that the inhibitory potency of single-chain fragments (scFv) of these antibodies is not significantly affected by the presence of mutations in PR; inhibition constants for drug-resistant protease variants are 5-11 nM and 13-169 nM for scFv1696 and for scFvF11.2.32, respectively. Tethered dimer of HIV-1 PR variant proved to be a model protease variant resistant to dissociative inhibition by 1696, and, strikingly, it also displayed resistance to inhibition by F11.2.32 suggesting that dimer dissociation also plays a role in the inhibitory action of F11.2.32.
Bartonová
Vanda
V
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nam. 2, 166 37 Prague 6, Czech Republic.
Král
Vlastimil
V
Sieglová
Irena
I
Brynda
Jirí
J
Fábry
Milan
M
Horejsí
Magdalena
M
Kozísek
Milan
M
Sasková
Klára Grantz
KG
Konvalinka
Jan
J
Sedlácek
Juraj
J
Rezácová
Pavlína
P
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D023241:Antiretroviral Therapy, Highly Active; D019281:Dimerization; D024882:Drug Resistance, Viral / Q000235:genetics*; D014644:Genetic Variation; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology; D016333:HIV Protease / Q000187:drug effects* / Q000235:genetics / Q000276:immunology; D017320:HIV Protease Inhibitors / Q000494:pharmacology; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology / Q000494:pharmacology*; D008958:Models, Molecular; D009154:Mutation; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D017320:HIV Protease Inhibitors; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D016333:HIV Protease