Developmental Pathway of the MPER-Directed HIV-1-Neutralizing Antibody 10E8

Journal Article · Tue Jun 14 00:00:00 EDT 2016 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0157409· OSTI ID:1258680

Soto, Cinque ^[1]; Ofek, Gilad ^[1]; Joyce, M. Gordon ^[1]; Zhang, Baoshan ^[1]; McKee, Krisha ^[1]; Longo, Nancy S. ^[1]; Yang, Yongping ^[1]; Huang, Jinghe ^[1]; Parks, Robert ^[2]; Eudailey, Joshua ^[2]; Lloyd, Krissey E. ^[2]; Alam, S. Munir ^[2]; Haynes, Barton F. ^[2]; Mullikin, James C. ^[1]; Connors, Mark ^[1]; Mascola, John R. ^[1]; Shapiro, Lawrence ^[3]; Kwong, Peter D. ^[1]; Jiang, Shibo ^[4]

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1. National Inst. of Health (NIH), Bethesda, MD (United States)
2. Duke Univ. Medical Center, Durham, NC (United States)
3. National Inst. of Health (NIH), Bethesda, MD (United States); Columbia Univ., New York, NY (United States)
4. Fudan Univ. (China)

Antibody 10E8 targets the membrane-proximal external region (MPER) of HIV-1 gp41, neutralizes >97% of HIV-1 isolates, and lacks the auto-reactivity often associated with MPER-directed antibodies. The developmental pathway of 10E8 might therefore serve as a promising template for vaccine design, but samples from time-of-infection—often used to infer the B cell record—are unavailable. In this study, we used crystallography, next-generation sequencing (NGS), and functional assessments to infer the 10E8 developmental pathway from a single time point. Mutational analysis indicated somatic hypermutation of the 2nd-heavy chain-complementarity determining region (CDR H2) to be critical for neutralization, and structures of 10E8 variants with V-gene regions reverted to genomic origin for heavy-and-light chains or heavy chain-only showed structural differences >2 Å relative to mature 10E8 in the CDR H2 and H3. To understand these developmental changes, we used bioinformatic sieving, maximum likelihood, and parsimony analyses of immunoglobulin transcripts to identify 10E8-lineage members, to infer the 10E8-unmutated common ancestor (UCA), and to calculate 10E8-developmental intermediates. We were assisted in this analysis by the preservation of a critical D-gene segment, which was unmutated in most 10E8-lineage sequences. UCA and early intermediates weakly bound a 26-residue-MPER peptide, whereas HIV-1 neutralization and epitope recognition in liposomes were only observed with late intermediates. Antibody 10E8 thus develops from a UCA with weak MPER affinity and substantial differences in CDR H2 and H3 from the mature 10E8; only after extensive somatic hypermutation do 10E8-lineage members gain recognition in the context of membrane and HIV-1 neutralization.

• Research Organization: Argonne National Laboratory (ANL), Argonne, IL (United States)
• Sponsoring Organization: Intramural Research Program of the Vaccine Research Center; National Inst. of Allergy and Infectious Diseases; National Human Genome Research Inst.; National Inst. of Health; Center for HIV/AIDS Vaccine Immunology-Immunogen Design
• Contributing Organization: NISC Comparative Sequencing Program
• Grant/Contract Number: UM1 AI100645
• OSTI ID: 1258680
• Journal Information: PLoS ONE, Vol. 11, Issue 6; ISSN 1932-6203
• Publisher: Public Library of Science
• Country of Publication: United States
• Language: ENGLISH

Related Subjects

60 APPLIED LIFE SCIENCES
Phylogenetic analysis
Sequence motif analysis
Sequence alignment
Antibodies
Multiple alignment calculation
HIV-1
Enzyme-linked immunoassays
Sequence analysis