Detection of anti- periplakin auto-antibodies during idiopathic pulmonary fibrosis Sabine Mignot, Camille Taille, Valerie Besnard, Bruno Cretani, Sylvie CHollet-Marting PII: DOI: Reference:
S0009-8981(14)00135-1 doi: 10.1016/j.cca.2014.03.027 CCA 13462
To appear in:
Clinica Chimica Acta
Received date: Accepted date:
28 February 2014 20 March 2014
Please cite this article as: Mignot Sabine, Taille Camille, Besnard Valerie, Cretani Bruno, CHollet-Marting Sylvie, Detection of anti- periplakin auto-antibodies during idiopathic pulmonary fibrosis, Clinica Chimica Acta (2014), doi: 10.1016/j.cca.2014.03.027
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Letter to the editor
IP
T
Detection of anti- periplakin auto-antibodies during idiopathic pulmonary fibrosis
SC R
Dear Editor,
NU
In their recent publication [1], Muro Y et al described new ELISAs for the diagnosis of paraneoplastic pemphigus, based on the detection of anti-envoplakin and
MA
anti-periplakin antibodies (Abs). The authors included in their study 15 patients with
D
idiopathic pulmonary fibrosis (IPF), and concluded that none of these patients were
TE
positive for anti-periplakin Abs. As mentioned by the authors, their results were not
CE P
concordant with those of our group as we detected anti-periplakin Abs in 40% IPF patients [2]. We comment on this work and highlight some limitations in the validation of
AC
the anti-periplakin ELISA, which, we believe, preclude any firm conclusion. We consider that the sensitivity of the anti-periplakin and anti-envoplakin assays
developed by Muro Y et al. was not adequately assessed. The cut-off level of the assay was set to 3 SDs above the mean value obtained from 20 healthy control sera. Only one serum from a patient with paraneoplastic pemphigus was used as a positive control. Additional sera are required to validate the sensitivity of the assay. It would have been important to determine the cut off value differentiating at least 50 positive and negative
ACCEPTED MANUSCRIPT
sera, and to perform receiver-operating-characteristic (ROC) analysis, as in previous
SC R
detection of anti-periplakin or anti-envoplakin antibodies.
IP
T
studies [3,4]. As long as this work is not done, the ELISAs cannot be used for routine
These technical limitations have major implications. In particular, in one patient
NU
with IPF, 2 bands around 200 kDa were detected by immunoblot with the human
MA
epidermal extract; however, the authors excluded the presence of anti-envoplakin or anti-periplakin on the basis of the negative results of both ELISAs. Since the sensitivity
D
of the assay was not established, we assume that the authors cannot exclude the presence
TE
of anti-periplakin Abs on this sole basis.
CE P
In our routine procedure, all sera are simultaneously evaluated by immunoblot on a human placental extract (native protein) and on the recombinant periplakin protein.
AC
The 200kDa band is of lower intensity in IPF sera than in paraneoplastic pemphigus; however, sera with a 200kDa band on the human placental extract are unambiguously positive using the recombinant protein. The presence of anti-periplakin Abs in IPF patients has now been confirmed by our group, studying >100 patients [5]. Analysis of the prognostic value and clinical relevance of the anti-periplakin Abs in IPF patients is currently under investigation. Sabine Mignot
ACCEPTED MANUSCRIPT
SC R
IP
T
Camille Taille Valerie Besnard Bruno Cretani
Sylvie CHollet-Marting
NU
Bichat Hospital, Immunology, 46 rue Henri Huchard, PARIS, 75018, France,
MA
33140258531, [email protected] References
D
[1] Y. Muro, K. Sugiura, A. Shiraki, N. Ishii, T. Hashimoto, M. Akiyama, Detection of
TE
autoantibodies to periplakin and envoplakin in paraneoplastic pemphigus but not
CE P
idiopathic pulmonary fibrosis using full-length recombinant proteins, Clin. Chim. Acta 429 (2014) 14-17.
AC
[2] C. Taillé, S. Grootenboer-Mignot, C. Boursier, M.P. Debray, J. Fagart, A.C. Steven, L. Barrientos, A. Mailleux, N. Cigna, F. Tubach, J. Marchal-Sommé, P. Soler, S. Chollet-Martin, B. Crestani, Identification of periplakin as a new target for autoreactivity in Idiopathic Pulmonary Fibrosis,. Am. J. Respir. Crit. Care Med. 183 (2011) 759-66. [3] C. Probst, W. Schlumberger, W. Stocker, A. Recke, E. Schmidt, T. Hashimoto, X.J. Zhu, D. Zillikens, L. Komorowski, Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus, Clin.
ACCEPTED MANUSCRIPT
Chim. Acta 410 (2009) 13-18.
IP
T
[4] Y. Huang, J. Li, X. Zhu, Detection of anti-envoplakin and anti-periplakin
SC R
autoantibodies by ELISA in patients with paraneoplastic pemphigus, Arch. Dermatol. Res. 30 (2009) 703-709.
NU
[5] S. Mignot, C. Taillé, S. Ly Ka Su, C. Champagnat, J. Fagart, V. Besnard, V. Descamps,
MA
B.Crestani, S. Chollet-Martin, Presence of periplakin auto-antibodies during idiopathic pulmonary fibrosis, GREMI Meeting, “The underestimated role of epithelium in
AC
CE P
TE
D
inflammation”, April, 12th, 2013 – Pasteur Institute, Paris, France.
S0009-8981(14)00135-1 doi: 10.1016/j.cca.2014.03.027 CCA 13462
To appear in:
Clinica Chimica Acta
Received date: Accepted date:
28 February 2014 20 March 2014
Please cite this article as: Mignot Sabine, Taille Camille, Besnard Valerie, Cretani Bruno, CHollet-Marting Sylvie, Detection of anti- periplakin auto-antibodies during idiopathic pulmonary fibrosis, Clinica Chimica Acta (2014), doi: 10.1016/j.cca.2014.03.027
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Letter to the editor
IP
T
Detection of anti- periplakin auto-antibodies during idiopathic pulmonary fibrosis
SC R
Dear Editor,
NU
In their recent publication [1], Muro Y et al described new ELISAs for the diagnosis of paraneoplastic pemphigus, based on the detection of anti-envoplakin and
MA
anti-periplakin antibodies (Abs). The authors included in their study 15 patients with
D
idiopathic pulmonary fibrosis (IPF), and concluded that none of these patients were
TE
positive for anti-periplakin Abs. As mentioned by the authors, their results were not
CE P
concordant with those of our group as we detected anti-periplakin Abs in 40% IPF patients [2]. We comment on this work and highlight some limitations in the validation of
AC
the anti-periplakin ELISA, which, we believe, preclude any firm conclusion. We consider that the sensitivity of the anti-periplakin and anti-envoplakin assays
developed by Muro Y et al. was not adequately assessed. The cut-off level of the assay was set to 3 SDs above the mean value obtained from 20 healthy control sera. Only one serum from a patient with paraneoplastic pemphigus was used as a positive control. Additional sera are required to validate the sensitivity of the assay. It would have been important to determine the cut off value differentiating at least 50 positive and negative
ACCEPTED MANUSCRIPT
sera, and to perform receiver-operating-characteristic (ROC) analysis, as in previous
SC R
detection of anti-periplakin or anti-envoplakin antibodies.
IP
T
studies [3,4]. As long as this work is not done, the ELISAs cannot be used for routine
These technical limitations have major implications. In particular, in one patient
NU
with IPF, 2 bands around 200 kDa were detected by immunoblot with the human
MA
epidermal extract; however, the authors excluded the presence of anti-envoplakin or anti-periplakin on the basis of the negative results of both ELISAs. Since the sensitivity
D
of the assay was not established, we assume that the authors cannot exclude the presence
TE
of anti-periplakin Abs on this sole basis.
CE P
In our routine procedure, all sera are simultaneously evaluated by immunoblot on a human placental extract (native protein) and on the recombinant periplakin protein.
AC
The 200kDa band is of lower intensity in IPF sera than in paraneoplastic pemphigus; however, sera with a 200kDa band on the human placental extract are unambiguously positive using the recombinant protein. The presence of anti-periplakin Abs in IPF patients has now been confirmed by our group, studying >100 patients [5]. Analysis of the prognostic value and clinical relevance of the anti-periplakin Abs in IPF patients is currently under investigation. Sabine Mignot
ACCEPTED MANUSCRIPT
SC R
IP
T
Camille Taille Valerie Besnard Bruno Cretani
Sylvie CHollet-Marting
NU
Bichat Hospital, Immunology, 46 rue Henri Huchard, PARIS, 75018, France,
MA
33140258531, [email protected] References
D
[1] Y. Muro, K. Sugiura, A. Shiraki, N. Ishii, T. Hashimoto, M. Akiyama, Detection of
TE
autoantibodies to periplakin and envoplakin in paraneoplastic pemphigus but not
CE P
idiopathic pulmonary fibrosis using full-length recombinant proteins, Clin. Chim. Acta 429 (2014) 14-17.
AC
[2] C. Taillé, S. Grootenboer-Mignot, C. Boursier, M.P. Debray, J. Fagart, A.C. Steven, L. Barrientos, A. Mailleux, N. Cigna, F. Tubach, J. Marchal-Sommé, P. Soler, S. Chollet-Martin, B. Crestani, Identification of periplakin as a new target for autoreactivity in Idiopathic Pulmonary Fibrosis,. Am. J. Respir. Crit. Care Med. 183 (2011) 759-66. [3] C. Probst, W. Schlumberger, W. Stocker, A. Recke, E. Schmidt, T. Hashimoto, X.J. Zhu, D. Zillikens, L. Komorowski, Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus, Clin.
ACCEPTED MANUSCRIPT
Chim. Acta 410 (2009) 13-18.
IP
T
[4] Y. Huang, J. Li, X. Zhu, Detection of anti-envoplakin and anti-periplakin
SC R
autoantibodies by ELISA in patients with paraneoplastic pemphigus, Arch. Dermatol. Res. 30 (2009) 703-709.
NU
[5] S. Mignot, C. Taillé, S. Ly Ka Su, C. Champagnat, J. Fagart, V. Besnard, V. Descamps,
MA
B.Crestani, S. Chollet-Martin, Presence of periplakin auto-antibodies during idiopathic pulmonary fibrosis, GREMI Meeting, “The underestimated role of epithelium in
AC
CE P
TE
D
inflammation”, April, 12th, 2013 – Pasteur Institute, Paris, France.
