Autoantibodies against the specific epitope of human tropomyosin(s) detected by a peptide based enzyme immunoassay in sera of patients with ulcerative colitis show antibody dependent cell mediated cytotoxicity against HLA-DPw9 transfected L cells.
OBJECTIVE
Recent studies suggest that tropomyosin (TM) may act as a putative autoantigen in ulcerative colitis (UC). Recently, we identified, by computer homology analysis, a specific peptide (HIAEDADRK) in human TM that can bind to HLA-DPw9. The aim of this study was to investigate the presence of autoantibodies against this peptide in UC.


METHODS
Antibodies were measured by ELISA with a synthetic peptide in 20 healthy volunteers, 48 patients with UC, 26 with Crohn's disease (CD), eight with primary sclerosing cholangitis (PSC), and six with primary biliary cirrhosis (PBC). The functional significance of antibodies was investigated by antibody dependent cell mediated cytotoxicity (ADCC) against DPw9 transfected L cells using a standard (51)Cr release assay.


RESULTS
Optical density values (mean (SD)) of sera from patients with UC (1.40 (0. 52)) and PSC (1.65 (0.12)) were significantly higher than those from healthy volunteers (0.32 (0.28)) (p<0.05), CD (0.50 (0.34)) (p<0.05) and PBC (0.14 (0.09)) (p<0.05). Values in UC decreased with clinical improvement. The ADCC activity of UC sera correlated well with antibody titre against this synthetic peptide.


CONCLUSIONS
Anti-TM antibody was detected in UC sera by a specific peptide based ELISA with high reproducibility. This peptide may be an antigenic epitope of TM involved in the immunopathogenesis of UC and, perhaps, PSC.
Sakamaki
S
S
4th Department of Internal Medicine, Sapporo Medical University School of Medicine, Japan.
Takayanagi
N
N
Yoshizaki
N
N
Hayashi
S
S
Takayama
T
T
Kato
J
J
Kogawa
K
K
Yamauchi
N
N
Takemoto
N
N
Nobuoka
A
A
Ayabe
T
T
Kohgo
Y
Y
Niitsu
Y
Y
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology*; D016022:Case-Control Studies; D015209:Cholangitis, Sclerosing / Q000276:immunology; D003093:Colitis, Ulcerative / Q000276:immunology*; D003424:Crohn Disease / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000276:immunology; D005260:Female; D006682:HLA-DP Antigens / Q000276:immunology*; D006801:Humans; D007739:L Cells / Q000276:immunology; D008105:Liver Cirrhosis, Biliary / Q000276:immunology; D051379:Mice; D008875:Middle Aged; D015203:Reproducibility of Results; D014335:Tropomyosin / Q000276:immunology*
D001323:Autoantibodies; D000939:Epitopes; D006682:HLA-DP Antigens; D014335:Tropomyosin