Identification of delta/notch‐like epidermal growth factor‐related receptor as the Tr antigen in paraneoplastic cerebellar degeneration

• DOI: 10.1002/ana.23550
• Publication Date: 2012-02-01T15:29:41Z
• Authors: Esther de Graaff; Peter Maat; Esther Hulsenboom; Robert van den Berg; Martin van den Bent; Jeroen Demmers; Pieternella J. Lu...; Casper C. Hoogenraad; Peter Sillevis Smitt
• Keywords: Paraneoplastic cerebellar degeneration

Abstract Objective: Anti‐Tr is among the better described autoantibodies in paraneoplastic cerebellar degeneration (PCD) combined with Hodgkin lymphoma (HL); however, Tr antigen remains unidentified. Methods: We used immunoprecipitation of total rat brain extract followed by mass spectrometry to identify recognized anti‐Tr–positive sera. By Western blotting and cell‐based assays, we tested a 12 246 control sera determined region epitope anti‐Tr antibodies. Deletion mutant constructs were generated further map antigenic region. Results: Mass analysis immunopurified using 4 different led identification Delta/Notch‐like epidermal growth factor‐related receptor (DNER) as antigen. All but 1 samples negative HeLa screening assay, whereas stained hemagglutinin‐tagged DNER‐expressing cells. Only subject HL no ataxia was found be both DNER positive. Using deletion constructs, pinpointed main extracellular domain. Knockdown endogenous hippocampal N‐glycosylation mutations abolished staining, indicating that glycosylation required for it Interpretation: detected Presence antibodies patients PCD or only can now screened quickly reliably assay. ANN NEUROL 2012