Analysis of the HIV-1 gp41 specific immune response using a multiplexed antibody detection assay.
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.
Opalka
David
D
Vaccine and Biologics Research, Merck Research Laboratories, WYN-2, MRL-Wayne, 466 Devon Park Drive, Wayne, PA 19087-8630, USA.
Pessi
Antonello
A
Bianchi
Elisabetta
E
Ciliberto
Gennaro
G
Schleif
William
W
McElhaugh
Michael
M
Danzeisen
Renee
R
Geleziunas
Romas
R
Miller
Michael
M
Eckert
Debra M
DM
Bramhill
David
D
Joyce
Joseph
J
Cook
James
J
Magilton
William
W
Shiver
John
J
Emini
Emilio
E
Esser
Mark T
MT
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
D015483:HIV Antibodies; D015700:HIV Envelope Protein gp41