title: Identification and characterization of an immunogenic hybrid epitope formed by both HIV gp120 and human CD4 proteins.
abstract: Certain antibodies from HIV-infected humans bind conserved transition state (CD4 induced [CD4i]) domains on the HIV envelope glycoprotein, gp120, and demonstrate extreme dependence on the formation of a gp120-human CD4 receptor complex. The epitopes recognized by these antibodies remain undefined although recent crystallographic studies of the anti-CD4i monoclonal antibody (MAb) 21c suggest that contacts with CD4 as well as gp120 might occur. Here, we explore the possibility of hybrid epitopes that demand the collaboration of both gp120 and CD4 residues to enable antibody reactivity. Analyses with a panel of human anti-CD4i MAbs and gp120-CD4 antigens with specific mutations in predicted binding domains revealed one putative hybrid epitope, defined by the human anti-CD4i MAb 19e. In virological and immunological tests, MAb 19e did not bind native or constrained gp120 except in the presence of CD4. This contrasted with other anti-CD4i MAbs, including MAb 21c, which bound unliganded, full-length gp120 held in a constrained conformation. Conversely, MAb 19e exhibited no specific reactivity with free human CD4. Computational modeling of MAb 19e interactions with gp120-CD4 complexes suggested a distinct binding profile involving antibody heavy chain interactions with CD4 and light chain interactions with gp120. In accordance, targeted mutations in CD4 based on this model specifically reduced MAb 19e interactions with stable gp120-CD4 complexes that retained reactivity with other anti-CD4i MAbs. These data represent a rare instance of an antibody response that is specific to a pathogen-host cell protein interaction and underscore the diversity of immunogenic CD4i epitope structures that exist during natural infection.
authors: Institute of Human Virology, Department of Medicine, University of Maryland School of Medicine, University of Maryland, Baltimore, 725 West Lombard Street, Baltimore, MD 21201, USA.
authors: George K
authors: GK
authors: Lewis
authors: Timothy R
authors: TR
authors: Fouts
authors: Sani
authors: S
authors: Ibrahim
authors: Brian M
authors: BM
authors: Taylor
authors: Rachita
authors: R
authors: Salkar
authors: Yongjun
authors: Y
authors: Guan
authors: Roberta
authors: R
authors: Kamin-Lewis
authors: James E
authors: JE
authors: Robinson
authors: Anthony L
authors: AL
authors: Devico
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D015704:CD4 Antigens / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D015483:HIV Antibodies / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D006801:Humans; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D011485:Protein Binding
chemical_list: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120