Expression of a unique protein on colon cancer cells that reacts with a novel monoclonal antibody and ulcerative colitis serum

Abstract

SUMMARY We earlier developed a MoAb, 7E12H12 (IgM isotype), against a protein present in normal colonic epithelial cells. To examine if 7E12H12-reactive protein is expressed in colon cancer cells and is recognized by ulcerative colitis (UC)-associated autoantibody, we investigated several colon cancer cell lines. 7E12H12 reactivity against the cells was examined by indirect immunofluorescence assay and whole cell ELISA against six colon cancer cell lines HT-29, LoVo, COLO 205, DLD-1, LS 180 and SW 1116. A competitive ELISA was developed using 7E12H12 MoAb and patients' serum to examine the cross-reactive antibodies in the serum. Among the six colon cancer cell lines only LS 180, DLD-l and SW 1116 reacted with 7E12H12 MoAb, while others did not. The mean (s.e.m.) inhibition of the binding or 7E12H12 MoAb to LS 180 cells by UC serum (n= 51) was 42±2±1%, whereas in normal subjects (n= 17) it was 14±2±6%, in Crohn's disease (n= 19) it was 15±3±2±5%, in infectious diarrhoea (n= 10) it was 11%±3%, and in systemic lupus erythematosus (n= 10) it was 2%±0±6%. The inhibition by the UC group was significantly (P<0±001 - <0±000l) higher than any of the non-UC groups, and this inhibition was mainly by IgG1 antibody. The protein in the specific colon cancer cells recognized by the 7E12H12 MoAb cross-reacts with UC-IgG1 antibody and may provide an in vitro system to examine the autoimmune mechanisms in UC.