Bibliographic Details
| Title: |
Characterization and use of the ECV304 autoantigenic citrullinome to understand anti-citrullinated protein/peptide autoantibodies in rheumatoid arthritis |
| Authors: |
Natalia Regine de França, Henri André Ménard, Maximilien Lora, Zhijie Zhou, Joyce Rauch, Carol Hitchon, Luís Eduardo Coelho Andrade, Inés Colmegna |
| Source: |
Arthritis Research & Therapy, Vol 24, Iss 1, Pp 1-11 (2022) |
| Publisher Information: |
BMC, 2022. |
| Publication Year: |
2022 |
| Collection: |
LCC:Diseases of the musculoskeletal system |
| Subject Terms: |
ECV304, Autocitrullinome, Peptidylarginine deiminases, ACPAs, Rheumatoid arthritis, Diseases of the musculoskeletal system, RC925-935 |
| More Details: |
Abstract Background Anti-citrullinated protein antibodies (ACPAs) are highly specific for rheumatoid arthritis (RA). In vivo, ACPAs target peptidyl-citrulline epitopes (cit-) in a variety of proteins (cit-prot-ACPAs) and derived peptides (cit-pept-ACPAs) generated via the peptidylarginine deiminase (PAD) isoenzymes. We aimed to identify a cell line with self-citrullination capacity, to describe its autoantigenic citrullinome, and to test it as a source of autocitrullinated proteins and peptides. Methods Human cell lines were screened for cit-proteins by Western blot. PAD isoenzymes were identified by RT-PCR. Autocitrullination of ECV304 was optimized, and the ECV304 autocitrullinomes immunoprecipitated by sera from three RA patients were characterized by mass spectrometry. Cit-pept-ACPAs were detected using anti-CCP2 ELISA and cit-prot-ACPAs, by an auto-cit-prot-ECV304 ELISA. Sera from 177 RA patients, 59 non-RA rheumatic disease patients and 25 non-disease controls were tested. Results Of the seven cell lines studied, only ECV304 simultaneously overexpressed PAD2 and PAD3 and its extracts reproducibly autocitrullinated self and non-self-proteins. Proteomic analysis of the cit-ECV304 products immunoprecipitated by RA sera, identified novel cit-targets: calreticulin, profilin 1, vinculin, new 14–3-3 protein family members, chaperones, and mitochondrial enzymes. The auto-cit-prot-ECV304 ELISA had a sensitivity of 50% and a specificity of 95% for RA diagnosis. Conclusions ECV304 cells overexpress two of the PAD isoenzymes capable of citrullinating self-proteins. These autocitrullinated cells constitute a basic and clinical research tool that enable the detection of cit-prot-ACPAs with high diagnostic specificity and allow the identification of the specific cit-proteins targeted by individual RA sera. |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
1478-6362 |
| Relation: |
https://doaj.org/toc/1478-6362 |
| DOI: |
10.1186/s13075-021-02698-2 |
| Access URL: |
https://doaj.org/article/fa4f26124b0349d6892f18d4dfcd9a1b |
| Accession Number: |
edsdoj.fa4f26124b0349d6892f18d4dfcd9a1b |
| Database: |
Directory of Open Access Journals |
