CD40 antisense oligonucleotide inhibition of trinitrobenzene sulphonic acid induced rat colitis

• Published in: Gut Vol. 54; no. 1; pp. 70 - 77
• Main Authors: Gao, D, Wagner, A H, Fankhaenel, S, Stojanovic, T, Schweyer, S, Panzner, S, Hecker, M
• Format: Journal Article
• Language: English
• Published: London BMJ Publishing Group Ltd and British Society of Gastroenterology 01-01-2005; BMJ; BMJ Publishing Group LTD; Copyright 2005 by Gut
• Subjects: 6-trinitrobenzene sulphonic acid; Animals; antigen presenting cells; Antigens; antisense oligonucleotide; APC; Biological and medical sciences; CD40; CD40 Antigens - genetics; CD40 Antigens - metabolism; CD40 Ligand - metabolism; Cell adhesion & migration; Cells, Cultured; colitis; Colitis - chemically induced; Colitis - immunology; Colitis - prevention & control; Colon; Crohn Disease - chemically induced; Crohn Disease - immunology; Crohn Disease - prevention & control; Crohns disease; Cytokines; Dendritic cells; Drug Design; EF-2; elongation factor 2; Experiments; Gastroenterology. Liver. Pancreas. Abdomen; Gene Expression Regulation - immunology; IBD; ICAM-1; IFN-γ; IL-12; Immunity, Mucosal; Immunoglobulins; Inflammation; Inflammatory Bowel Disease; Inflammatory diseases; intercellular adhesion molecule 1; interferon γ; interleukin 12; Intestinal Mucosa - immunology; Liposomes; Male; MCP-1; Medical sciences; Molecular weight; monocyte chemoatractive protein 1; Muscle, Smooth, Vascular - immunology; Muscle, Smooth, Vascular - metabolism; ODN; oligonucleotide; Oligonucleotides, Antisense - administration & dosage; Oligonucleotides, Antisense - genetics; Oligonucleotides, Antisense - therapeutic use; Other diseases. Semiology; Pathogenesis; RACE; rapid amplification of cDNA ends; Rats; Rats, Wistar; reverse transcription-polymerase chain reaction; Rodents; RT-PCR; Smooth muscle; Stomach. Duodenum. Small intestine. Colon. Rectum. Anus; Thromboembolism; TNBS; TNF-α; Transfection; trinitrobenzene sulphonic acid; Trinitrobenzenesulfonic Acid; Tumor necrosis factor-TNF; tumour necrosis factor α; vascular cell adhesion molecule; vascular cell adhesion molecule 1; Vascular Cell Adhesion Molecule-1 - metabolism; vascular smooth muscle cells; VCAM-1; VSMC; Germany; Rat; Pathogenesis; Toxicity; Mucosa; Cellular immunity; Inflammatory disease; Prevention; Immunological investigation; Secondary effect; Intestinal disease; Inhibition; Reverse transcription polymerase chain reaction; Humoral immunity; Human; Immune response; Antibody; Interaction; Cytokine; Rodentia; Antisense oligonucleotide; Crohn disease; Anatomic pathology; Vertebrata; Chronic; Mammalia; Treatment; Animal; Digestive diseases; Surgical approach; Colitis; Ulcerative colitis
• ISSN: 0017-5749; 1468-3288; 1458-3288
• DOI: 10.1136/gut.2003.029587

Background: CD154/CD40 interactions play a pivotal role both in humoral and cellular immune responses. Their involvement in the pathogenesis of chronic inflammatory bowel disease (IBD) has been revealed by increased expression of CD40 and CD154 in the inflamed mucosa of patients and the therapeutic effects of anti-CD154 antibodies in experimental colitis. Because of adverse side effects however, the use of such antibodies in patients with IBD may be limited. Aims: An alternative approach to blocking CD154/CD40 interactions by employing a CD40 antisense oligonucleotide (ODN) was explored. Results: After sequencing of the rat CD40 gene, five antisense ODNs were designed, of which one (rAS3) effectively downregulated CD40 expression in rat vascular smooth muscle cells as well as the subsequent changes in gene expression in response to CD40 stimulation. The therapeutic potency of rAS3 was evaluated in the 2,4,6-trinitrobenzene sulphonic acid (TNBS) induced colitis model of the rat. Single intracolonic injection of a liposomal formulation of rAS3 either prior to or post colitis induction markedly suppressed the inflammatory reaction in these animals monitored both macroscopically and microscopically over one week, while application of a scrambled control ODN had no such effects. Moreover, reverse transcription-polymerase chain reaction analyses revealed reduced expression of vascular cell adhesion molecule 1, interleukin 12 p40, and monocyte chemoatractive protein 1 in the inflamed mucosa, which in turn may have contributed to the decrease in leucocyte infiltration judged by immunohistochemistry. Conclusions: These results suggest that CD40 antisense ODNs effectively interfere with CD154/CD40 interactions in vivo and, therefore, may provide a novel approach to the treatment of patients with chronic IBD.