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.
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.
Hibi
T
T
Department of Internal Medicine, School of Medicine, Keio University, Japan.
Ohara
M
M
Kobayashi
K
K
Brown
W R
WR
Toda
K
K
Takaishi
H
H
Hosoda
Y
Y
Hayashi
A
A
Iwao
Y
Y
Watanabe
M
M
D000293:Adolescent; D000328:Adult; D000906:Antibodies / Q000032:analysis*; D001323:Autoantibodies / Q000032:analysis; D002460:Cell Line; D003093:Colitis, Ulcerative / Q000276:immunology*; D003106:Colon / Q000276:immunology*; D003109:Colonic Diseases, Functional / Q000276:immunology*; D003424:Crohn Disease / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D008875:Middle Aged; D009077:Mucins / Q000276:immunology*; D011233:Precipitin Tests
D000906:Antibodies; D001323:Autoantibodies; D009077:Mucins