Identification of a linear peptide recognized by monoclonal antibody 2D7 capable of generating CCR5-specific antibodies with human immunodeficiency virus-neutralizing activity.
CCR5 is the major coreceptor for human immunodeficiency virus (HIV) infection. The murine monoclonal antibody (MAb) 2D7, which recognizes a conformation-dependent epitope in the second extracellular loop of CCR5, is one of the most potent inhibitors of R5 virus cell entry. However, attempts to humanize 2D7 for in vivo human use have been unsuccessful so far. A filamentous phage library expressing random peptides was used to identify a peptide mimitope that is recognized by MAb 2D7. A synthetic peptide containing this sequence (2D7-2SK) bound to MAb 2D7 with high affinity and reversed its HIV type 1 (HIV-1) fusion inhibitory activity. The peptide contains sequence homologies to two distal regions of the second extracellular loop of human CCR5, both of which are required for MAb 2D7 binding. Rabbit anti-2D7-mimitope antibodies competed with MAb 2D7 for binding to the 2D7-2SK peptide in Biacore biosensor testing. Importantly, the rabbit anti-2D7-2SK antibodies bound to CCR5 on cells and specifically inhibited R5 (but not X4) envelope-mediated syncytium formation. These antibodies also neutralized infection of human peripheral blood mononuclear cells with R5 HIV isolates comparably to MAb 2D7. In summary, we have identified a novel peptide that closely mimics the MAb 2D7 epitope on CCR5. This peptide could be included as a potential vaccine candidate or to isolate 2D7-like human antibodies as entry inhibitors for R5 viruses.
Khurana
Surender
S
Division of Viral Products, Center for Biologics Evaluation and Research, FDA, Bethesda, MD 20892, USA.
Kennedy
Michael
M
King
Lisa R
LR
Golding
Hana
H
D016915:AIDS Vaccines / Q000235:genetics / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D065100:CCR5 Receptor Antagonists; D002460:Cell Line; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D015483:HIV Antibodies; D015497:HIV-1 / Q000276:immunology* / Q000472:pathogenicity; D006801:Humans; D066298:In Vitro Techniques; D008253:Macaca mulatta; D008561:Membrane Fusion; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D011817:Rabbits; D019713:Receptors, CCR5 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D016915:AIDS Vaccines; D000911:Antibodies, Monoclonal; D065100:CCR5 Receptor Antagonists; D000939:Epitopes; D015483:HIV Antibodies; D010446:Peptide Fragments; D019713:Receptors, CCR5