Proteomic analysis of secreted proteins in early rheumatoid arthritis: anti-citrulline autoreactivity is associated with up regulation of proinflammatory cytokines.
OBJECTIVE
To identify peripheral blood autoantibody and cytokine profiles that characterise clinically relevant subgroups of patients with early rheumatoid arthritis using arthritis antigen microarrays and a multiplex cytokine assay.


METHODS
Serum samples from 56 patients with a diagnosis of rheumatoid arthritis of <6 months' duration were tested. Cytokine profiles were also determined in samples from patients with psoriatic arthritis (PsA) and ankylosing spondylitis (n = 21), and from healthy individuals (n = 19). Data were analysed using Kruskal-Wallis test with Dunn's adjustment for multiple comparisons, linear correlation tests, significance analysis of microarrays (SAM) and hierarchical clustering software.


RESULTS
Distinct antibody profiles were associated with subgroups of patients who exhibited high serum levels of tumour necrosis factor (TNF)alpha, interleukin (IL)1beta, IL6, IL13, IL15 and granulocyte macrophage colony-stimulating factor. Significantly increased autoantibody reactivity against citrullinated epitopes was observed in patients within the cytokine "high" subgroup. Increased levels of TNFalpha, IL1alpha, IL12p40 and IL13, and the chemokines eotaxin/CCL11, monocyte chemoattractant protein-1 and interferon-inducible protein 10, were present in early rheumatoid arthritis as compared with controls (p<0.001). Chemokines showed some of the most impressive differences. Only IL8/CXCL8 concentrations were higher in patients with PsA/ankylosing spondylitis (p = 0.02).


CONCLUSIONS
Increased blood levels of proinflammatory cytokines are associated with autoantibody targeting of citrullinated antigens and surrogate markers of disease activity in patients with early rheumatoid arthritis. Proteomic analysis of serum autoantibodies, cytokines and chemokines enables stratification of patients with early rheumatoid arthritis into molecular subgroups.
Hueber
Wolfgang
W
Division of Immunology and Rheumatology, Department of Medicine, Stanford University School of Medicine, and Palo Alto VA Health Care System, MC 154R, 3801 Miranda Avenue, Palo Alto, CA 94304, USA. whueber@stanford.edu
Tomooka
Beren H
BH
Zhao
Xiaoyan
X
Kidd
Brian A
BA
Drijfhout
Jan W
JW
Fries
James F
JF
van Venrooij
Walther J
WJ
Metzger
Allan L
AL
Genovese
Mark C
MC
Robinson
William H
WH
D000328:Adult; D000368:Aged; D015535:Arthritis, Psoriatic / Q000276:immunology; D001172:Arthritis, Rheumatoid / Q000276:immunology*; D001323:Autoantibodies / Q000097:blood*; D001324:Autoantigens / Q000276:immunology; D015415:Biomarkers / Q000097:blood; D018925:Chemokines / Q000097:blood; D002956:Citrulline / Q000276:immunology*; D016207:Cytokines / Q000096:biosynthesis* / Q000097:blood; D005260:Female; D006801:Humans; D018836:Inflammation Mediators / Q000097:blood*; D008297:Male; D008875:Middle Aged; D040081:Protein Array Analysis / Q000379:methods; D040901:Proteomics; D013167:Spondylitis, Ankylosing / Q000276:immunology; D015854:Up-Regulation
D001323:Autoantibodies; D001324:Autoantigens; D015415:Biomarkers; D018925:Chemokines; D016207:Cytokines; D018836:Inflammation Mediators; D002956:Citrulline