1. Analysis of the HIV-1 gp41 specific immune response using a multiplexed antibody detection assay
- Author
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Emilio A. Emini, Elisabetta Bianchi, David Bramhill, William Magilton, Gennaro Ciliberto, James C. Cook, Michael W. Miller, Mark T. Esser, John W. Shiver, Antonello Pessi, Joseph G. Joyce, William A. Schleif, Renee Danzeisen, David Opalka, Michael McElhaugh, Debra M Eckert, and Romas Geleziunas
- Subjects
Protein Conformation ,viruses ,Immunology ,Molecular Sequence Data ,Fluorescent Antibody Technique ,Enzyme-Linked Immunosorbent Assay ,HIV Infections ,Biology ,HIV Antibodies ,Gp41 ,Sensitivity and Specificity ,Epitope ,Antigen ,Neutralization Tests ,Immunology and Allergy ,Animals ,Humans ,Amino Acid Sequence ,Neutralizing antibody ,virus diseases ,Acquired immune system ,Flow Cytometry ,Virology ,Molecular biology ,HIV Envelope Protein gp41 ,Epitope mapping ,Ectodomain ,biology.protein ,HIV-1 ,Antibody ,Epitope Mapping - 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.
- Published
- 2003