Isolation and characterization of a novel short peptide associated with Crohn's disease

• Published in: Clinical and experimental immunology Vol. 166; no. 1; pp. 72 - 79
• Main Authors: Mitsuyama, K., Niwa, M., Masuda, J., Kuwaki, K., Yamasaki, H., Takedatsu, H., Kobayashi, T., Sata, M.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.10.2011; Blackwell; Oxford University Press; Blackwell Science Inc
• Subjects: Adult; Analytical, structural and metabolic biochemistry; Antibodies - blood; Antibodies - immunology; Biological and medical sciences; biomarker; Caco-2 Cells; Case-Control Studies; Colitis, Ulcerative - blood; Colitis, Ulcerative - diagnosis; Colitis, Ulcerative - immunology; Colitis, Ulcerative - pathology; Colonic Neoplasms - blood; Colonic Neoplasms - diagnosis; Colonic Neoplasms - immunology; Colonic Neoplasms - pathology; Crohn Disease - blood; Crohn Disease - diagnosis; Crohn Disease - immunology; Crohn Disease - pathology; Crohn's disease; DNA, Complementary - analysis; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Fundamental and applied biological sciences. Psychology; Gastroenterology. Liver. Pancreas. Abdomen; Humans; Interleukin-1beta - biosynthesis; Interleukin-1beta - secretion; Interleukin-6 - biosynthesis; Interleukin-6 - secretion; Leukocytes, Mononuclear - immunology; Leukocytes, Mononuclear - pathology; Male; Medical sciences; Other diseases. Semiology; Peptide Library; Peptides - chemistry; Peptides - immunology; Peptides - isolation & purification; Peptides - pharmacology; phage display; ROC Curve; Sequence Analysis, DNA; Serum - chemistry; Stomach. Duodenum. Small intestine. Colon. Rectum. Anus; Translational Studies; Tumor Necrosis Factor-alpha - biosynthesis; Tumor Necrosis Factor-alpha - secretion; Peptides; Biological marker; Biochemistry; Crohn's disease; biomarker; Inflammatory disease; Characterization; Crohn disease; Phage display; Association; Digestive diseases; Intestinal disease; Isolation
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1111/j.1365-2249.2011.04444.x

Summary Phage display technology has been utilized to select target molecules against circulating antibodies. The aims of this study were to isolate a peptide that binds with serum from Crohn's disease (CD) patients and to examine its diagnostic and pathogenic significance. A phage display library was constructed using cDNA from Caco‐2 cells. Affinity selection using this cDNA library and serum samples from patients with CD was then performed. Phage clones that specifically reacted with the CD sera were then selected using a phage enzyme‐linked immunosorbent assay (ELISA). After the DNA sequences of the selected phages were determined and converted to amino acid sequences, the synthesized peptides were examined using an ELISA. The effect of the synthesized peptides on cytokine release from cultured blood mononuclear cells was investigated. An ELISA analysis for TCP‐353 demonstrated that while 61·7% of the samples from CD patients were seroreactive, seroreactivity was less common among patients with ulcerative colitis (7·3%), acute colitis (0%) or colon cancer (11·4%) and among normal subjects (2·8%). The induction of interleukin (IL)‐1β, IL‐6 and tumour necrosis factor (TNF)‐α release, but not IL‐10 release, in response to TCP‐353 peptide was enhanced in CD mononuclear cells only. We isolated a novel peptide that specifically binds to CD sera and stimulates the proinflammatory responses of CD mononuclear cells. TCP‐353 may have diagnostic, pathogenic and therapeutic significance with regard to the treatment of CD.