Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.

To date, no immunization of humans or animals has elicited broadly neutralizing sera able to prevent HIV-1 transmission; however, elicitation of broad and potent heavy chain only antibodies (HCAb) has previously been reported in llamas. In this study, the anti-HIV immune responses in immunized llama...

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Main Authors: Laura E McCoy, Lucy Rutten, Dan Frampton, Ian Anderson, Luke Granger, Rachael Bashford-Rogers, Gillian Dekkers, Nika M Strokappe, Michael S Seaman, Willie Koh, Vanina Grippo, Alexander Kliche, Theo Verrips, Paul Kellam, Ariberto Fassati, Robin A Weiss
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-12-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1004552
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author Laura E McCoy
Lucy Rutten
Dan Frampton
Ian Anderson
Luke Granger
Rachael Bashford-Rogers
Gillian Dekkers
Nika M Strokappe
Michael S Seaman
Willie Koh
Vanina Grippo
Alexander Kliche
Theo Verrips
Paul Kellam
Ariberto Fassati
Robin A Weiss
author_facet Laura E McCoy
Lucy Rutten
Dan Frampton
Ian Anderson
Luke Granger
Rachael Bashford-Rogers
Gillian Dekkers
Nika M Strokappe
Michael S Seaman
Willie Koh
Vanina Grippo
Alexander Kliche
Theo Verrips
Paul Kellam
Ariberto Fassati
Robin A Weiss
author_sort Laura E McCoy
collection DOAJ
description To date, no immunization of humans or animals has elicited broadly neutralizing sera able to prevent HIV-1 transmission; however, elicitation of broad and potent heavy chain only antibodies (HCAb) has previously been reported in llamas. In this study, the anti-HIV immune responses in immunized llamas were studied via deep sequencing analysis using broadly neutralizing monoclonal HCAbs as a guides. Distinct neutralizing antibody lineages were identified in each animal, including two defined by novel antibodies (as variable regions called VHH) identified by robotic screening of over 6000 clones. The combined application of five VHH against viruses from clades A, B, C and CRF_AG resulted in neutralization as potent as any of the VHH individually and a predicted 100% coverage with a median IC50 of 0.17 µg/ml for the panel of 60 viruses tested. Molecular analysis of the VHH repertoires of two sets of immunized animals showed that each neutralizing lineage was only observed following immunization, demonstrating that they were elicited de novo. Our results show that immunization can induce potent and broadly neutralizing antibodies in llamas with features similar to human antibodies and provide a framework to analyze the effectiveness of immunization protocols.
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spelling doaj-art-83f2d3d6e15349dfb005fad45ed706272025-08-20T02:34:10ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742014-12-011012e100455210.1371/journal.ppat.1004552Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.Laura E McCoyLucy RuttenDan FramptonIan AndersonLuke GrangerRachael Bashford-RogersGillian DekkersNika M StrokappeMichael S SeamanWillie KohVanina GrippoAlexander KlicheTheo VerripsPaul KellamAriberto FassatiRobin A WeissTo date, no immunization of humans or animals has elicited broadly neutralizing sera able to prevent HIV-1 transmission; however, elicitation of broad and potent heavy chain only antibodies (HCAb) has previously been reported in llamas. In this study, the anti-HIV immune responses in immunized llamas were studied via deep sequencing analysis using broadly neutralizing monoclonal HCAbs as a guides. Distinct neutralizing antibody lineages were identified in each animal, including two defined by novel antibodies (as variable regions called VHH) identified by robotic screening of over 6000 clones. The combined application of five VHH against viruses from clades A, B, C and CRF_AG resulted in neutralization as potent as any of the VHH individually and a predicted 100% coverage with a median IC50 of 0.17 µg/ml for the panel of 60 viruses tested. Molecular analysis of the VHH repertoires of two sets of immunized animals showed that each neutralizing lineage was only observed following immunization, demonstrating that they were elicited de novo. Our results show that immunization can induce potent and broadly neutralizing antibodies in llamas with features similar to human antibodies and provide a framework to analyze the effectiveness of immunization protocols.https://doi.org/10.1371/journal.ppat.1004552
spellingShingle Laura E McCoy
Lucy Rutten
Dan Frampton
Ian Anderson
Luke Granger
Rachael Bashford-Rogers
Gillian Dekkers
Nika M Strokappe
Michael S Seaman
Willie Koh
Vanina Grippo
Alexander Kliche
Theo Verrips
Paul Kellam
Ariberto Fassati
Robin A Weiss
Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.
PLoS Pathogens
title Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.
title_full Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.
title_fullStr Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.
title_full_unstemmed Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.
title_short Molecular evolution of broadly neutralizing Llama antibodies to the CD4-binding site of HIV-1.
title_sort molecular evolution of broadly neutralizing llama antibodies to the cd4 binding site of hiv 1
url https://doi.org/10.1371/journal.ppat.1004552
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