Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration

Christian Probst; Lars Komorowski; Esther de Graaff; Marleen van Coevorden-Hameete; Veronique Rogemond; Jerome Honnorat; Lidia Sabeter; Francesc Graus; Sven Jarius; Raymond Voltz; Brigitte Wildemann; Diego Franciotta; Inga M Blöcker; Wolfgang Schlumberger; Winfried Stöcker; Peter A E Sillevis Smitt

doi:10.1212/NXI.0000000000000068

Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration

Christian Probst
, Lars Komorowski
, Esther de Graaff
, Marleen van Coevorden-Hameete
, Veronique Rogemond
, Jerome Honnorat
, Lidia Sabeter
, Francesc Graus
, Sven Jarius
, Raymond Voltz
, Brigitte Wildemann
, Diego Franciotta
, Inga M Blöcker
, Wolfgang Schlumberger
, Winfried Stöcker
, Peter A E Sillevis Smitt

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

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.

CONCLUSION: 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.

CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that RC-IFA accurately detects anti-Tr as compared to conventional IFA.

Original language	English
Article number	e68
Number of pages	7
Journal	Neurology® neuroimmunology & neuroinflammation
Volume	2
Issue number	2
DOIs	https://doi.org/10.1212/NXI.0000000000000068
Publication status	Published - Apr 2015

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Probst, C., Komorowski, L., de Graaff, E., van Coevorden-Hameete, M., Rogemond, V., Honnorat, J., Sabeter, L., Graus, F., Jarius, S., Voltz, R., Wildemann, B., Franciotta, D., Blöcker, I. M., Schlumberger, W., Stöcker, W., & Sillevis Smitt, P. A. E. (2015). Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration. Neurology® neuroimmunology & neuroinflammation, 2(2), Article e68. https://doi.org/10.1212/NXI.0000000000000068

@article{425294b28a5b471db5881c562836faab,

title = "Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration",

abstract = "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.CONCLUSION: 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.CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that RC-IFA accurately detects anti-Tr as compared to conventional IFA.",

author = "Christian Probst and Lars Komorowski and \{de Graaff\}, Esther and \{van Coevorden-Hameete\}, Marleen and Veronique Rogemond and Jerome Honnorat and Lidia Sabeter and Francesc Graus and Sven Jarius and Raymond Voltz and Brigitte Wildemann and Diego Franciotta and Bl{\"o}cker, \{Inga M\} and Wolfgang Schlumberger and Winfried St{\"o}cker and \{Sillevis Smitt\}, \{Peter A E\}",

year = "2015",

month = apr,

doi = "10.1212/NXI.0000000000000068",

language = "English",

volume = "2",

journal = "Neurology{\textregistered} neuroimmunology \& neuroinflammation",

issn = "2332-7812",

publisher = "Lippincott Williams and Wilkins",

number = "2",

}

Probst, C, Komorowski, L, de Graaff, E, van Coevorden-Hameete, M, Rogemond, V, Honnorat, J, Sabeter, L, Graus, F, Jarius, S, Voltz, R, Wildemann, B, Franciotta, D, Blöcker, IM, Schlumberger, W, Stöcker, W & Sillevis Smitt, PAE 2015, 'Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration', Neurology® neuroimmunology & neuroinflammation, vol. 2, no. 2, e68. https://doi.org/10.1212/NXI.0000000000000068

TY - JOUR

T1 - Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration

AU - Probst, Christian

AU - Komorowski, Lars

AU - de Graaff, Esther

AU - van Coevorden-Hameete, Marleen

AU - Rogemond, Veronique

AU - Honnorat, Jerome

AU - Sabeter, Lidia

AU - Graus, Francesc

AU - Jarius, Sven

AU - Voltz, Raymond

AU - Wildemann, Brigitte

AU - Franciotta, Diego

AU - Blöcker, Inga M

AU - Schlumberger, Wolfgang

AU - Stöcker, Winfried

AU - Sillevis Smitt, Peter A E

PY - 2015/4

Y1 - 2015/4

N2 - 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.CONCLUSION: 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.CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that RC-IFA accurately detects anti-Tr as compared to conventional IFA.

AB - 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.CONCLUSION: 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.CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that RC-IFA accurately detects anti-Tr as compared to conventional IFA.

U2 - 10.1212/NXI.0000000000000068

DO - 10.1212/NXI.0000000000000068

M3 - Article

C2 - 25745634

SN - 2332-7812

VL - 2

JO - Neurology® neuroimmunology & neuroinflammation

JF - Neurology® neuroimmunology & neuroinflammation

IS - 2

M1 - e68

ER -

Standardized test for anti-Tr/DNER in patients with paraneoplastic cerebellar degeneration

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