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Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro

Subvisible proteinaceous particles which are present in all therapeutic protein formulations are in the focus of intense discussions between health authorities, academics and biopharmaceutical companies in the context of concerns that such particles could promote unwanted immunogenicity via anti-dru...

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Autores principales: Rombach-Riegraf, Verena, Karle, Anette C., Wolf, Babette, Sordé, Laetitia, Koepke, Stephan, Gottlieb, Sascha, Krieg, Jennifer, Djidja, Marie-Claude, Baban, Aida, Spindeldreher, Sebastian, Koulov, Atanas V., Kiessling, Andrea
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3897673/
https://www.ncbi.nlm.nih.gov/pubmed/24466023
http://dx.doi.org/10.1371/journal.pone.0086322
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author Rombach-Riegraf, Verena
Karle, Anette C.
Wolf, Babette
Sordé, Laetitia
Koepke, Stephan
Gottlieb, Sascha
Krieg, Jennifer
Djidja, Marie-Claude
Baban, Aida
Spindeldreher, Sebastian
Koulov, Atanas V.
Kiessling, Andrea
author_facet Rombach-Riegraf, Verena
Karle, Anette C.
Wolf, Babette
Sordé, Laetitia
Koepke, Stephan
Gottlieb, Sascha
Krieg, Jennifer
Djidja, Marie-Claude
Baban, Aida
Spindeldreher, Sebastian
Koulov, Atanas V.
Kiessling, Andrea
author_sort Rombach-Riegraf, Verena
collection PubMed
description Subvisible proteinaceous particles which are present in all therapeutic protein formulations are in the focus of intense discussions between health authorities, academics and biopharmaceutical companies in the context of concerns that such particles could promote unwanted immunogenicity via anti-drug antibody formation. In order to provide further understanding of the subject, this study closely examines the specific biological effects proteinaceous particles may exert on dendritic cells (DCs) as the most efficient antigen-presenting cell population crucial for the initiation of the adaptive immune response. Two different model IgG antibodies were subjected to three different types of exaggerated physical stress to generate subvisible particles in far greater concentrations than the ones typical for the currently marketed biotherapeutical antibodies. The aggregated samples were used in in vitro biological assays in order to interrogate the early DC-driven events that initiate CD4 T-cell dependent humoral adaptive immune responses – peptide presentation capacity and co-stimulatory activity of DCs. Most importantly, antigen presentation was addressed with a unique approach called MHC-associated Peptide Proteomics (MAPPs), which allows for identifying the sequences of HLA-DR associated peptides directly from human dendritic cells. The experiments demonstrated that highly aggregated solutions of two model mAbs generated under controlled conditions can induce activation of human monocyte-derived DCs as indicated by upregulation of typical maturation markers including co-stimulatory molecules necessary for CD4 T-cell activation. Additional data suggest that highly aggregated proteins could induce in vitro T-cell responses. Intriguingly, strong aggregation-mediated changes in the pattern and quantity of antigen-derived HLA-DR associated peptides presented on DCs were observed, indicating a change in protein processing and presentation. Increasing the amounts of subvisible proteinaceous particles correlated very well with the pronounced increase in the peptide number and clusters presented in the context of class II HLA-DR molecules, suggesting a major involvement of a mass-action mechanism of altering the presentation.
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spelling pubmed-38976732014-01-24 Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro Rombach-Riegraf, Verena Karle, Anette C. Wolf, Babette Sordé, Laetitia Koepke, Stephan Gottlieb, Sascha Krieg, Jennifer Djidja, Marie-Claude Baban, Aida Spindeldreher, Sebastian Koulov, Atanas V. Kiessling, Andrea PLoS One Research Article Subvisible proteinaceous particles which are present in all therapeutic protein formulations are in the focus of intense discussions between health authorities, academics and biopharmaceutical companies in the context of concerns that such particles could promote unwanted immunogenicity via anti-drug antibody formation. In order to provide further understanding of the subject, this study closely examines the specific biological effects proteinaceous particles may exert on dendritic cells (DCs) as the most efficient antigen-presenting cell population crucial for the initiation of the adaptive immune response. Two different model IgG antibodies were subjected to three different types of exaggerated physical stress to generate subvisible particles in far greater concentrations than the ones typical for the currently marketed biotherapeutical antibodies. The aggregated samples were used in in vitro biological assays in order to interrogate the early DC-driven events that initiate CD4 T-cell dependent humoral adaptive immune responses – peptide presentation capacity and co-stimulatory activity of DCs. Most importantly, antigen presentation was addressed with a unique approach called MHC-associated Peptide Proteomics (MAPPs), which allows for identifying the sequences of HLA-DR associated peptides directly from human dendritic cells. The experiments demonstrated that highly aggregated solutions of two model mAbs generated under controlled conditions can induce activation of human monocyte-derived DCs as indicated by upregulation of typical maturation markers including co-stimulatory molecules necessary for CD4 T-cell activation. Additional data suggest that highly aggregated proteins could induce in vitro T-cell responses. Intriguingly, strong aggregation-mediated changes in the pattern and quantity of antigen-derived HLA-DR associated peptides presented on DCs were observed, indicating a change in protein processing and presentation. Increasing the amounts of subvisible proteinaceous particles correlated very well with the pronounced increase in the peptide number and clusters presented in the context of class II HLA-DR molecules, suggesting a major involvement of a mass-action mechanism of altering the presentation. Public Library of Science 2014-01-21 /pmc/articles/PMC3897673/ /pubmed/24466023 http://dx.doi.org/10.1371/journal.pone.0086322 Text en © 2014 Rombach-Riegraf 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, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Rombach-Riegraf, Verena
Karle, Anette C.
Wolf, Babette
Sordé, Laetitia
Koepke, Stephan
Gottlieb, Sascha
Krieg, Jennifer
Djidja, Marie-Claude
Baban, Aida
Spindeldreher, Sebastian
Koulov, Atanas V.
Kiessling, Andrea
Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro
title Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro
title_full Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro
title_fullStr Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro
title_full_unstemmed Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro
title_short Aggregation of Human Recombinant Monoclonal Antibodies Influences the Capacity of Dendritic Cells to Stimulate Adaptive T-Cell Responses In Vitro
title_sort aggregation of human recombinant monoclonal antibodies influences the capacity of dendritic cells to stimulate adaptive t-cell responses in vitro
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3897673/
https://www.ncbi.nlm.nih.gov/pubmed/24466023
http://dx.doi.org/10.1371/journal.pone.0086322
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