Identification of DAGLA as an autoantibody target in cerebellar ataxia

Author:

Miske RamonaORCID,Scharf Madeleine,Borowski Kathrin,Specht Ina,Corty Merle,Loritz Monika-Johanna,Rombach Frederik,Laureys Guy,Rochow Nadine,Radzimski Christiane,Schnitter Linda,Ratuszny Dominica,Skripuletz ThomasORCID,Wattjes Mike PORCID,Hahn Stefanie,Denno Yvonne,Guerti Khadija,Oyaert Matthijs,Benkhadra Farid,Bien Corinna Ines,Nitsch Sophie,Wandinger Klaus-Peter,van Pesch Vincent,Probst Christian,Teegen Bianca,Komorowski Lars,Sühs Kurt-Wolfram

Abstract

BackgroundWe aimed to investigate the clinical, imaging and fluid biomarker characteristics in patients with antidiacylglycerol lipase alpha (DAGLA)-autoantibody-associated cerebellitis.MethodsSerum and cerebrospinal fliud (CSF) samples from four index patients were subjected to comprehensive autoantibody screening by indirect immunofluorescence assay (IIFA). Immunoprecipitation, mass spectrometry and recombinant protein assays were used to identify the autoantigen. Sera from 101 patients with various neurological symptoms and a similar tissue staining pattern as the index patient samples, and 102 healthy donors were analysed in recombinant cell-based IIFA (RC-IIFA) with the identified protein. Epitope characterisation of all positive samples was performed via ELISA, immunoblot, immunoprecipitation and RC-IIFA using different DAGLA fragments.ResultsAll index patients were relatively young (age: 18–34) and suffered from pronounced gait ataxia, dysarthria and visual impairments. Paraclinical hallmarks in early-stage disease were inflammatory CSF changes and cerebellar cortex hyperintensity in MRI. Severe cerebellar atrophy developed in three of four patients within 6 months. All patient samples showed the same unclassified IgG reactivity with the cerebellar molecular layer. DAGLA was identified as the target antigen and confirmed by competitive inhibition experiments and DAGLA-specific RC-IIFA. In RC-IIFA, serum reactivity against DAGLA was also found in 17/101 disease controls, including patients with different clinical phenotypes than the one of the index patients, and in 1/102 healthy donors. Epitope characterisation revealed that 17/18 anti-DAGLA-positive control sera reacted with a C-terminal intracellular DAGLA 583–1042 fragment, while the CSF samples of the index patients targeted a conformational epitope between amino acid 1 and 157.ConclusionsWe propose that anti-DAGLA autoantibodies detected in CSF, with a characteristic tissue IIFA pattern, represent novel biomarkers for rapidly progressive cerebellitis.

Publisher

BMJ

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