Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus

Christian Probst; Wolfgang Schlumberger; Winfried Stöcker; Andreas Recke; Enno Schmidt; Takashi Hashimoto; Xue Jun Zhu; Detlef Zillikens; Lars Komorowski

doi:10.1016/j.cca.2009.08.022

Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus

Christian Probst, Wolfgang Schlumberger, Winfried Stöcker, Andreas Recke, Enno Schmidt, Takashi Hashimoto, Xue Jun Zhu, Detlef Zillikens, Lars Komorowski

Abstract

INTRODUCTION: In paraneoplastic pemphigus (PNP), indirect immunofluorescence microscopy using rat bladder sections is widely used to search for circulating autoantibodies which are predominantly directed against envoplakin and periplakin. A sensitive and specific detection system for envoplakin/periplakin-specific autoantibodies is not yet available.

METHODS: Overlapping fragments spanning envoplakin and periplakin, respectively, were analyzed for their ability to bind PNP-specific autoantibodies by immunoblotting and ELISA in sera from patients with PNP (n=31), pemphigus vulgaris (n=30), and bullous pemphigoid (n=50) as well as healthy volunteers (n=140). The results were compared with those obtained by immunoblotting of extract of cultured human keratinocytes.

RESULTS: Immunoblot analysis revealed that most sera contained antibodies against the N-termini of both plakins as well as against the C-terminus of envoplakin. By ELISA, reactivities against envoplakin(1-481), envoplakin(1626-2033), and periplakin(1-324) were found in 25 (80.6%), 25 (80.6%), and 23 (74.2%) PNP sera, and in 1 (1.2%), 3 (3.7%), and 2 (2.5%) control sera, respectively.

CONCLUSIONS: The new ELISA based on an N-terminal fragment of envoplakin shows a high diagnostic accuracy to detect circulating autoantibodies in PNP. It is easy to be setup and standardized and can help to differentiate PNP patients from those with pemphigus vulgaris.

Original language	English
Journal	Clinica Chimica Acta
Volume	410
Issue number	1-2
Pages (from-to)	13-8
Number of pages	6
ISSN	0009-8981
DOIs	https://doi.org/10.1016/j.cca.2009.08.022
Publication status	Published - 2009

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10.1016/j.cca.2009.08.022

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@article{3df4c25bdb6a4486bdb1b2b732cb9e62,

title = "Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus",

abstract = "INTRODUCTION: In paraneoplastic pemphigus (PNP), indirect immunofluorescence microscopy using rat bladder sections is widely used to search for circulating autoantibodies which are predominantly directed against envoplakin and periplakin. A sensitive and specific detection system for envoplakin/periplakin-specific autoantibodies is not yet available.METHODS: Overlapping fragments spanning envoplakin and periplakin, respectively, were analyzed for their ability to bind PNP-specific autoantibodies by immunoblotting and ELISA in sera from patients with PNP (n=31), pemphigus vulgaris (n=30), and bullous pemphigoid (n=50) as well as healthy volunteers (n=140). The results were compared with those obtained by immunoblotting of extract of cultured human keratinocytes.RESULTS: Immunoblot analysis revealed that most sera contained antibodies against the N-termini of both plakins as well as against the C-terminus of envoplakin. By ELISA, reactivities against envoplakin(1-481), envoplakin(1626-2033), and periplakin(1-324) were found in 25 (80.6%), 25 (80.6%), and 23 (74.2%) PNP sera, and in 1 (1.2%), 3 (3.7%), and 2 (2.5%) control sera, respectively.CONCLUSIONS: The new ELISA based on an N-terminal fragment of envoplakin shows a high diagnostic accuracy to detect circulating autoantibodies in PNP. It is easy to be setup and standardized and can help to differentiate PNP patients from those with pemphigus vulgaris.",

author = "Christian Probst and Wolfgang Schlumberger and Winfried St{\"o}cker and Andreas Recke and Enno Schmidt and Takashi Hashimoto and Zhu, {Xue Jun} and Detlef Zillikens and Lars Komorowski",

year = "2009",

doi = "10.1016/j.cca.2009.08.022",

language = "English",

volume = "410",

pages = "13--8",

journal = "Clinica Chimica Acta",

issn = "0009-8981",

number = "1-2",

}

TY - JOUR

T1 - Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus

AU - Probst, Christian

AU - Schlumberger, Wolfgang

AU - Stöcker, Winfried

AU - Recke, Andreas

AU - Schmidt, Enno

AU - Hashimoto, Takashi

AU - Zhu, Xue Jun

AU - Zillikens, Detlef

AU - Komorowski, Lars

PY - 2009

Y1 - 2009

N2 - INTRODUCTION: In paraneoplastic pemphigus (PNP), indirect immunofluorescence microscopy using rat bladder sections is widely used to search for circulating autoantibodies which are predominantly directed against envoplakin and periplakin. A sensitive and specific detection system for envoplakin/periplakin-specific autoantibodies is not yet available.METHODS: Overlapping fragments spanning envoplakin and periplakin, respectively, were analyzed for their ability to bind PNP-specific autoantibodies by immunoblotting and ELISA in sera from patients with PNP (n=31), pemphigus vulgaris (n=30), and bullous pemphigoid (n=50) as well as healthy volunteers (n=140). The results were compared with those obtained by immunoblotting of extract of cultured human keratinocytes.RESULTS: Immunoblot analysis revealed that most sera contained antibodies against the N-termini of both plakins as well as against the C-terminus of envoplakin. By ELISA, reactivities against envoplakin(1-481), envoplakin(1626-2033), and periplakin(1-324) were found in 25 (80.6%), 25 (80.6%), and 23 (74.2%) PNP sera, and in 1 (1.2%), 3 (3.7%), and 2 (2.5%) control sera, respectively.CONCLUSIONS: The new ELISA based on an N-terminal fragment of envoplakin shows a high diagnostic accuracy to detect circulating autoantibodies in PNP. It is easy to be setup and standardized and can help to differentiate PNP patients from those with pemphigus vulgaris.

AB - INTRODUCTION: In paraneoplastic pemphigus (PNP), indirect immunofluorescence microscopy using rat bladder sections is widely used to search for circulating autoantibodies which are predominantly directed against envoplakin and periplakin. A sensitive and specific detection system for envoplakin/periplakin-specific autoantibodies is not yet available.METHODS: Overlapping fragments spanning envoplakin and periplakin, respectively, were analyzed for their ability to bind PNP-specific autoantibodies by immunoblotting and ELISA in sera from patients with PNP (n=31), pemphigus vulgaris (n=30), and bullous pemphigoid (n=50) as well as healthy volunteers (n=140). The results were compared with those obtained by immunoblotting of extract of cultured human keratinocytes.RESULTS: Immunoblot analysis revealed that most sera contained antibodies against the N-termini of both plakins as well as against the C-terminus of envoplakin. By ELISA, reactivities against envoplakin(1-481), envoplakin(1626-2033), and periplakin(1-324) were found in 25 (80.6%), 25 (80.6%), and 23 (74.2%) PNP sera, and in 1 (1.2%), 3 (3.7%), and 2 (2.5%) control sera, respectively.CONCLUSIONS: The new ELISA based on an N-terminal fragment of envoplakin shows a high diagnostic accuracy to detect circulating autoantibodies in PNP. It is easy to be setup and standardized and can help to differentiate PNP patients from those with pemphigus vulgaris.

U2 - 10.1016/j.cca.2009.08.022

DO - 10.1016/j.cca.2009.08.022

M3 - Journal articles

C2 - 19737550

SN - 0009-8981

VL - 410

SP - 13

EP - 18

JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

IS - 1-2

ER -

Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus

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