Autoantibody profiling of Chinese patients with autoimmune hepatitis using immunoproteomic analysis.
In the present study, immunoproteomic analysis was utilized to systemically characterize global autoantibody profiles in autoimmune hepatitis (AIH). Sera from 21 patients with AIH and 15 healthy controls were analyzed for the antibody reactivity against the protein antigens of HepG2, a human hepatoma cell line. The lysates of HepG2 cells were separated by two-dimensional electrophoresis and then immunoblotted with each serum sample. Matrix-assisted laser desorption/ionization mass spectrometry or/and nanoelectrospray ionization MS/MS were then used to identify antigens, among which a bifunctional enzyme in mitochondrial, fumarate hydratase (FH), was further analyzed by ELISA using recombinant FH as a coating antigen. A total of 18 immunoreactive spots were identified as 13 proteins, 8 of which have not been reported in AIH. Immune reactivity to FH was detected in 66.67% of patients with AIH, 19.35% of patients with primary biliary cirrhosis (PBC), 12.31% of patients with chronic hepatitis B (CHB), 6.35% of patients with chronic hepatitis C (CHC), 11.32% of patients with systemic lupus erythematosus (SLE), and 3.57% of normal individuals. The differences of prevalence between AIH patients and healthy controls as well as other diseases were of statistical significance (P<0.001). These data demonstrate the serological heterogeneity in AIH and suggest the diversity of the mechanisms underlying AIH. FH, recognized mainly in AIH rather than in viral hepatitis and other autoimmune diseases, may have utility in improved diagnosis of AIH.
Xia
Qing
Q
Department of Molecular Immunology, Institute of Basic Medical Sciences, Taiping Road 27, Beijing 100850, China.
Lu
Feng
F
Yan
Hui-Ping
HP
Wang
Hong-Xia
HX
Feng
Xia
X
Zhao
Yan
Y
Liu
Bing-Yv
BY
Wang
Jie
J
Li
Ping
P
Xue
Yan
Y
Hu
Mei-Ru
MR
Qian
Lu
L
Guo
Ning
N
Yang
Song-Cheng
SC
Li
Ming-Yuan
MY
Ma
Yuan-Fang
YF
Li
Bo-An
BA
Zhang
Xue-Min
XM
Shen
Bei-Fen
BF
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D000818:Animals; D044466:Asian People; D001323:Autoantibodies / Q000032:analysis* / Q000235:genetics; D001324:Autoantigens / Q000276:immunology; D045744:Cell Line, Tumor; D005260:Female; D019693:Hepatitis, Autoimmune / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D020543:Proteome / Q000032:analysis*
D001323:Autoantibodies; D001324:Autoantigens; D020543:Proteome