Treatment of murine Th1- and Th2-mediated inflammatory bowel disease with NF- B decoy oligonucleotides

Abstract

The Th1 and Th2 T cell responses that underlie inflammatory bowel diseases (IBDs) are likely to depend on NF-κB transcriptional activity. We explored this possibility in studies in which we determined the capacity of NF-κB decoy oligodeoxynucleotides (decoy ODNs) to treat various murine models of IBD. In initial studies, we showed that i.r. (intrarectal) or i.p. administration of decoy ODNs encapsulated in a viral envelope prevented and treated a model of acute trinitrobenzene sulfonic acid-induced (TNBS-induced) colitis, as assessed by clinical course and effect on Th1 cytokine production. In further studies, we showed that NF-κB decoy ODNs were also an effective treatment of a model of chronic TNBS-colitis, inhibiting both the production of IL-23/IL-17 and the development of fibrosis that characterizes this model. Treatment of TNBS-induced inflammation by i.r. administration of NF-κB decoy ODNs did not inhibit NF-κB in extraintestinal organs and resulted in CD4 + T cell apoptosis, suggesting that such treatment is highly focused and durable. Finally, we showed that NF-κB decoy ODNs also prevented and treated oxazolone-colitis and thus affect a Th2-mediated inflammatory process. In each case, decoy administration led to inflammation-clearing effects, suggesting a therapeutic potency applicable to human IBD.

NOD2 Transgenic Mice Exhibit Enhanced MDP-Mediated Down-Regulation of TLR2 Responses and Resistance to Colitis Induction

“…TNBS-colits was induced by intrarectal administration of 3.75 mg of TNBS (Sigma) dissolved in 45% ethanol as previously described (16). Challenged mice were observed for 4-5 days and then sacrificed for histopathologic assessment and harvest of colonic LPMCs as previously described (16).…”

“…TNBS-colits was induced by intrarectal administration of 3.75 mg of TNBS (Sigma) dissolved in 45% ethanol as previously described (16). Challenged mice were observed for 4-5 days and then sacrificed for histopathologic assessment and harvest of colonic LPMCs as previously described (16). LPMCs were cultured in 48-well plates (10 6 cells/ml) in wells coated with anti-CD3ε (10μg/ ml, Pharmingen) and soluble anti-CD28 (1μg/ml, Pharmingen) for IFN-γ production, and with IFNγ (1000 units/ml, Pharmingen) and Staphylococcus aureus (1:10,000 Calbiochem) for IL-12p70 production.…”

“…PCR with these primers introduced L980P followed by stop codon TAA in the mouse NOD2 cDNA that was equivalent to the human 3020insC Crohn disease frame-shift mutation (1,2). For in vivo delivery of these plasmids, large amounts of plasmid was prepared using an endofree Plasmid Mega kit (Qiagen), and then packaged into a Hemagglutinin Virus Japan-Envelope vector using protamine sulfate (HVJ-E, GenomIdea, Osaka, Japan) according to the manufacturer's instructions (16). Mice induced with TNBS-colitis were subjected to IP administration of 100 μg of plasmid DNA/mouse encapsulated in 5000 HAU of HVJ-E vector in 100μl of PBS at −1, 0 and +1 day of TNBS intrarectal challenge.…”

“…The second model of inflammation studied was trinitrobenzene sulfonic acid (TNBS)-colitis, a Th1 colitis that is initiated by TNP-substituted colonic proteins (11,16). In studies of TNBScolitis we administered intra-rectal TNBS (3.75 mg) in 45% ethanol or ethanol alone to C57BL/ 10 NOD2-Tg mice and littermate control mice and then characterized the colitis in these mouse groups.…”

Insights from advances in research of chemically induced experimental models of human inflammatory bowel disease

“…Nuclear factor (NF)-κB is the key transcription factor for pro-inflammatory responses, and is thought to be important in the initiation and progression of both human IBD and animal models of colitis [65,66] . Disease activity in mice with TNBS-induced colitis is inhibited by antisense oligonucleotides that inhibit the p65 subunit of NF-κB, which suggests a critical role for NF-κB in mediating inflammatory responses [65] .…”