Skip to main content
Gut logoLink to Gut
. 1994 Feb;35(2):224–230. doi: 10.1136/gut.35.2.224

Enzyme linked immunosorbent assay (ELISA) and immunoprecipitation studies on anti-goblet cell antibody using a mucin producing cell line in patients with inflammatory bowel disease.

T Hibi 1, M Ohara 1, K Kobayashi 1, W R Brown 1, K Toda 1, H Takaishi 1, Y Hosoda 1, A Hayashi 1, Y Iwao 1, M Watanabe 1, et al.
PMCID: PMC1374498  PMID: 8307474

Abstract

Circulating anti-goblet cell antibody and its corresponding antigen in patients with inflammatory bowel disease were investigated. Anti-goblet cell antibody in the serum was examined by immunocytochemistry and enzyme linked immunosorbent assay (ELISA), using a colonic cancer cell line, HT29-18-N2, which differentiates into intestinal goblet cells. The frequencies of anti-goblet cell antibody detected by immunocytochemistry were 14 in 48 patients with ulcerative colitis (29%) and five in 15 patients with Crohn's disease (33%). By ELISA, the frequencies of anti-goblet cell antibody were 38% in ulcerative colitis and 33% in Crohn's disease. This antibody did not relate directly to anti-neutrophil cytoplasmic antibodies (ANCA), although the serum samples positive for anti-goblet cell antibody were commonly positive for ANCA in ulcerative colitis. Immunoprecipitation and SDS polyacrylamide gel electrophoresis (PAGE) study showed that the antibody in the ELISA positive serum samples recognised a > 200 kD goblet cell antigen, which remained unchanged after reduction, indicating that it consists of single chain polypeptides. These results suggest that there is a subgroup of inflammatory bowel disease that has circulating anti-goblet cell antibody reactive with a > 200 kD antigen. The antibody detected by newly established ELISA will be a disease marker for this group and the identification of the corresponding antigens may be important for the understanding of the underlying immune abnormalities.

Full text

PDF
228

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Asakura H., Tsuchiya M., Aiso S., Watanabe M., Kobayashi K., Hibi T., Ando K., Takata H., Sekiguchi S. Association of the human lymphocyte-DR2 antigen with Japanese ulcerative colitis. Gastroenterology. 1982 Mar;82(3):413–418. [PubMed] [Google Scholar]
  2. BROBERGER O., PERLMANN P. Autoantibodies in human ulcerative colitis. J Exp Med. 1959 Nov 1;110:657–674. doi: 10.1084/jem.110.5.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BROBERGER O., PERLMANN P. In vitro studies of ulcerative colitis. I. Reactions of patients' serum with human fetal colon cells in tissue cultures. J Exp Med. 1963 May 1;117:705–716. doi: 10.1084/jem.117.5.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cambridge G., Rampton D. S., Stevens T. R., McCarthy D. A., Kamm M., Leaker B. Anti-neutrophil antibodies in inflammatory bowel disease: prevalence and diagnostic role. Gut. 1992 May;33(5):668–674. doi: 10.1136/gut.33.5.668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chapman R. W., Cottone M., Selby W. S., Shepherd H. A., Sherlock S., Jewell D. P. Serum autoantibodies, ulcerative colitis and primary sclerosing cholangitis. Gut. 1986 Jan;27(1):86–91. doi: 10.1136/gut.27.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Das K. M., Dubin R., Nagai T. Isolation and characterization of colonic tissue-bound antibodies from patients with idiopathic ulcerative colitis. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4528–4532. doi: 10.1073/pnas.75.9.4528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Das K. M., Sakamaki S., Vecchi M., Diamond B. The production and characterization of monoclonal antibodies to a human colonic antigen associated with ulcerative colitis: cellular localization of the antigen by using the monoclonal antibody. J Immunol. 1987 Jul 1;139(1):77–84. [PubMed] [Google Scholar]
  8. Duerr R. H., Targan S. R., Landers C. J., Sutherland L. R., Shanahan F. Anti-neutrophil cytoplasmic antibodies in ulcerative colitis. Comparison with other colitides/diarrheal illnesses. Gastroenterology. 1991 Jun;100(6):1590–1596. doi: 10.1016/0016-5085(91)90657-7. [DOI] [PubMed] [Google Scholar]
  9. Hibi T., Aiso S., Ishikawa M., Watanabe M., Yoshida T., Kobayashi K., Asakura H., Tsuru S., Tsuchiya M. Circulating antibodies to the surface antigens on colon epithelial cells in ulcerative colitis. Clin Exp Immunol. 1983 Oct;54(1):163–168. [PMC free article] [PubMed] [Google Scholar]
  10. Hibi T., Aiso S., Yoshida T., Watanabe M., Asakura H., Tsuru S., Tsuchiya M. Anti-colon antibody and lymphocytophilic antibody in ulcerative colitis. Clin Exp Immunol. 1982 Jul;49(1):75–80. [PMC free article] [PubMed] [Google Scholar]
  11. Hibi T., Ohara M., Toda K., Hara A., Ogata H., Iwao Y., Watanabe N., Watanabe M., Hamada Y., Kobayashi K. In vitro anticolon antibody production by mucosal or peripheral blood lymphocytes from patients with ulcerative colitis. Gut. 1990 Dec;31(12):1371–1376. doi: 10.1136/gut.31.12.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Huet C., Sahuquillo-Merino C., Coudrier E., Louvard D. Absorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation. J Cell Biol. 1987 Jul;105(1):345–357. doi: 10.1083/jcb.105.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kobayashi K., Blaser M. J., Brown W. R. Identification of a unique IgG Fc binding site in human intestinal epithelium. J Immunol. 1989 Oct 15;143(8):2567–2574. [PubMed] [Google Scholar]
  14. Lagercrantz R., Hammarström S., Perlmann P., Gustafsson B. E. Immunological studies in ulcerative colitis. 3. Incidence of antibodies to colon-antigen in ulcerative colitis and other gastro-intestinal diseases. Clin Exp Immunol. 1966 Jul;1(3):263–276. [PMC free article] [PubMed] [Google Scholar]
  15. Marcussen H. Fluorescent anti-colonic and E. coli antibodies in ulcerative colitis. Scand J Gastroenterol. 1978;13(3):277–281. doi: 10.3109/00365527809179820. [DOI] [PubMed] [Google Scholar]
  16. Marcussen H., Nerup J. Fluorescent anti-colon and organ-specific antibodies in ulcerative colitis. Scand J Gastroenterol. 1973;8(1):9–15. [PubMed] [Google Scholar]
  17. Nakane P. K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. doi: 10.1177/22.12.1084. [DOI] [PubMed] [Google Scholar]
  18. Roche J. K., Fiocchi C., Youngman K. Sensitization to epithelial antigens in chronic mucosal inflammatory disease. Characterization of human intestinal mucosa-derived mononuclear cells reactive with purified epithelial cell-associated components in vitro. J Clin Invest. 1985 Feb;75(2):522–530. doi: 10.1172/JCI111728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Saxon A., Shanahan F., Landers C., Ganz T., Targan S. A distinct subset of antineutrophil cytoplasmic antibodies is associated with inflammatory bowel disease. J Allergy Clin Immunol. 1990 Aug;86(2):202–210. doi: 10.1016/s0091-6749(05)80067-3. [DOI] [PubMed] [Google Scholar]
  20. Shanahan F., Duerr R. H., Rotter J. I., Yang H., Sutherland L. R., McElree C., Landers C. J., Targan S. R. Neutrophil autoantibodies in ulcerative colitis: familial aggregation and genetic heterogeneity. Gastroenterology. 1992 Aug;103(2):456–461. doi: 10.1016/0016-5085(92)90834-l. [DOI] [PubMed] [Google Scholar]
  21. Takahashi F., Das K. M. Isolation and characterization of a colonic autoantigen specifically recognized by colon tissue-bound immunoglobulin G from idiopathic ulcerative colitis. J Clin Invest. 1985 Jul;76(1):311–318. doi: 10.1172/JCI111963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Takahasi F., Shah H. S., Wise L. S., Das K. M. Circulating antibodies against human colonic extract enriched with a 40 kDa protein in patients with ulcerative colitis. Gut. 1990 Sep;31(9):1016–1020. doi: 10.1136/gut.31.9.1016. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Toyoda H., Wang S. J., Yang H. Y., Redford A., Magalong D., Tyan D., McElree C. K., Pressman S. R., Shanahan F., Targan S. R. Distinct associations of HLA class II genes with inflammatory bowel disease. Gastroenterology. 1993 Mar;104(3):741–748. doi: 10.1016/0016-5085(93)91009-7. [DOI] [PubMed] [Google Scholar]
  24. Vecchi M., Sakamaki S., Diamond B., Novikoff A. B., Novikoff P. M., Das K. M. Development of a monoclonal antibody specifically reactive to gastrointestinal goblet cells. Proc Natl Acad Sci U S A. 1987 May;84(10):3425–3429. doi: 10.1073/pnas.84.10.3425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wright R., Truelove S. C. Auto-immune reactions in ulcerative colitis. Gut. 1966 Feb;7(1):32–40. doi: 10.1136/gut.7.1.32. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Gut are provided here courtesy of BMJ Publishing Group

RESOURCES