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Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy

Interleukin-21 (IL-21), a member of the common cytokine receptor γ chain (γ(c)) family, is secreted by CD4(+) T cells and natural killer T cells and induces effector function through interactions with the IL-21 receptor (IL-21R)/γ(c) complex expressed on both immune and non-immune cells. Numerous st...

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Autores principales: Varkey, Reena, Du, Qun, Karnell, Jodi L., Xiao, Xiaodong, Casey, Kerry A., Woods, Rob, Rosenthal, Kim, Wilson, Susan, Dall’Acqua, William F., Wu, Herren, Herbst, Ronald, Ettinger, Rachel, Damschroder, Melissa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6347146/
https://www.ncbi.nlm.nih.gov/pubmed/30682117
http://dx.doi.org/10.1371/journal.pone.0211236
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author Varkey, Reena
Du, Qun
Karnell, Jodi L.
Xiao, Xiaodong
Casey, Kerry A.
Woods, Rob
Rosenthal, Kim
Wilson, Susan
Dall’Acqua, William F.
Wu, Herren
Herbst, Ronald
Ettinger, Rachel
Damschroder, Melissa
author_facet Varkey, Reena
Du, Qun
Karnell, Jodi L.
Xiao, Xiaodong
Casey, Kerry A.
Woods, Rob
Rosenthal, Kim
Wilson, Susan
Dall’Acqua, William F.
Wu, Herren
Herbst, Ronald
Ettinger, Rachel
Damschroder, Melissa
author_sort Varkey, Reena
collection PubMed
description Interleukin-21 (IL-21), a member of the common cytokine receptor γ chain (γ(c)) family, is secreted by CD4(+) T cells and natural killer T cells and induces effector function through interactions with the IL-21 receptor (IL-21R)/γ(c) complex expressed on both immune and non-immune cells. Numerous studies suggest that IL-21 plays a significant role in autoimmune disorders. Therapeutic intervention to disrupt the IL-21/IL-21R/γ(c) interaction and inhibit subsequent downstream signal transduction could offer a treatment paradigm for these diseases. Potent neutralizing antibodies reported in the literature were generated after extensive immunizations with human IL-21 alone and in combination with various adjuvants. To circumvent the laborious method of antibody generation while targeting a conserved functional epitope, we designed a novel alternating-antigen immunization strategy utilizing both human and cynomolgus monkey (cyno) IL-21. Despite the high degree of homology between human and cyno IL-21, our alternating-immunization strategy elicited higher antibody titers and more potent neutralizing hybridomas in mice than did the immunization with human IL-21 antigen alone. The lead hybridoma clone was humanized by grafting the murine complementarity-determining regions onto human germline framework templates, using a unique rational design. The final humanized and engineered antibody, MEDI7169, encodes only one murine residue at the variable heavy/light-chain interface, retains the sub-picomolar affinity for IL-21, specifically inhibits IL-21/IL-21R–mediated signaling events and is currently under clinical development as a potential therapeutic agent for autoimmune diseases. This study provides experimental evidence of the immune system’s potential to recognize and respond to shared epitopes of antigens from distinct species, and presents a generally applicable, novel method for the rapid generation of exceptional therapeutic antibodies using the hybridoma platform.
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spelling pubmed-63471462019-02-02 Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy Varkey, Reena Du, Qun Karnell, Jodi L. Xiao, Xiaodong Casey, Kerry A. Woods, Rob Rosenthal, Kim Wilson, Susan Dall’Acqua, William F. Wu, Herren Herbst, Ronald Ettinger, Rachel Damschroder, Melissa PLoS One Research Article Interleukin-21 (IL-21), a member of the common cytokine receptor γ chain (γ(c)) family, is secreted by CD4(+) T cells and natural killer T cells and induces effector function through interactions with the IL-21 receptor (IL-21R)/γ(c) complex expressed on both immune and non-immune cells. Numerous studies suggest that IL-21 plays a significant role in autoimmune disorders. Therapeutic intervention to disrupt the IL-21/IL-21R/γ(c) interaction and inhibit subsequent downstream signal transduction could offer a treatment paradigm for these diseases. Potent neutralizing antibodies reported in the literature were generated after extensive immunizations with human IL-21 alone and in combination with various adjuvants. To circumvent the laborious method of antibody generation while targeting a conserved functional epitope, we designed a novel alternating-antigen immunization strategy utilizing both human and cynomolgus monkey (cyno) IL-21. Despite the high degree of homology between human and cyno IL-21, our alternating-immunization strategy elicited higher antibody titers and more potent neutralizing hybridomas in mice than did the immunization with human IL-21 antigen alone. The lead hybridoma clone was humanized by grafting the murine complementarity-determining regions onto human germline framework templates, using a unique rational design. The final humanized and engineered antibody, MEDI7169, encodes only one murine residue at the variable heavy/light-chain interface, retains the sub-picomolar affinity for IL-21, specifically inhibits IL-21/IL-21R–mediated signaling events and is currently under clinical development as a potential therapeutic agent for autoimmune diseases. This study provides experimental evidence of the immune system’s potential to recognize and respond to shared epitopes of antigens from distinct species, and presents a generally applicable, novel method for the rapid generation of exceptional therapeutic antibodies using the hybridoma platform. Public Library of Science 2019-01-25 /pmc/articles/PMC6347146/ /pubmed/30682117 http://dx.doi.org/10.1371/journal.pone.0211236 Text en © 2019 Varkey et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Varkey, Reena
Du, Qun
Karnell, Jodi L.
Xiao, Xiaodong
Casey, Kerry A.
Woods, Rob
Rosenthal, Kim
Wilson, Susan
Dall’Acqua, William F.
Wu, Herren
Herbst, Ronald
Ettinger, Rachel
Damschroder, Melissa
Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy
title Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy
title_full Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy
title_fullStr Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy
title_full_unstemmed Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy
title_short Discovery and characterization of potent IL-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy
title_sort discovery and characterization of potent il-21 neutralizing antibodies via a novel alternating antigen immunization and humanization strategy
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6347146/
https://www.ncbi.nlm.nih.gov/pubmed/30682117
http://dx.doi.org/10.1371/journal.pone.0211236
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