Development and characterization of ten monoclonal anti-Vpr antibodies.
HIV-1 Vpr is a 96-amino acid auxiliary protein that performs numerous activities during viral infection. In the present study, 10 antibodies were generated after mice immunization with either the N- or the C-terminus domain of Vpr, respectively, Vpr(1-51) and Vpr(52-96). ELISA and immunoblot experiments using pure synthetic overlapping Vpr peptides suggested that these anti-Vpr antibodies could be classified into five groups and that they recognized conformational or linear Vpr epitopes. Further analysis revealed the effect of C-terminal arginine mutations on the antibody binding. Two of the antibodies precipitated Vpr expressed after transfection of a Vpr-encoding vector in human cells. More importantly, one of them was able to detect Vpr in HIV-1-infected U1 cells and in HIV-1-infected human PBMC. Surface plasmon resonance experiments demonstrated that some of these antibodies prevented the interaction between Vpr and one of its cellular partners, the adenine nucleotide translocator. Thus, these anti-Vpr monoclonal antibodies may be useful to any laboratory working on the molecular mechanism of HIV-1 infection.
Sabbah
Emmanuelle N
EN
Unité de Pharmacochimie Moléculaire et Structurale, INSERM U266, CNRS 8600, Université René Descartes, UFR des Sciences Pharmaceutiques et Biologiques, 75006 Paris, France.
Delaunay
Thierry
T
Varin
Audrey
A
Le-Rouzic
Erwann
E
Benichou
Serge
S
Herbein
Georges
G
Druillennec
Sabine
S
Roques
Bernard P
BP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D018604:Epitope Mapping / Q000379:methods; D005260:Female; D016336:Gene Products, vpr / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D007963:Leukocytes, Mononuclear / Q000821:virology*; D051379:Mice; D008807:Mice, Inbred BALB C; D020349:Surface Plasmon Resonance; D054325:vpr Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; D016336:Gene Products, vpr; D054325:vpr Gene Products, Human Immunodeficiency Virus