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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
  1. S Sakamakia,
  2. N Takayanagia,
  3. N Yoshizakia,
  4. S Hayashia,
  5. T Takayamaa,
  6. J Katoa,
  7. K Kogawaa,
  8. N Yamauchia,
  9. N Takemotoa,
  10. A Nobuokaa,
  11. T Ayabeb,
  12. Y Kohgob,
  13. Y Niitsua
  1. a4th Department of Internal Medicine, Sapporo Medical University School of Medicine, South-1, West-16, Chuo-ku, Sapporo, 060-8543, Japan, b3rd Department of Internal Medicine, Asahikawa Medical University, Nishikagura 4 sen 5-3-11, Asahikawa, 078-8510, Japan
  1. Dr Y Niitsu. Email: niitsu{at}sapmed.ac.jp

Abstract

BACKGROUND AND AIMS 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 51Cr 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.

  • tropomyosin
  • antibody dependent cell mediated cytotoxicity
  • HLA-DPw9
  • ulcerative colitis
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Footnotes

  • Abbreviations used in this paper:
    TM
    tropomyosin
    UC
    ulcerative colitis
    CD
    Crohn's disease
    PSC
    primary sclerosing cholangitis
    PBC
    primary biliary cirrhosis
    ADCC
    antibody dependent cell mediated cytotoxicity
    AA
    amino acid
    PBS
    phosphate buffered saline
    ALP
    alkaline phosphatase
    PBMNC
    peripheral blood mononuclear cells
    OD
    optical density

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