title: Analysis of the HIV-1 gp41 specific immune response using a multiplexed antibody detection assay.
abstract: A fluorescence-based, multiplexed, antibody-binding and mapping assay was developed to characterize antibody responses in HIV-1-infected individuals to the ectodomain of the HIV-1 gp41 envelope glycoprotein. The antigen panel included intact recombinant gp41, the fusion peptide region, the polar region, the N-heptad region, the C-heptad region as well as overlapping epitopes in the 2F5 and 4E10 monoclonal antibody-binding regions. The panel included both native and constrained peptides specifically designed to mimic putative gp41 prefusion and fusion intermediates. The results of these analyses revealed a broad pattern of immune responses against the test antigens, suggesting that none of these gp41 regions are immunologically silent. The HIV-1-positive sera were also evaluated using infectivity inhibition assays. No correlation was evident between the breadth or magnitude of specific anti-gp41 reactivities and virus neutralization potency. These evaluations demonstrated the substantial potential of the multiplexed antibody binding and mapping assay for rapid and sensitive analysis of complex antibody responses.
authors.affiliation: Vaccine and Biologics Research, Merck Research Laboratories, WYN-2, MRL-Wayne, 466 Devon Park Drive, Wayne, PA 19087-8630, USA.
authors.forename: David
authors.initials: D
authors.lastname: Opalka
authors.forename: Antonello
authors.initials: A
authors.lastname: Pessi
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Bianchi
authors.forename: Gennaro
authors.initials: G
authors.lastname: Ciliberto
authors.forename: William
authors.initials: W
authors.lastname: Schleif
authors.forename: Michael
authors.initials: M
authors.lastname: McElhaugh
authors.forename: Renee
authors.initials: R
authors.lastname: Danzeisen
authors.forename: Romas
authors.initials: R
authors.lastname: Geleziunas
authors.forename: Michael
authors.initials: M
authors.lastname: Miller
authors.forename: Debra M
authors.initials: DM
authors.lastname: Eckert
authors.forename: David
authors.initials: D
authors.lastname: Bramhill
authors.forename: Joseph
authors.initials: J
authors.lastname: Joyce
authors.forename: James
authors.initials: J
authors.lastname: Cook
authors.forename: William
authors.initials: W
authors.lastname: Magilton
authors.forename: John
authors.initials: J
authors.lastname: Shiver
authors.forename: Emilio
authors.initials: E
authors.lastname: Emini
authors.forename: Mark T
authors.initials: MT
authors.lastname: Esser
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods*; D015483:HIV Antibodies / Q000032:analysis*; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000276:immunology*; D015658:HIV Infections / Q000097:blood / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011487:Protein Conformation; D012680:Sensitivity and Specificity
chemical_list: D015483:HIV Antibodies; D015700:HIV Envelope Protein gp41