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

Abstract

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.

• Authors: Cinque Soto, Gilad Ofek, M Gordon Joyce, Baoshan Zhang, Krisha McKee, Nancy S Longo, Yongping Yang, Jinghe Huang, Robert Parks, Joshua Eudailey, Krissey E Lloyd, S Munir Alam, Barton F Haynes, NISC Comparative Sequencing Program, James C Mullikin, Mark Connors, John R Mascola, Lawrence Shapiro, Peter D Kwong
• Journal: PloS one (PLoS One) Vol. 11 Issue 6 Pg. e0157409 ( 2016) ISSN: 1932-6203 [Electronic] United States
• PMID: 27299673 (Publication Type: Journal Article)

Chemical References

• Antibodies, Neutralizing
• Complementarity Determining Regions
• HIV Antibodies
• HIV Envelope Protein gp41

Topics

• Amino Acid Sequence
• Antibodies, Neutralizing (chemistry, genetics, immunology)
• Cell Line
• Complementarity Determining Regions (chemistry, genetics, immunology)
• Crystallography, X-Ray
• HIV Antibodies (chemistry, genetics, immunology)
• HIV Envelope Protein gp41 (chemistry, immunology)
• HIV Infections (genetics, immunology)
• HIV-1 (chemistry, immunology)
• Humans
• Models, Molecular
• Mutagenesis