Characterization and use of the ECV304 autoantigenic citrullinome to understand anti-citrullinated protein/peptide autoantibodies in rheumatoid arthritis

Simple item page
dc.contributor.authorde França, Natalia R.
dc.contributor.authorMénard, Henri A.
dc.contributor.authorLora, Maximilien
dc.contributor.authorZhou, Zhijie
dc.contributor.authorRauch, Joyce
dc.contributor.authorHitchon, Carol
dc.contributor.authorAndrade, Luís E. C.
dc.contributor.authorColmegna, Inés
dc.date.accessioned2022-02-01T04:46:00Z
dc.date.issued2022-01-13
dc.date.updated2022-02-01T04:46:00Z
dc.description.abstractAbstract 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.
dc.identifier.citationArthritis Research & Therapy. 2022 Jan 13;24(1):23
dc.identifier.urihttps://doi.org/10.1186/s13075-021-02698-2
dc.identifier.urihttp://hdl.handle.net/1993/36241
dc.language.rfc3066en
dc.rightsopen accessen_US
dc.rights.holderThe Author(s)
dc.titleCharacterization and use of the ECV304 autoantigenic citrullinome to understand anti-citrullinated protein/peptide autoantibodies in rheumatoid arthritis
dc.typeJournal Article
local.author.affiliationRady Faculty of Health Sciencesen_US
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
13075_2021_Article_2698.pdf
Size:
1.33 MB
Format:
Adobe Portable Document Format
Description:
Download
License bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
license.txt
Size:
2.24 KB
Format:
Item-specific license agreed to upon submission
Description:
Download
Collections
University of Manitoba Scholarship
Rady Faculty of Health Sciences Scholarly Works