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...
| Hoofdauteurs: | , , , , , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Formaat: | Journal article |
| Taal: | English |
| Gepubliceerd in: |
Oxford University Press
2020
|
| _version_ | 1826260986291552256 |
|---|---|
| author | Ramberger, M Berretta, A Tan, JMM Sun, B Michael, S Yeo, T Theorell, J Bashford-Rogers, R Paneva, S O'Dowd, V Dedi, N Topia, S Griffin, R Ramirez-Franco, J El Far, O Baulac, S Leite, MI Sen, A Jeans, A McMillan, D Marshall, D Anthony, D Lightwood, D Waters, P Irani, SR |
| author_facet | Ramberger, M Berretta, A Tan, JMM Sun, B Michael, S Yeo, T Theorell, J Bashford-Rogers, R Paneva, S O'Dowd, V Dedi, N Topia, S Griffin, R Ramirez-Franco, J El Far, O Baulac, S Leite, MI Sen, A Jeans, A McMillan, D Marshall, D Anthony, D Lightwood, D Waters, P Irani, SR |
| author_sort | Ramberger, M |
| collection | OXFORD |
| 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. |
| first_indexed | 2024-03-06T19:14:26Z |
| format | Journal article |
| id | oxford-uuid:17de9f6d-7a20-41f9-aeff-1e17c7d63ea9 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:14:26Z |
| publishDate | 2020 |
| publisher | Oxford University Press |
| record_format | dspace |
| spelling | oxford-uuid:17de9f6d-7a20-41f9-aeff-1e17c7d63ea92022-03-26T10:39:59ZDistinctive binding properties of human monoclonal LGI1 autoantibodies determine pathogenic mechanismsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:17de9f6d-7a20-41f9-aeff-1e17c7d63ea9EnglishSymplectic ElementsOxford University Press2020Ramberger, MBerretta, ATan, JMMSun, BMichael, SYeo, TTheorell, JBashford-Rogers, RPaneva, SO'Dowd, VDedi, NTopia, SGriffin, RRamirez-Franco, JEl Far, OBaulac, SLeite, MISen, AJeans, AMcMillan, DMarshall, DAnthony, DLightwood, DWaters, PIrani, SRAutoantibodies 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. |
| spellingShingle | Ramberger, M Berretta, A Tan, JMM Sun, B Michael, S Yeo, T Theorell, J Bashford-Rogers, R Paneva, S O'Dowd, V Dedi, N Topia, S Griffin, R Ramirez-Franco, J El Far, O Baulac, S Leite, MI Sen, A Jeans, A McMillan, D Marshall, D Anthony, D Lightwood, D Waters, P Irani, SR 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 |
| work_keys_str_mv | AT rambergerm distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT berrettaa distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT tanjmm distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT sunb distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT michaels distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT yeot distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT theorellj distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT bashfordrogersr distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT panevas distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT odowdv distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT dedin distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT topias distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT griffinr distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT ramirezfrancoj distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT elfaro distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT baulacs distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT leitemi distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT sena distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT jeansa distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT mcmilland distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT marshalld distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT anthonyd distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT lightwoodd distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT watersp distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms AT iranisr distinctivebindingpropertiesofhumanmonoclonallgi1autoantibodiesdeterminepathogenicmechanisms |