Identification of new autoantigens for primary biliary cirrhosis using human proteome microarrays.
Primary biliary cirrhosis (PBC) is a chronic cholestatic liver disease of unknown etiology and is considered to be an autoimmune disease. Autoantibodies are important tools for accurate diagnosis of PBC. Here, we employed serum profiling analysis using a human proteome microarray composed of about 17,000 full-length unique proteins and identified 23 proteins that correlated with PBC. To validate these results, we fabricated a PBC-focused microarray with 21 of these newly identified candidates and nine additional known PBC antigens. By screening the PBC microarrays with additional cohorts of 191 PBC patients and 321 controls (43 autoimmune hepatitis, 55 hepatitis B virus, 31 hepatitis C virus, 48 rheumatoid arthritis, 45 systematic lupus erythematosus, 49 systemic sclerosis, and 50 healthy), six proteins were confirmed as novel PBC autoantigens with high sensitivities and specificities, including hexokinase-1 (isoforms I and II), Kelch-like protein 7, Kelch-like protein 12, zinc finger and BTB domain-containing protein 2, and eukaryotic translation initiation factor 2C, subunit 1. To facilitate clinical diagnosis, we developed ELISA for Kelch-like protein 12 and zinc finger and BTB domain-containing protein 2 and tested large cohorts (297 PBC and 637 control sera) to confirm the sensitivities and specificities observed in the microarray-based assays. In conclusion, our research showed that a strategy using high content protein microarray combined with a smaller but more focused protein microarray can effectively identify and validate novel PBC-specific autoantigens and has the capacity to be translated to clinical diagnosis by means of an ELISA-based method.
Department of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100032, China.
Chao-Jun
CJ
Hu
Guang
G
Song
Wei
W
Huang
Guo-Zhen
GZ
Liu
Chui-Wen
CW
Deng
Hai-Pan
HP
Zeng
Li
L
Wang
Feng-Chun
FC
Zhang
Xuan
X
Zhang
Jun Seop
JS
Jeong
Seth
S
Blackshaw
Li-Zhi
LZ
Jiang
Heng
H
Zhu
Lin
L
Wu
Yong-Zhe
YZ
Li
D000328:Adult; D060565:Argonaute Proteins / Q000276:immunology; D001323:Autoantibodies / Q000097:blood* / Q000276:immunology; D001324:Autoantigens / Q000032:analysis* / Q000276:immunology; D001327:Autoimmune Diseases / Q000175:diagnosis / Q000276:immunology; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology; D039642:Eukaryotic Initiation Factors / Q000276:immunology; D005260:Female; D006593:Hexokinase / Q000032:analysis / Q000276:immunology; D006801:Humans; D008105:Liver Cirrhosis, Biliary / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D008297:Male; D008875:Middle Aged; D040081:Protein Array Analysis; D020543:Proteome / Q000032:analysis* / Q000276:immunology; D012097:Repressor Proteins / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity; D016335:Zinc Fingers / Q000276:immunology
C115848:AGO1 protein, human; D060565:Argonaute Proteins; D001323:Autoantibodies; D001324:Autoantigens; D002352:Carrier Proteins; D039642:Eukaryotic Initiation Factors; C473356:KLHL6 protein, human; D020543:Proteome; D012097:Repressor Proteins; C472074:HK1 protein, human; D006593:Hexokinase