Immunoproteomic to Identify Antigens in the Intestinal Mucosa of Crohn's Disease Patients

Abstract

Incidences of Crohn disease (CD) have increased significantly in the last decade. Immunoproteomics are a promising method to identify biomarkers of different diseases. In the present study, we used immunoproteomics to study proteins of intestinal mucosal lesions and neighboring normal intestinal mucosa of 8 CD patients. Reactive proteins were validated by Western blotting. Approximately 50 protein spots localized in the 4 to 7 pI range were detected on two-dimensional electrophoresis gels, and 6 differentially expressed protein spots between 10 and 100 kDa were identified. Reactive proteins were identified as prohibitin, calreticulin, apolipoprotein A-I, intelectin-1, protein disulfide isomerase, and glutathione s-transferase Pi. Western blotting was conducted on the intestinal mucosa of another 4 CD patients to validate the reactive proteins. We found that intestinal mucosal lesions had high levels of prohibitin expression. Glutathione s-transferase expression was detected in 100% of the intestinal mucosa examined. Thus, we report 6 autoantigens of CD, including 3 new and 3 previously reported autoantigens. Intelectin-1, protein disulfide isomerase, and glutathione-s-transferases may be used as biomarkers for CD pathogenesis.

Use of immunoproteomics to identify immunogenic proteins in a rat model of acute respiratory distress syndrome

“…Protein separation by two-dimensional polyacrylamide gel electrophoresis . Lung tissue proteins from each group (n=6/group) were pooled and separated by 2-D PAGE as previously described (14,19). A total of 100 µg of protein was mixed with rehydration buffer [8 M urea, 2% CHAPS, 20 mM DTT, 0.5% IPG buffer (pH 3-10) and 0.001% trace bromophenol blue] and applied to IPG strips (pH 3-10; 13 cm; GE Healthcare Life Sciences, Uppsala, Sweden).…”

“…Immunoproteomics is a technique that involves the separation of proteins by two-dimensional electrophoresis (2-DE) followed by western blotting, and has been used previously to identify immunogenic proteins in various diseases (14,15).…”

A comparative review of intelectins

“…In normal human tissues, hITLN1 is mainly secreted by goblet cells in the gastrointestinal tract into the intestinal lumen. Researchers have discovered that hITLN1 was overexpressed in most gastric cancer tissues, and the overexpression level was associated with the classification of gastric adenocarcinoma and was negatively correlated with lymph node metastasis, distant metastasis and tumour infiltration 99 . hITLN1 inhibited the nuclear transport and transcriptional activity of β‐catenin in gastric cancer cells by upregulating hepatocyte nuclear factor 4 alpha (HNF4α).…”

Mass spectrometry-based proteomic exploration of the human immune system: focus on the inflammasome, global protein secretion, and T cells

“…Proteomics has also been used to identify disease mechanisms and novel biomarkers and from mucosal colon biopsies for inflammatory bowel disease (IBD) including Crohn's disease (CD) and ulcerative colitis (UC) [69][70][71][72][73] …”