title: Antibody elicited against the gp41 N-heptad repeat (NHR) coiled-coil can neutralize HIV-1 with modest potency but non-neutralizing antibodies also bind to NHR mimetics.
abstract: Following CD4 receptor binding to the HIV-1 envelope spike (Env), the conserved N-heptad repeat (NHR) region of gp41 forms a coiled-coil that is a precursor to the fusion reaction. Although it has been a target of drug and vaccine design, there are few monoclonal antibody (mAb) tools with which to probe the antigenicity and immunogenicity specifically of the NHR coiled-coil. Here, we have rescued HIV-1-neutralizing anti-NHR mAbs from immune phage display libraries that were prepared (i) from b9 rabbits immunized with a previously described mimetic of the NHR coiled-coil, N35(CCG)-N13, and (ii) from an HIV-1 infected individual. We describe a rabbit single-chain Fv fragment (scFv), 8K8, and a human Fab, DN9, which specifically recognize NHR coiled-coils that are unoccupied by peptide corresponding to the C-heptad repeat or CHR region of gp41 (e.g. C34). The epitopes of 8K8 and DN9 were found to partially overlap with that of a previously described anti-NHR mAb, IgG D5; however, 8K8 and DN9 were much more specific than D5 for unoccupied NHR trimers. The mAbs, including a whole IgG 8K8 molecule, neutralized primary HIV-1 of clades B and C in a pseudotyped virus assay with comparable, albeit relatively modest potency. Finally, a human Fab T3 and a rabbit serum (both non-neutralizing) were able to block binding of D5 and 8K8 to a gp41 NHR mimetic, respectively, but not the neutralizing activity of these mAbs. We conclude from these results that NHR coiled-coil analogs of HIV-1 gp41 elicit many Abs during natural infection and through immunization, but that due to limited accessibility to the corresponding region on fusogenic gp41 few can neutralize. Caution is therefore required in targeting the NHR for vaccine design. Nevertheless, the mAb panel may be useful as tools for elucidating access restrictions to the NHR of gp41 and in designing potential improvements to mimetics of receptor-activated Env.
authors.affiliation: Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Josh D
authors.initials: JD
authors.lastname: Nelson
authors.forename: Heather
authors.initials: H
authors.lastname: Kinkead
authors.forename: Florence M
authors.initials: FM
authors.lastname: Brunel
authors.forename: Dan
authors.initials: D
authors.lastname: Leaman
authors.forename: Richard
authors.initials: R
authors.lastname: Jensen
authors.forename: John M
authors.initials: JM
authors.lastname: Louis
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Maruyama
authors.forename: Carole A
authors.initials: CA
authors.lastname: Bewley
authors.forename: Katherine
authors.initials: K
authors.lastname: Bowdish
authors.forename: G Marius
authors.initials: GM
authors.lastname: Clore
authors.forename: Philip E
authors.initials: PE
authors.lastname: Dawson
authors.forename: Shana
authors.initials: S
authors.lastname: Frederickson
authors.forename: Rose G
authors.initials: RG
authors.lastname: Mage
authors.forename: Douglas D
authors.initials: DD
authors.lastname: Richman
authors.forename: Dennis R
authors.initials: DR
authors.lastname: Burton
authors.forename: Michael B
authors.initials: MB
authors.lastname: Zwick
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics; D001665:Binding Sites; D002308:Cardiolipins / Q000276:immunology; D003429:Cross Reactions; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D015483:HIV Antibodies / Q000235:genetics; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary; D011817:Rabbits; D020449:Repetitive Sequences, Amino Acid; D017386:Sequence Homology, Amino Acid
chemical_list: D000911:Antibodies, Monoclonal; D002308:Cardiolipins; D000939:Epitopes; D015483:HIV Antibodies; D015700:HIV Envelope Protein gp41; C514787:gp41 protein, Human immunodeficiency virus 1