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. 1999 Feb;44(2):196–202. doi: 10.1136/gut.44.2.196

Detection of anti-colon antibodies in inflammatory bowel disease using human cultured colonic cells

J Lee 1, A Cevallos 1, A Naeem 1, J Lennard-Jones 1, M Farthing 1
PMCID: PMC1727366  PMID: 9895378

Abstract

Background—Investigation of anti-colon antibodies may be simplified if a sensitive method and homogeneous source of antigen were available. 
Aims—To examine the anti-colon antibody response using human colonic carcinoma cell lines as antigen. 
Subjects—Patients with inflammatory bowel disease and other gastrointestinal disorders and healthy controls were studied. 
Methods—Comparative enzyme linked immunosorbent assays (ELISAs) were performed to assess the value of whole Caco-2, HT-29, and LS-180 cells as antigen. The antigenic determinants of the immune response were characterised by western blot analysis. 
Results—Sera demonstrated immunoreactivity against each of the cell lines, but different epitopes were recognised. Applying whole Caco-2 cells as antigen in an ELISA, the prevalence of anti-colon antibodies was significantly greater in patients with ulcerative colitis (36%) than Crohn's disease (13%), other gastrointestinal disorders (13%) and healthy controls (0) (p<0.05). The immune response was not associated with one predominant antigen. 
Conclusions—Fixed whole cell ELISA is a simple and feasible method for studying the anti-colon antibody response. This response is non-specific, being directed against multiple antigens, and is likely to be an epiphenomenon of inflammatory bowel disease, more so for ulcerative colitis than Crohn's disease. 



Keywords: anti-colon antibodies; inflammatory bowel disease

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Figure 1 .

Figure 1

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Protein profile of LS-180, HT-29 and Caco-2 cell homogenates separated by 5% stacking-10% separating polyacrylamide gels and stained with Coomassie blue. Bands corresponding to similar molecular masses are seen for each of the cells. 


Figure 2 .

Figure 2

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Examples of western blot analysis for sera from three patients with ulcerative colitis, (A) (B) and (C). Although the protein profiles of the cell lines are similar (see fig 1) and the nitrocellulose strips were incubated with the same serum sample, circulating antibodies did not recognise protein bands associated with all cells. 


Figure 3 .

Figure 3

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Scattergram of absorbance values obtained by ELISA using fixed whole Caco-2 cells as antigen to investigate the prevalence of anti-colon antibodies in patients with inflammatory bowel disease (IBD). Values above the dotted line are considered positive for anti-colon antibodies, representing results greater than three standard deviations above the mean of the healthy control group. CD, Crohn's disease; UC, ulcerative colitis. 


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