Peripherally derived LGI1-reactive monoclonal antibodies cause epileptic seizuresin vivo

• DOI: 10.1101/2023.10.11.561725
• Publication Date: 2023-10-16T17:20:26Z
• Authors: Manoj Upadhya; Toni Kirmann; Max Wilson; Christian M Simon; Divya Dhangar; Christian Geis; Robyn Williams; Gavin Woodhall; Stefan Hallermann; Sarosh R Irani; Sukhvir K Wright
• Keywords: Male; Epilepsy; Seizures; Report; Intracellular Signaling Peptides and Proteins; Animals; Antibodies, Monoclonal; Humans; Proteins; Electroencephalography; Rats, Wistar; Rats; Autoantibodies

AbstractOne striking clinical hallmark in patients with autoantibodies to leucine-rich glioma inactivated 1 (LGI1) is the very frequent focal seizure semiologies, including faciobrachial dystonic seizures (FBDS), in addition to the amnesia. Polyclonal serum IgGs have successfully modelled the cognitive changesin vivobut not seizures. Hence, it remains unclear whether LGI1-autoantibodies are sufficient to cause seizures.We tested this with the molecularly precise monoclonal antibodies directed against LGI1 (LGI1-mAbs), derived from patient circulating B cells. These were directed towards both major domains of LGI1, LRR (n=5) and EPTP (n=5) and infused intracerebroventricularly over 7 days into juvenile male Wistar rats using osmotic pumps. Continuous wireless EEG was recorded from a depth electrode placed in hippocampal CA3 plus behavioural tests for memory and hyperexcitability were performed. Following infusion completion (Day 9), post-mortem brain slices were studied using electrophysiology and immunostaining.By comparison to control-mAb injected rats (n=6), video-EEG analysis over 9 days revealed convulsive and non-convulsive seizure activity in rats infused with LGI1-mAbs, with a significant number of ictal events (245±83 vs. 7.8±7.8 in controls; p=0.002). Memory was not impaired in the novel object recognition test. Local field potential recordings from postmortem brain slices showed spontaneous ictal-like spike activity in the CA3 region (p=0.03). The LGI1-mAbs bound most strongly in the hippocampal CA3 region and induced a significant reduction in Kv1.1 cluster number in this subfield (6 controls; 7 LGI1-mAbs; p=0.01)Peripherally-derived human LGI1-mAbs infused into rodent CSF provide strong evidence of directin vivoepileptogenesis with molecular correlations. These findings fulfill criteria for LGI1-antibodies in seizure causation.