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Title: Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike

Authors:
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  1. Weill-Med
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
INDUSTRY
OSTI Identifier:
1252760
Resource Type:
Journal Article
Resource Relation:
Journal Name: Nature Communications; Journal Volume: 6; Journal Issue: 09, 2015
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Lee, Jeong Hyun, Leaman, Daniel P., Kim, Arthur S., Torrents de la Peña, Alba, Sliepen, Kwinten, Yasmeen, Anila, Derking, Ronald, Ramos, Alejandra, de Taeye, Steven W., Ozorowski, Gabriel, Klein, Florian, Burton, Dennis R., Nussenzweig, Michel C., Poignard, Pascal, Moore, John P., Klasse, Per Johan, Sanders, Rogier W., Zwick, Michael B., Wilson, Ian A., Ward, Andrew B., Rockefeller), and Scripps). Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike. United States: N. p., 2016. Web. doi:10.1038/ncomms9167.
Lee, Jeong Hyun, Leaman, Daniel P., Kim, Arthur S., Torrents de la Peña, Alba, Sliepen, Kwinten, Yasmeen, Anila, Derking, Ronald, Ramos, Alejandra, de Taeye, Steven W., Ozorowski, Gabriel, Klein, Florian, Burton, Dennis R., Nussenzweig, Michel C., Poignard, Pascal, Moore, John P., Klasse, Per Johan, Sanders, Rogier W., Zwick, Michael B., Wilson, Ian A., Ward, Andrew B., Rockefeller), & Scripps). Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike. United States. doi:10.1038/ncomms9167.
Lee, Jeong Hyun, Leaman, Daniel P., Kim, Arthur S., Torrents de la Peña, Alba, Sliepen, Kwinten, Yasmeen, Anila, Derking, Ronald, Ramos, Alejandra, de Taeye, Steven W., Ozorowski, Gabriel, Klein, Florian, Burton, Dennis R., Nussenzweig, Michel C., Poignard, Pascal, Moore, John P., Klasse, Per Johan, Sanders, Rogier W., Zwick, Michael B., Wilson, Ian A., Ward, Andrew B., Rockefeller), and Scripps). 2016. "Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike". United States. doi:10.1038/ncomms9167.
@article{osti_1252760,
title = {Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike},
author = {Lee, Jeong Hyun and Leaman, Daniel P. and Kim, Arthur S. and Torrents de la Peña, Alba and Sliepen, Kwinten and Yasmeen, Anila and Derking, Ronald and Ramos, Alejandra and de Taeye, Steven W. and Ozorowski, Gabriel and Klein, Florian and Burton, Dennis R. and Nussenzweig, Michel C. and Poignard, Pascal and Moore, John P. and Klasse, Per Johan and Sanders, Rogier W. and Zwick, Michael B. and Wilson, Ian A. and Ward, Andrew B. and Rockefeller) and Scripps)},
abstractNote = {},
doi = {10.1038/ncomms9167},
journal = {Nature Communications},
number = 09, 2015,
volume = 6,
place = {United States},
year = 2016,
month = 7
}
  • Following CD4 receptor binding to the HIV-1 envelope spike (Env), the conserved N-heptad repeat (NHR) region of gp41 forms a coiled-coil that is a precursor to the fusion reaction. Although it has been a target of drug and vaccine design, there are few monoclonal antibody (mAb) tools with which to probe the antigenicity and immunogenicity specifically of the NHR coiled-coil. Here, we have rescued HIV-1-neutralizing anti-NHR mAbs from immune phage display libraries that were prepared (i) from b9 rabbits immunized with a previously described mimetic of the NHR coiled-coil, N35{sub CCG}-N13, and (ii) from an HIV-1 infected individual. We describemore » a rabbit single-chain Fv fragment (scFv), 8K8, and a human Fab, DN9, which specifically recognize NHR coiled-coils that are unoccupied by peptide corresponding to the C-heptad repeat or CHR region of gp41 (e.g. C34). The epitopes of 8K8 and DN9 were found to partially overlap with that of a previously described anti-NHR mAb, IgG D5; however, 8K8 and DN9 were much more specific than D5 for unoccupied NHR trimers. The mAbs, including a whole IgG 8K8 molecule, neutralized primary HIV-1 of clades B and C in a pseudotyped virus assay with comparable, albeit relatively modest potency. Finally, a human Fab T3 and a rabbit serum (both non-neutralizing) were able to block binding of D5 and 8K8 to a gp41 NHR mimetic, respectively, but not the neutralizing activity of these mAbs. We conclude from these results that NHR coiled-coil analogs of HIV-1 gp41 elicit many Abs during natural infection and through immunization, but that due to limited accessibility to the corresponding region on fusogenic gp41 few can neutralize. Caution is therefore required in targeting the NHR for vaccine design. Nevertheless, the mAb panel may be useful as tools for elucidating access restrictions to the NHR of gp41 and in designing potential improvements to mimetics of receptor-activated Env.« less
  • Cited by 11
  • The C-terminal alpha-helix of gp41 membrane-proximal external region (MPER; {sup 671}NWFDITNWLWYIK{sup 683}) encompassing 4E10/10E8 epitopes is an attractive target for HIV-1 vaccine development. We previously reported that gp41-HR1-54Q, a trimeric protein comprised of the MPER in the context of a stable six-helix bundle (6HB), induced strong immune responses against the helix, but antibodies were directed primarily against the non-neutralizing face of the helix. To better target 4E10/10E8 epitopes, we generated four putative fusion intermediates by introducing double point mutations or deletions in the heptad repeat region 1 (HR1) that destabilize 6HB in varying degrees. One variant, HR1-∆10-54K, elicited antibodies inmore » rabbits that targeted W672, I675 and L679, which are critical for 4E10/10E8 recognition. Overall, the results demonstrated that altering structural parameters of 6HB can influence immunogenic properties of the MPER and antibody targeting. Further exploration of this strategy could allow development of immunogens that could lead to induction of 4E10/10E8-like antibodies. - Highlights: • Four gp41 MPER-based immunogens that resemble fusion intermediates were generated. • C-terminal region of MPER that contains 4E10/10E8 epitopes was highly immunogenic. • Altering 6HB structure can influence immunogenic properties of the MPER. • Induced antibodies targeted multiple residues critical for 4E10/10E8 binding. • Development of immunogens based on fusion intermediates is a promising strategy.« less
  • Asymmetric disposition of Fab arms in the structures solved for the broadly neutralizing monoclonal antibody (nmAb) IgG1 b12 raised the question of whether the unusual shape observed for b12 is common for all IgG1 mAbs or if there is a difference in the overall shape of nmAbs versus non-nmAbs. In this paper, we compared small angle x-ray scattering (SAXS) data-based models and limited proteolysis profiles of some IgG1 mAbs known to be having and lacking HIV-1 neutralizing potency. In non-nmAbs, the Fab arms were found to be symmetrically disposed in space relative to central Fc, but in most nmAbs, themore » Fab arms were asymmetrically disposed, as seen for IgG1 b12. The only exceptions were 2G12 and 4E10, where both Fab arms were closed above Fc, suggesting some Fab-Fc and/or Fab-Fab interaction in the nmAbs that constrained extension of the Fab-Fc linker. Interestingly, these observations were correlated with differential proteolysis profiles of the mAbs by papain. Under conditions when papain could cut both Fab arms of non-nmAbs, only one Fab arm could be removed from neutralizing ones (except for 2G12 and 4E10). Chromatography and small angle x-ray scattering results of papain-digested products revealed that 1) the Fab-Fc or Fab-Fab interactions in unliganded mAbs are retained in digested products, and 2) whereas anti-gp120 non-nmAbs could bind two gp120 molecules, nmAbs could bind only one gp120. Finally, additional experiments showed that except for 2G12 and 4E10, unopen shapes of nmAbs remain uninfluenced by ionic strength but can be reversibly opened by low pH of buffer accompanied by loss of ligand binding ability.« less
  • Antibody Z13e1 is a relatively broadly neutralizing anti-human immunodeficiency virus type 1 antibody that recognizes the membrane-proximal external region (MPER) of the human immunodeficiency virus type 1 envelope glycoprotein gp41. Based on the crystal structure of an MPER epitope peptide in complex with Z13e1 Fab, we identified an unrelated protein, interleukin (IL)-22, with a surface-exposed region that is structurally homologous in its backbone to the gp41 Z13e1 epitope. By grafting the gp41 Z13e1 epitope sequence onto the structurally homologous region in IL-22, we engineered a novel protein (Z13-IL22-2) that contains the MPER epitope sequence for use as a potential immunogenmore » and as a reagent for the detection of Z13e1-like antibodies. The Z13-IL22-2 protein binds Fab Z13e1 with a K{sub d} of 73 nM. The crystal structure of Z13-IL22-2 in complex with Fab Z13e1 shows that the epitope region is faithfully replicated in the Fab-bound scaffold protein; however, isothermal calorimetry studies indicate that Fab binding to Z13-IL22-2 is not a lock-and-key event, leaving open the question of whether conformational changes upon binding occur in the Fab, in Z13-IL-22, or in both.« less