Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration.
OBJECTIVE
To determine sensitivity and specificity of a standardized recombinant cell-based indirect immunofluorescence assay (RC-IFA) for anti-Tr antibodies in comparison to a reference procedure.


METHODS
Delta/Notch-like epidermal growth factor-related receptor (DNER) was expressed in HEK293 and used as a substrate for RC-IFA. HEK293 control cells expressing CDR2/Yo and CDR2L as well as mock-transfected HEK293 cells were used as controls. Serum samples from 38 patients with anti-Tr antibodies (33 with paraneoplastic cerebellar degeneration [PCD] and Hodgkin lymphoma), 66 patients with anti-Tr-negative PCD, 53 patients with Hodgkin lymphoma without neurologic symptoms, 40 patients with rheumatic diseases, and 42 healthy blood donors were tested for anti-DNER reactivity in the RC-IFA. In addition, RC-IFA results were compared to those from a commercial tissue-based IFA using monkey cerebellum.


RESULTS
Using the RC-IFA, anti-DNER was detected in all anti-Tr-positive patients but in none of the controls (sensitivity 100%, 95% confidence interval [CI] 92.8%-100%; specificity 100%, 95% CI 98.7%-100%). In comparison, anti-Tr was not detected in 4 samples with low-titer autoantibodies using the commercial tissue-based assay. Preadsorption of sera with either recombinant full-length DNER or its extracellular domain selectively abolished anti-Tr reactivity.


CONCLUSIONS
Anti-Tr antibodies bind to the extracellular domain of DNER and can be detected by RC-IFA using HEK293 cells expressing the recombinant receptor. The new method performs better than a frequently used commercial tissue-based indirect immunofluorescence assay (IFA) in samples with low-titer antibodies.


METHODS
This study provides Class II evidence that RC-IFA accurately detects anti-Tr as compared to conventional IFA.
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Christian
C
Probst
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Lars
L
Komorowski
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Esther
E
de Graaff
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Marleen
M
van Coevorden-Hameete
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Veronique
V
Rogemond
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Jerome
J
Honnorat
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Lidia
L
Sabeter
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Francesc
F
Graus
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Sven
S
Jarius
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Raymond
R
Voltz
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Brigitte
B
Wildemann
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Diego
D
Franciotta
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Inga M
IM
Blöcker
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Wolfgang
W
Schlumberger
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Winfried
W
Stöcker
Institute of Experimental Immunology (C.P., L.K., I.M.B., W. Schlumberger, W. Stöcker), Euroimmun AG, Lübeck, Germany; Department of Biology (E.d.G., M.v.C.-H.), Division of Cell Biology, Utrecht University, the Netherlands; Department of Neurology (M.v.C.-H., P.A.E.S.S.), Erasmus MC, Rotterdam, the Netherlands; Neuro-oncology Department (V.R., J.H.), French Paraneoplastic Neurological Syndrome Reference Center, Hospices Civils de Lyon, Hôpital Neurologique, Bron, France; Lyon Neuroscience Research Center (V.R., J.H.), INSERM U1028/CNRS UMR 5292, Lyon, France; University of Claude Bernard (V.R., J.H.), Lyon 1, Villeurbanne, France; Institut d' Investigació Biomèdica August Pi i Sunyer (IDIBAPS) (L.S., F.G.), Barcelona, Spain; Department of Molecular Neuroimmunology (S.J., B.W.), University of Heidelberg, Germany; Department of Palliative Medicine (R.V.), University Hospital Cologne, Germany; and IRCCS (D.F.), C. Mondino National Neurological Institute, Pavia, Italy.
Peter A E
PA
Sillevis Smitt