title: Comparative analysis of autoantibodies targeting peptidylarginine deiminase type 4, mutated citrullinated vimentin and cyclic citrullinated peptides in rheumatoid arthritis: associations with cytokine profiles, clinical and genetic features.
abstract: Antibodies against cyclic citrullinated peptides (anti-CCP) are widely used for diagnosis of rheumatoid arthritis (RA). We performed a comparative analysis of antibodies targeting the citrullinating enzyme peptidylarginine deiminase type 4 (anti-PAD4) and mutated citrullinated vimentin (anti-MCV) with anti-CCP autoantibodies in RA patients and examined their relationships with clinical parameters, cytokine profiles and the PADI4 gene. Autoantibodies were examined by enzyme-linked immunosorbent assay (ELISA) in sera of 170 RA patients and 103 controls. Cytokine profiles were measured using a multiplex system. PADI4 polymorphisms (89 G > A, 90 T > C and 92 G > C) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Anti-PAD4, anti-MCV and anti-CCP autoantibodies were detected in 24, 61 and 74% of RA patients, respectively. Positive correlations were observed between anti-PAD4 and disease duration; anti-CCP and erythrocyte sedimentation rate (ESR); anti-MCV and ESR and C-reactive protein. Anti-MCV antibodies were associated with high disease activity score 28 (DAS-28) in early RA. Concentrations of T helper type 1 (Th1) [tumour necrosis factor (TNF)-α, interleukin (IL)-12, IL-2, IL-1β], Th2 (IL-4, IL-6, IL-10, IL-13) and Th17 (IL-17) cytokines were higher in RA than in controls. Th2 and, to a lesser extent, Th1-related cytokines, showed positive correlations with anti-MCV and anti-CCP. The GTG haplotype in PADI4 was associated with anti-CCP and anti-MCV, but not anti-PAD4 antibodies. In conclusion, anti-PAD4 antibodies are detected mainly in established RA, which is in contrast to the early detection of antibodies against citrullinated peptide/proteins (ACPAs). Among autoantibodies, anti-MCV appear to perform better as markers of disease activity. Furthermore, anti-CCP and anti-MCV are associated genetically with the citrullinating enzyme PAD4 and are related strongly to Th1 and Th2 cytokines, suggesting a feed-forward loop between cytokines and ACPA production.
authors.affiliation: Research Institute in Biomedical Sciences, Department of Medical Clinics, CUCS, Universidad De Guadalajara, Guadalajara, Jalisco.
authors.forename: Z
authors.initials: Z
authors.lastname: Reyes-Castillo
authors.affiliation: Research Institute in Biomedical Sciences, Department of Medical Clinics, CUCS, Universidad De Guadalajara, Guadalajara, Jalisco.
authors.forename: C A
authors.initials: CA
authors.lastname: Palafox-Sánchez
authors.affiliation: Academic Department of Chemistry and Biologic Sciences, Universidad Autonoma De Guerrero, Chilpancingo, Guerrero.
authors.forename: I
authors.initials: I
authors.lastname: Parra-Rojas
authors.affiliation: Division of Rheumatology, Hospital Civil De Guadalajara 'Fray Antonio Alcalde', Guadalajara, Jalisco.
authors.forename: G E
authors.initials: GE
authors.lastname: Martínez-Bonilla
authors.affiliation: Institute of Chronic and Degenerative Diseases, Department of Molecular Biology and Genomics, CUCS, Universidad De Guadalajara, Guadalajara, Jalisco, Mexico.
authors.forename: S
authors.initials: S
authors.lastname: del Toro-Arreola
authors.affiliation: Research Institute in Biomedical Sciences, Department of Medical Clinics, CUCS, Universidad De Guadalajara, Guadalajara, Jalisco.
authors.forename: M G
authors.initials: MG
authors.lastname: Ramírez-Dueñas
authors.affiliation: Research Institute in Biomedical Sciences, Department of Medical Clinics, CUCS, Universidad De Guadalajara, Guadalajara, Jalisco.
authors.forename: G
authors.initials: G
authors.lastname: Ocampo-Bermudes
authors.affiliation: Research Institute in Biomedical Sciences, Department of Medical Clinics, CUCS, Universidad De Guadalajara, Guadalajara, Jalisco.
authors.forename: José F
authors.initials: JF
authors.lastname: Muñoz-Valle
mesh_terms: D000328:Adult; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000235:genetics / Q000276:immunology*; D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D001799:Blood Sedimentation; D002956:Citrulline / Q000737:chemistry; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D006801:Humans; D006867:Hydrolases / Q000235:genetics / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D050505:Mutant Proteins / Q000276:immunology; D010456:Peptides, Cyclic / Q000276:immunology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases; D012720:Severity of Illness Index; D018417:Th1 Cells / Q000276:immunology / Q000378:metabolism; D058504:Th17 Cells / Q000276:immunology / Q000378:metabolism; D018418:Th2 Cells / Q000276:immunology / Q000378:metabolism; D014746:Vimentin / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D001323:Autoantibodies; D016207:Cytokines; D050505:Mutant Proteins; D010456:Peptides, Cyclic; D014746:Vimentin; C487763:cyclic citrullinated peptide; D002956:Citrulline; D006867:Hydrolases; C000629235:PADI4 protein, human; D000080002:Protein-Arginine Deiminase Type 4; D000076342:Protein-Arginine Deiminases
keywords: ACPA; PADI4; citrullination, disease activity; rheumatoid arthritis