Immunostimulatory oligonucleotides inhibit colonic proinflammatory cytokine production in ulcerative colitis

Background: We previously showed that Toll-like receptor-9 (TLR-9) ligands ameliorate experimental colitis. In this study, we evaluated the effect of TLR-9 ligands on the generation of proinflammatory cytokines by human colonic mucosa. Materials and Methods: Colonoscopic biopsies were obtained from patients with active ulcerative colitis (UC) and from normal subjects. The tissue was organ cultured for 24 hours in the presence or absence of different types of immunostimulatory (ISS) (CpG)oligonucleotides (ODNs). Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) levels in the medium were determined by enzyme-linked immunosorbent assay. Results: in active UC, hTNF-alpha and hIL-1 beta generation by inflamed colonic mucosa is 7- and 3-fold higher, respectively, than their generation by normal mucosa. Class B CpG ODNs inhibited colonic TNF-alpha and IL-1 beta generation by 50%, whereas class A or C ODNs had a partial or no effect, respectively. A novel class of ODNs that is based on multiple TCG repeats was as effective as class B ODNs. This inhibition resulted from the transcriptional suppression of IL-1 beta that occurred within the first 2 hours after ISS-ODN incubation. The addition of chloroquine abolished the inhibitory effects of ISS-ODNs on colonic TNF-alpha and IL-1 beta generation. Conclusions: Only certain classes of ISS-ODNs inhibit the enhanced TNF-alpha and IL-1 beta generated ex vivo by inflamed colonic mucosa of patients with UC. The effect of ISS-ODNs is mediated by triggering of TLR-9. These results suggest a potential therapeutic value for ISS-ODN's in UC.