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Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms

Autoantibodies against leucine-rich glioma inactivated 1 (LGI1) are found in patients with limbic encephalitis and focal seizures. Here, we generate patient-derived monoclonal antibodies (mAbs) against LGI1. We explore their sequences and binding characteristics, plus their pathogenic potential usin...

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Autores principales: Ramberger, Melanie, Berretta, Antonio, Tan, Jeanne M M, Sun, Bo, Michael, Sophia, Yeo, Tianrong, Theorell, Jakob, Bashford-Rogers, Rachael, Paneva, Sofija, O’Dowd, Victoria, Dedi, Neesha, Topia, Sarfaraj, Griffin, Robert, Ramirez-Franco, Jorge, El Far, Oussama, Baulac, Stéphanie, Leite, Maria I, Sen, Arjune, Jeans, Alexander, McMillan, David, Marshall, Diane, Anthony, Daniel, Lightwood, Daniel, Waters, Patrick, Irani, Sarosh R
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7296845/
https://www.ncbi.nlm.nih.gov/pubmed/32437528
http://dx.doi.org/10.1093/brain/awaa104
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author Ramberger, Melanie
Berretta, Antonio
Tan, Jeanne M M
Sun, Bo
Michael, Sophia
Yeo, Tianrong
Theorell, Jakob
Bashford-Rogers, Rachael
Paneva, Sofija
O’Dowd, Victoria
Dedi, Neesha
Topia, Sarfaraj
Griffin, Robert
Ramirez-Franco, Jorge
El Far, Oussama
Baulac, Stéphanie
Leite, Maria I
Sen, Arjune
Jeans, Alexander
McMillan, David
Marshall, Diane
Anthony, Daniel
Lightwood, Daniel
Waters, Patrick
Irani, Sarosh R
author_facet Ramberger, Melanie
Berretta, Antonio
Tan, Jeanne M M
Sun, Bo
Michael, Sophia
Yeo, Tianrong
Theorell, Jakob
Bashford-Rogers, Rachael
Paneva, Sofija
O’Dowd, Victoria
Dedi, Neesha
Topia, Sarfaraj
Griffin, Robert
Ramirez-Franco, Jorge
El Far, Oussama
Baulac, Stéphanie
Leite, Maria I
Sen, Arjune
Jeans, Alexander
McMillan, David
Marshall, Diane
Anthony, Daniel
Lightwood, Daniel
Waters, Patrick
Irani, Sarosh R
author_sort Ramberger, Melanie
collection PubMed
description Autoantibodies against leucine-rich glioma inactivated 1 (LGI1) are found in patients with limbic encephalitis and focal seizures. Here, we generate patient-derived monoclonal antibodies (mAbs) against LGI1. We explore their sequences and binding characteristics, plus their pathogenic potential using transfected HEK293T cells, rodent neuronal preparations, and behavioural and electrophysiological assessments in vivo after mAb injections into the rodent hippocampus. In live cell-based assays, LGI1 epitope recognition was examined with patient sera (n = 31), CSFs (n = 11), longitudinal serum samples (n = 15), and using mAbs (n = 14) generated from peripheral B cells of two patients. All sera and 9/11 CSFs bound both the leucine-rich repeat (LRR) and the epitempin repeat (EPTP) domains of LGI1, with stable ratios of LRR:EPTP antibody levels over time. By contrast, the mAbs derived from both patients recognized either the LRR or EPTP domain. mAbs against both domain specificities showed varied binding strengths, and marked genetic heterogeneity, with high mutation frequencies. LRR-specific mAbs recognized LGI1 docked to its interaction partners, ADAM22 and ADAM23, bound to rodent brain sections, and induced internalization of the LGI1-ADAM22/23 complex in both HEK293T cells and live hippocampal neurons. By contrast, few EPTP-specific mAbs bound to rodent brain sections or ADAM22/23-docked LGI1, but all inhibited the docking of LGI1 to ADAM22/23. After intrahippocampal injection, and by contrast to the LRR-directed mAbs, the EPTP-directed mAbs showed far less avid binding to brain tissue and were consistently detected in the serum. Post-injection, both domain-specific mAbs abrogated long-term potentiation induction, and LRR-directed antibodies with higher binding strengths induced memory impairment. Taken together, two largely dichotomous populations of LGI1 mAbs with distinct domain binding characteristics exist in the affinity matured peripheral autoantigen-specific memory pools of individuals, both of which have pathogenic potential. In human autoantibody-mediated diseases, the detailed characterization of patient mAbs provides a valuable method to dissect the molecular mechanisms within polyclonal populations.
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spelling pubmed-72968452020-06-22 Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms Ramberger, Melanie Berretta, Antonio Tan, Jeanne M M Sun, Bo Michael, Sophia Yeo, Tianrong Theorell, Jakob Bashford-Rogers, Rachael Paneva, Sofija O’Dowd, Victoria Dedi, Neesha Topia, Sarfaraj Griffin, Robert Ramirez-Franco, Jorge El Far, Oussama Baulac, Stéphanie Leite, Maria I Sen, Arjune Jeans, Alexander McMillan, David Marshall, Diane Anthony, Daniel Lightwood, Daniel Waters, Patrick Irani, Sarosh R Brain Original Articles Autoantibodies against leucine-rich glioma inactivated 1 (LGI1) are found in patients with limbic encephalitis and focal seizures. Here, we generate patient-derived monoclonal antibodies (mAbs) against LGI1. We explore their sequences and binding characteristics, plus their pathogenic potential using transfected HEK293T cells, rodent neuronal preparations, and behavioural and electrophysiological assessments in vivo after mAb injections into the rodent hippocampus. In live cell-based assays, LGI1 epitope recognition was examined with patient sera (n = 31), CSFs (n = 11), longitudinal serum samples (n = 15), and using mAbs (n = 14) generated from peripheral B cells of two patients. All sera and 9/11 CSFs bound both the leucine-rich repeat (LRR) and the epitempin repeat (EPTP) domains of LGI1, with stable ratios of LRR:EPTP antibody levels over time. By contrast, the mAbs derived from both patients recognized either the LRR or EPTP domain. mAbs against both domain specificities showed varied binding strengths, and marked genetic heterogeneity, with high mutation frequencies. LRR-specific mAbs recognized LGI1 docked to its interaction partners, ADAM22 and ADAM23, bound to rodent brain sections, and induced internalization of the LGI1-ADAM22/23 complex in both HEK293T cells and live hippocampal neurons. By contrast, few EPTP-specific mAbs bound to rodent brain sections or ADAM22/23-docked LGI1, but all inhibited the docking of LGI1 to ADAM22/23. After intrahippocampal injection, and by contrast to the LRR-directed mAbs, the EPTP-directed mAbs showed far less avid binding to brain tissue and were consistently detected in the serum. Post-injection, both domain-specific mAbs abrogated long-term potentiation induction, and LRR-directed antibodies with higher binding strengths induced memory impairment. Taken together, two largely dichotomous populations of LGI1 mAbs with distinct domain binding characteristics exist in the affinity matured peripheral autoantigen-specific memory pools of individuals, both of which have pathogenic potential. In human autoantibody-mediated diseases, the detailed characterization of patient mAbs provides a valuable method to dissect the molecular mechanisms within polyclonal populations. Oxford University Press 2020-06 2020-05-21 /pmc/articles/PMC7296845/ /pubmed/32437528 http://dx.doi.org/10.1093/brain/awaa104 Text en © The Author(s) (2020). Published by Oxford University Press on behalf of the Guarantors of Brain. 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 reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Ramberger, Melanie
Berretta, Antonio
Tan, Jeanne M M
Sun, Bo
Michael, Sophia
Yeo, Tianrong
Theorell, Jakob
Bashford-Rogers, Rachael
Paneva, Sofija
O’Dowd, Victoria
Dedi, Neesha
Topia, Sarfaraj
Griffin, Robert
Ramirez-Franco, Jorge
El Far, Oussama
Baulac, Stéphanie
Leite, Maria I
Sen, Arjune
Jeans, Alexander
McMillan, David
Marshall, Diane
Anthony, Daniel
Lightwood, Daniel
Waters, Patrick
Irani, Sarosh R
Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms
title Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms
title_full Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms
title_fullStr Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms
title_full_unstemmed Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms
title_short Distinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanisms
title_sort distinctive binding properties of human monoclonal lgi1 autoantibodies determine pathogenic mechanisms
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7296845/
https://www.ncbi.nlm.nih.gov/pubmed/32437528
http://dx.doi.org/10.1093/brain/awaa104
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