title: Enhancement of alpha -helicity in the HIV-1 inhibitory peptide DP178 leads to an increased affinity for human monoclonal antibody 2F5 but does not elicit neutralizing responses in vitro. Implications for vaccine design.
abstract: The synthetic peptide DP178, derived from the carboxyl-terminal heptad repeat region of human immunodeficiency virus type 1 GP41 protein is a potent inhibitor of viral-mediated fusion and contains the sequence ELDKWA, which constitutes the recognition epitope for the broadly neutralizing human monoclonal antibody 2F5. Efforts at eliciting a 2F5-like immune response by immunization with peptides or fusion proteins containing this sequence have not met with success, possibly because of incorrect structural presentation of the epitope. Although the structure of the carboxyl-terminal heptad repeat on the virion is not known, several recent reports have suggested a propensity for alpha-helical conformation. We have examined DP178 in the context of a model for optimized alpha-helices and show that the native sequence conforms poorly to the model. Solution conformation of DP178 was studied by circular dichroism and NMR spectroscopy and found to be predominantly random, consistent with previous reports. NMR mapping was used to show that the low percentage of alpha-helix present was localized to residues Glu(662) through Asn(671), a region encompassing the 2F5 epitope. Using NH(2)-terminal extensions derived from either GP41 or the yeast GCN4 leucine zipper dimerization domain, we designed peptide analogs in which the average helicity is significantly increased compared with DP178 and show that these peptides exhibit both a modest increase in affinity for 2F5 using a novel competitive solution-based binding assay and an increased ability to inhibit viral entry in a single-cycle infectivity model. Selected peptides were conjugated to carrier protein and used for guinea pig immunizations. High peptide-specific titers were achieved using these immunogens, but the resulting sera were incapable of viral neutralization. We discuss these findings in terms of structural and immunological considerations as to the utility of a 2F5-like response.
authors.affiliation: Department of Virus and Cell Biology, Merck Research Laboratories, West Point, Pennsylvania 19486, USA. joseph_joyce@merck.com
authors.forename: Joseph G
authors.initials: JG
authors.lastname: Joyce
authors.forename: William M
authors.initials: WM
authors.lastname: Hurni
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Bogusky
authors.forename: Victor M
authors.initials: VM
authors.lastname: Garsky
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Liang
authors.forename: Michael P
authors.initials: MP
authors.lastname: Citron
authors.forename: Renee C
authors.initials: RC
authors.lastname: Danzeisen
authors.forename: Michael D
authors.initials: MD
authors.lastname: Miller
authors.forename: John W
authors.initials: JW
authors.lastname: Shiver
authors.forename: Paul M
authors.initials: PM
authors.lastname: Keller
mesh_terms: D016915:AIDS Vaccines / Q000737:chemistry / Q000276:immunology*; D000595:Amino Acid Sequence; D019380:Anti-HIV Agents / Q000737:chemistry / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002942:Circular Dichroism; D000077560:Enfuvirtide; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*
chemical_list: D016915:AIDS Vaccines; D019380:Anti-HIV Agents; D000911:Antibodies, Monoclonal; D015700:HIV Envelope Protein gp41; D010446:Peptide Fragments; D000077560:Enfuvirtide