An immuno-DOT diagnostic assay for autoimmune nodopathy

Alexandre Jentzer; Guillaume Taieb; Jérémie El Bechir; Thierry Vincent; Jérôme Joël Devaux

doi:10.1515/cclm-2024-0510

Article

An immuno-DOT diagnostic assay for autoimmune nodopathy

Alexandre Jentzer , Guillaume Taieb , Jérémie El Bechir , Thierry Vincent and Jérôme Joël Devaux

Published/Copyright: June 12, 2024

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From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 63 Issue 2

Abstract

Objectives

Autoimmune nodopathy (AN) is a life-threatening peripheral neuropathy mediated by four autoantibodies targeting axoglial cell adhesion molecules at the nodes of Ranvier: Neurofascin-155 (Nfasc155), PanNeurofascin (PanNfasc), Contactin-1 (CNTN1), and Contactin-associated protein 1 (CASPR1). Antibody detection is a strong biomarker for AN diagnosis and treatment monitoring. The aim of this study was to develop an immuno-dot assay (immuno-DOT) compatible with routine implementation in medical laboratories.

Methods

This new approach was compared to standard techniques: indirect immunofluorescence assay, cell-based assay, and ELISA. Sensitivities (Se) and specificities (Sp) were calculated on a cohort composed of 58 patients diagnosed with AN, 50 seronegative patients with chronic inflammatory demyelinating polyradiculoneuropathy, 20 healthy controls, 30 patients with Guillain-Barré syndrome, 20 with monoclonal gammopathy and 20 with Charcot-Marie-Tooth disease. The patients were diagnosed with AN based on compatible electro-clinical arguments and at least two positive standard techniques.

Results

Immuno-DOT sensitivities and specificities were Se=91 %, Sp=97 % for anti-Nfasc155; Se=80 %, Sp=94 % for anti-PanNfasc; Se=93 %, Sp=98 % for anti-CNTN1; and Se=87 %, Sp=94 % for anti-CASPR1. Immuno-DOT allowed the diagnosis within 3 h and the accurate follow-up of the immune reactivity and isotype, and dot intensity correlated with antibody titers following treatments. A longitudinal study indicated that immuno-DOT yielded reliable results even after six months of storage at −20 °C.

Conclusions

The diagnostic performance of immuno-DOT was satisfactory and compatible with routine implementation in medical laboratories.

Keywords: autoimmune nodopathy; chronic inflammatory demyelinating polyneuropathy (CIDP); IgG4; immuno-DOT; NF155

Corresponding authors: Alexandre Jentzer, Institut de Génomique Fonctionnelle, Université de Montpellier, CNRS, INSERM, Montpellier, France; Department of Immunology, Saint Eloi University Hospital Center, Montpellier University, Montpellier, France; and Laboratoire d’Immunologie, CHU Montpellier, 80 avenue Augustin Fliche, 34295, Montpellier Cedex 5, France, E-mail: a-jentzer@chu-montpellier.fr; and Jérôme Devaux, Institut de Génomique Fonctionnelle, Université de Montpellier, CNRS, INSERM, Montpellier, France, E-mail: jerome.devaux@igf.cnrs.fr

Funding source: Agence Nationale pour la Recherche

Award Identifier / Grant number: NECCIN

Funding source: French Muscular Dystrophy Association

Award Identifier / Grant number: grant#23593

Research ethics: All the subjects provided written informed consent. The study was approved by the Ethics Committee of Montpellier University Hospital (IRB-MTP-2020-01-20200339). All animal experiments were in line with the European community’s guiding principles on the care and use of animals (2010/63/EU) and were approved by the local Ethical Committee and by the “ministére de l’éducation nationale de l’enseignement supérieur et de la recherche” (APAFIS#3847-2016012610089856v5).
Informed consent: Informed consent was obtained from all individuals included in this study, or their legal guardians or wards.
Author contributions: Alexandre Jentzer Jérôme Joël Devaux contributed to conception, design, acquisition of data, analysis and interpretation. All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Competing interests: Dr. Devaux received a research grant from CSL Behring. The other authors have no financial conflicts of interest.
Research funding: This work was supported by the Agence Nationale pour la Recherche (NECCIN; GT and JJD), from the Association Française contre les Myopathies (grant#23593; GT and JJD) and ArgenX.
Data availability: The raw data can be obtained on request from the corresponding author.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2024-0510).

Received: 2023-11-10

Accepted: 2024-05-30

Published Online: 2024-06-12

Published in Print: 2025-01-29

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Supplementary Material

Articles in the same Issue

https://doi.org/10.1515/cclm-2024-0510

Keywords for this article

autoimmune nodopathy; chronic inflammatory demyelinating polyneuropathy (CIDP); IgG4; immuno-DOT; NF155