Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodies
Autoantibodies (Abs) are biomarkers for many disease conditions and are increasingly used to facilitate diagnosis and treatment decisions. To guarantee high sensitivity and specificity, the choice of their detection method is crucial. Via cell-based assays, we recently found 21 patients with neurolo...
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.972161/full |
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| author | Christian P. Moritz Christian P. Moritz Christian P. Moritz Le-Duy Do Le-Duy Do Yannick Tholance Yannick Tholance Yannick Tholance Pierre-Baptiste Vallayer Véronique Rogemond Véronique Rogemond Bastien Joubert Bastien Joubert Karine Ferraud Karine Ferraud Coralie La Marca Coralie La Marca Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jérôme Honnorat Jérôme Honnorat Jérôme Honnorat Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine |
| author_facet | Christian P. Moritz Christian P. Moritz Christian P. Moritz Le-Duy Do Le-Duy Do Yannick Tholance Yannick Tholance Yannick Tholance Pierre-Baptiste Vallayer Véronique Rogemond Véronique Rogemond Bastien Joubert Bastien Joubert Karine Ferraud Karine Ferraud Coralie La Marca Coralie La Marca Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jérôme Honnorat Jérôme Honnorat Jérôme Honnorat Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine |
| author_sort | Christian P. Moritz |
| collection | DOAJ |
| description | Autoantibodies (Abs) are biomarkers for many disease conditions and are increasingly used to facilitate diagnosis and treatment decisions. To guarantee high sensitivity and specificity, the choice of their detection method is crucial. Via cell-based assays, we recently found 21 patients with neurological diseases positive for antibodies against argonaute (AGO), 10 of which having a neuropathy (NP). Here, we established a simple and conformation-sensitive ELISA with the aim to distinguish between AGO1 Abs against conformational epitopes and non-conformational epitopes and to reveal further characteristics of AGO1 antibodies in NP and autoimmune disease (AID). In a retrospective multicenter case/control and observational study, we tested 434 patients with NP, 274 disease controls with AID, and 116 healthy controls (HC) for AGO1 Abs via conformation-stabilizing ELISA. Seropositive patients were also tested for conformation-specificity via comparative denaturing/stabilizing ELISA (CODES-ELISA), CBA positivity, AGO1 titers and IgG subclasses, and AGO2 reactivity. These parameters were statistically compared among different epitope-specific patient groups. We found Abs in 44 patients, including 28/434 (6.5%) NP, 16/274 (5.8%) AID, and 0/116 (0%) HC. Serum reactivity was consistently higher for AGO1 than AGO2. Globally among the 44 AGO1 Abs-positive patients, 42 were also tested in CBA for AGO1 Abs positivity and 15 (35.7%) were positive. Furthermore, 43 were tested for conformation-specificity and 32 (74.4%) bound a conformational epitope. Among the subgroups of highly positive patients (ELISA z-score >14) with sera binding conformational epitopes (n=23), 14 patient sera were also CBA positive and 9 bound a second conformational but CBA-inaccessible epitope. A third, non-conformational epitope was bound by 11/43 (15.6%). Among the epitope-specific patient subgroups, we found significant differences regarding the Abs titers, IgG subclass, and AGO2 reactivity. When comparing AGO1 Abs-positive NP versus AID patients, we found the conformation-specific and CBA inaccessible epitope significantly more frequently in AID patients. We conclude that 1) conformational ELISA was more sensitive than CBA in detecting AGO1 Abs, 2) serum reactivity is higher for AGO1 than for AGO2 at least for NP patients, 3) AGO1 Abs might be a marker-of-interest in 6.5% of NP patients, 4) distinguishing epitopes might help finding different patient subgroups. |
| first_indexed | 2024-04-13T18:36:27Z |
| format | Article |
| id | doaj.art-b877a0ad925047ba9e90340284c81c20 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-13T18:36:27Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-b877a0ad925047ba9e90340284c81c202022-12-22T02:34:53ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-10-011310.3389/fimmu.2022.972161972161Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodiesChristian P. Moritz0Christian P. Moritz1Christian P. Moritz2Le-Duy Do3Le-Duy Do4Yannick Tholance5Yannick Tholance6Yannick Tholance7Pierre-Baptiste Vallayer8Véronique Rogemond9Véronique Rogemond10Bastien Joubert11Bastien Joubert12Karine Ferraud13Karine Ferraud14Coralie La Marca15Coralie La Marca16Jean-Philippe Camdessanché17Jean-Philippe Camdessanché18Jean-Philippe Camdessanché19Jean-Philippe Camdessanché20Jean-Philippe Camdessanché21Jérôme Honnorat22Jérôme Honnorat23Jérôme Honnorat24Jean-Christophe Antoine25Jean-Christophe Antoine26Jean-Christophe Antoine27Jean-Christophe Antoine28Jean-Christophe Antoine29Department of Neurology, University hospital of Saint-Etienne, Saint-Etienne, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceINMG/Melys team, University Jean Monnet, Saint-Étienne, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceFrench Reference Center on Paraneoplastic Neurological Syndromes and Autoimmune Encephalitis, Hospices Civils de Lyon, Bron, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceINMG/Melys team, University Jean Monnet, Saint-Étienne, FranceDepartment of Biochemistry, University Hospital of Saint-Etienne, Saint-Etienne, FranceDepartment of Neurology, University hospital of Saint-Etienne, Saint-Etienne, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceFrench Reference Center on Paraneoplastic Neurological Syndromes and Autoimmune Encephalitis, Hospices Civils de Lyon, Bron, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceFrench Reference Center on Paraneoplastic Neurological Syndromes and Autoimmune Encephalitis, Hospices Civils de Lyon, Bron, FranceDepartment of Neurology, University hospital of Saint-Etienne, Saint-Etienne, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceDepartment of Biochemistry, University Hospital of Saint-Etienne, Saint-Etienne, FranceDepartment of Neurology, University hospital of Saint-Etienne, Saint-Etienne, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceINMG/Melys team, University Jean Monnet, Saint-Étienne, FranceFrench Reference Center on Paraneoplastic Neurological Syndromes and Autoimmune Encephalitis, Hospices Civils de Lyon, Bron, FranceEuropean Reference Network for Rare Neuromuscular Diseases, Paris, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceFrench Reference Center on Paraneoplastic Neurological Syndromes and Autoimmune Encephalitis, Hospices Civils de Lyon, Bron, FranceEuropean Reference Network for Rare Neuromuscular Diseases, Paris, FranceDepartment of Neurology, University hospital of Saint-Etienne, Saint-Etienne, FranceSynaptopathies and autoantibodies (SynatAc) team, Mechanisms In Integrated Life Sciences (MELIS) Laboratory, Institute NeuroMyoGène (INMG), INSERM U1314/CNRS UMR 5284, Universités de Lyon, Université Claude Bernard Lyon 1, Lyon, FranceINMG/Melys team, University Jean Monnet, Saint-Étienne, FranceFrench Reference Center on Paraneoplastic Neurological Syndromes and Autoimmune Encephalitis, Hospices Civils de Lyon, Bron, FranceEuropean Reference Network for Rare Neuromuscular Diseases, Paris, FranceAutoantibodies (Abs) are biomarkers for many disease conditions and are increasingly used to facilitate diagnosis and treatment decisions. To guarantee high sensitivity and specificity, the choice of their detection method is crucial. Via cell-based assays, we recently found 21 patients with neurological diseases positive for antibodies against argonaute (AGO), 10 of which having a neuropathy (NP). Here, we established a simple and conformation-sensitive ELISA with the aim to distinguish between AGO1 Abs against conformational epitopes and non-conformational epitopes and to reveal further characteristics of AGO1 antibodies in NP and autoimmune disease (AID). In a retrospective multicenter case/control and observational study, we tested 434 patients with NP, 274 disease controls with AID, and 116 healthy controls (HC) for AGO1 Abs via conformation-stabilizing ELISA. Seropositive patients were also tested for conformation-specificity via comparative denaturing/stabilizing ELISA (CODES-ELISA), CBA positivity, AGO1 titers and IgG subclasses, and AGO2 reactivity. These parameters were statistically compared among different epitope-specific patient groups. We found Abs in 44 patients, including 28/434 (6.5%) NP, 16/274 (5.8%) AID, and 0/116 (0%) HC. Serum reactivity was consistently higher for AGO1 than AGO2. Globally among the 44 AGO1 Abs-positive patients, 42 were also tested in CBA for AGO1 Abs positivity and 15 (35.7%) were positive. Furthermore, 43 were tested for conformation-specificity and 32 (74.4%) bound a conformational epitope. Among the subgroups of highly positive patients (ELISA z-score >14) with sera binding conformational epitopes (n=23), 14 patient sera were also CBA positive and 9 bound a second conformational but CBA-inaccessible epitope. A third, non-conformational epitope was bound by 11/43 (15.6%). Among the epitope-specific patient subgroups, we found significant differences regarding the Abs titers, IgG subclass, and AGO2 reactivity. When comparing AGO1 Abs-positive NP versus AID patients, we found the conformation-specific and CBA inaccessible epitope significantly more frequently in AID patients. We conclude that 1) conformational ELISA was more sensitive than CBA in detecting AGO1 Abs, 2) serum reactivity is higher for AGO1 than for AGO2 at least for NP patients, 3) AGO1 Abs might be a marker-of-interest in 6.5% of NP patients, 4) distinguishing epitopes might help finding different patient subgroups.https://www.frontiersin.org/articles/10.3389/fimmu.2022.972161/fullargonaute antibodiessensory neuronopathyganglionopathieAnti-Su antibodiesautoimmune neuropathyconformation-stabilizing ELISA |
| spellingShingle | Christian P. Moritz Christian P. Moritz Christian P. Moritz Le-Duy Do Le-Duy Do Yannick Tholance Yannick Tholance Yannick Tholance Pierre-Baptiste Vallayer Véronique Rogemond Véronique Rogemond Bastien Joubert Bastien Joubert Karine Ferraud Karine Ferraud Coralie La Marca Coralie La Marca Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jean-Philippe Camdessanché Jérôme Honnorat Jérôme Honnorat Jérôme Honnorat Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine Jean-Christophe Antoine Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodies Frontiers in Immunology argonaute antibodies sensory neuronopathy ganglionopathie Anti-Su antibodies autoimmune neuropathy conformation-stabilizing ELISA |
| title | Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodies |
| title_full | Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodies |
| title_fullStr | Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodies |
| title_full_unstemmed | Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodies |
| title_short | Conformation-stabilizing ELISA and cell-based assays reveal patient subgroups targeting three different epitopes of AGO1 antibodies |
| title_sort | conformation stabilizing elisa and cell based assays reveal patient subgroups targeting three different epitopes of ago1 antibodies |
| topic | argonaute antibodies sensory neuronopathy ganglionopathie Anti-Su antibodies autoimmune neuropathy conformation-stabilizing ELISA |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.972161/full |
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