17 th International Conference of the Inflammation Research Association: Wednesday AM (Day 4) September 9-13, 2012
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1 17 th International Conference of the Inflammation Research Association: Wednesday AM (Day 4) September 9-13, 2012 The Sagamore Resort, Bolton Landing, NY, USA The following report includes highlights of Symposium IV entitled Novel Pharmacological Approaches to Inflammatory Bowel Disease. Probiotics and IBD. Karen Madsen of the University of Alberta described the clinical differences between ulcerative colitis (UC) and Cohn s disease (CD). UC presents as continuous superficial inflammation confined to the colon while CD exhibits patchy and transmural inflammation throughout the gut. The incidence of both diseases is increasing. Through gene discovery, key targeted pathways for both diseases have focused on innate (NOD2) and adaptive (IL23R, IL12B, JAK2, STAT3, TNFSF15, CCR6, IL27, and REL) immunity, autophagy (ATG161, IRGM, ATG5), barrier defects (ECM1, CDH1, LAMB, HNF4A, and GNA12), and IL-10 signalling. In general, the causes of IBD appear to be a combination of genetics, environment, immunity, and gut microbial antigens. Diseases influenced by both good and bad gut microbes outside of IBD include autism, asthma, cardiovascular and peripheral vascular disease, colon cancer, biliary disease, and metabolic disease. Types and amounts of microbial species change as one proceeds through the gut. Maintaining a balance between commensals and pathogens is critical to health. In IBD, a severe GI gut dysbiosis (increases in E. Coli, Enterococcus, and Proteobacteria) and pro-inflammatory state exists. This is also seen in the IL-10 deficient mouse model (i.e., the phenotype and severity of colitis noted depends on the type of gut microbiota introduced). These gut bacteria also regulate pathways of T cell differentiation as well as the balance between antiinflammatory and pro-inflammatory cytokine in IBD patients. More specifically, patients with UC and CD exhibit different cytokine profiles, which are TH2 and TH1 driven, respectively. The new paradigm is to try to address the imbalance in gut microflora and the immune dysregulation at the same time and achieve better treatment outcomes. A list of commonly used probiotics was described (i.e., lactobacillus, bifidobacteria, and streptococcus as well as others such as E. Coli Nissle). Probiotics interact with epithelium (as well as the gut lumen, mucus layer, tight junctions, and lamina propria) along the entire GI tract but do not colonize. Effects of probiotics include anti-microbial activity, enhanced barrier integrity, immune response stimulation and immunomodulation, and reduced bacterial adhesion. Effects of probiotics (proor anti-inflammatory) are very much strain dependent.
2 Some IBD patients have altered gut barrier function and increased levels of infiltrating bacterial. The gut barrier is composed of the following barriers designed to keep bacteria out: microbial, chemical, physical, and immunologic. Tight junctions between epithelial cells are extremely important for maintenance of the barrier and are not static. They are affected by ph, oxidative stress, and nutrients as well as microbes and their products. In addition, various receptors, signalling pathways, and cytokines help regulate tight junctions. Increased gut permeability appears to be associated with various diseases including IBD, sepsis, necrotizing pancreatitis celiac disease, type 1 diabetes, food allergy, and alcoholic liver disease. A variety of in vitro and in vivo experiments looking at probiotics and their effects on epithelial resistance and gut permeability were described. Generally, increased gut permeability occurs before the disease and its prevention abolishes the disease. For probiotics to be effective clinically, it is necessary to match the correct probiotic with the appropriate clinical condition and possibly genotype. Melanin Concentrating Hormone (MCH) and Intestinal Inflammation Efi Kokkotou from Harvard described her work in the area of MCH relative to IBD. MHC is a 19 amino acid neuropeptide, that binds to G-protein coupled receptors (MCHR1 and MCHR2) found in brain as well as peripherally. Potential MCH sources in the gut include neuronal and neuroendocrine cells, immune cells, and endothelium. Possible MCH targets in the gut include coloncytes, immune cells (macrophages and T-cells), and fibroblasts. The following were noted: Colonic epithelial and monocytic cells have been shown to express MCHR1 under normal and pro-inflammatory conditions. MHC promotes gut inflammation (primarily by up regulating proinflammatory cytokines while down regulating IL-10) MHC also promotes colonocyte and monocyte survival primarily by inhibiting apoptosis. TH1 and TH2 cells express MCHR1. Likewise, CD4+ T-cell activation up regulates the MCH receptor. MHC promotes fibrosis. Information further suggesting a role for MCH in patients with IBD comes from translational studies where increased expression of MCHR1 in colonic epithelial intestinal cells, myofibroblasts, monocytes, and lamina propria CD4+ cells were noted. Preclinical studies supporting a potential therapeutic role of MHC modulation in IBD were outlined: In MCH knock out mice, TNBS-induced colitis was reduced. An anti-mch antibody reduced colitis in the TNBS model.
3 MCH knock outs exhibit reduced C. difficile toxin-a mediated gut inflammation. Using an anti-mch antibody increased mucosal healing while reducing fibrosis in a chronic colitis model in mice. A small molecule MCH antagonist (DABA-822) reduced experimental colitis. Immune Regulation by the Butyrophilin Family Heather Arnett of Amgen spoke about the role of the butyrophilin family in immune function. She briefly described the importance of regulation of T cells and where the emerging butyrophilin family fits in the scheme. The B7 family (particularly B7-1 and B7-2) appear to play an important costimulatory role in the interaction between APCs and T-cells. The B7 family is part of a superfamily, which includes the butyrophilin family and the butyrophilin-like (BTNL) proteins. The focus of her work has been mostly on the BTNLs and specifically BTNL2. BTNL2 and BTNL9 are all expressed in human B cells while BTNL3 and BTNL8 are expressed in human leukocytes. At the tissue level, BTNL2 is expressed in human small intestine, colon, lymph nodes, spleen, brain, and heart. In the mouse, BTNL2 is expressed in lymphoid tissues (spleen and lymph nodes) and GI tissues (small intestine at the epithelial cell level and Peyer s patch). There appears to be good correlation between what is found in mouse versus the human. A Mdr1a knock out mouse was used to explore the role of BTNL2 in IBD. This animal lacks the p-glycoprotein transporter and spontaneously develops colitis within 4-6 months of age. It has been demonstrated that BTNL2 expression is up regulated in Mdr1a knock out mice. Using mubtnl2-fc protein as a tool, dose-dependent suppression of murine CD4+ splenocytes, mesenteric lymph node, and Peyer s patch proliferation was noted in the absence of effects on B cell proliferation. Mouse BTNL2-Fc also inhibited human CD4+ T cell proliferation. Some other points made included: BTNL2 inhibits markers of T cell activation including IL-2, PD-1, NFATc1, and IFN-. mubtnl2-fc inhibits cytokine production under in vitro conditions including TNF, GM-CSF, IL-10, IL-4, IL-2, IFN, IL-6 and IL-17. BTNL2 induces Foxp3+ CD4 T cells expressing Treg surface markers.
4 The Role of IL-22 in Mucosal Immunity The final speaker, Wenjun Ouyang from Genentech, addressed the pathogenic role of IL-22 in various inflammatory diseases processes. IL-22 is part of the IL-10 family of cytokines, which includes IL-10, IL-19, IL-20, IL-24, IL-26, IL-28, and IL-29. IL-22 is produced by various leukocytes including TH17, TH22, NK, LTi, and DCs. IL-22R is expressed in various tissue epithelial cells including keratinocytes, intestinal epithelial cells, lung epithelial cells, hepatocytes, and acinar cells. Binding and stimulation of the IL-22R results in downstream Stat3 activation and may stimulate other downstream effectors including Stat1, Stat5, and MEK. Differentiation of CD4 T cells to various T helper cells (Th1, Th17, Th2, TFH, and Treg) produces a number of cytokines that effect the following either positively or negatively: 1) cellular immunity (IFN, IL-2, LT), 2) tissue repair and bacterial infection (IL-17, IL-21, IL- 22), 3) humoral immunity and parasitic infection (IL-4, IL-5, IL-13), 4) antibody production (IL-21), and 5) immune repression (IL-10, TGF). Under in vitro conditions, IL-23 has been shown to stimulate IL-22 production in various leukocytes populations (T cells, CD8 T cells, monocytes, and CD4 T cells) as well as innate lymphoid cells. The question remains as to whether IL-22 has direct antiinflammatory /tissue protective effect. Work in the area of psoriasis was summarized. It appears that IL-22 and IL-22R are highly expressed in primary human psoriatic keratinocytes. Using an epidermal organ culture system, IL-22 produced many of the characteristics associated with psoriatic skin including keratinocyte hyperplasia. The pathogenic role of IL-22 in psoriasis was discussed. Through IL-23-induced activation of CD8+, Th1, TH17, and TH22, IL-22 was produced. IL-22 induced a pathologic response characterized by parakeratosis, hypogranulosis, production of antimicrobial peptides, acanthosis, chemokine production, and wound healing. Switching back to IL-22 and the gastrointestinal tract, IL-22R is expressed on intestinal epithelial cells in both mouse and human. Using an attaching and effacing (A/E) bacteria infection model (C. rodentium) or dextran sulphate (DSS) model in mice, the following observations (not all inclusive) were made: A/E Model Infection with A/E bacterial pathogen induces IL-22. IL-23 is necessary for IL-22 induction. IL-22 is necessary for host against A/E bacterial infection. Downstream targets of IL-22 include RegIII and RegIII IL-22 is only required during the initial infection phase. Neither T nor B cells are necessary for IL-22 mediated immunity. IL-22 is downstream of the LT pathway during infection.
5 LT is required for induction of IL-22 and maintenance of lymphoid follicles in the colon during infection. IL-22 is necessary for maintenance of lymphoid follicles during infection DSS Model IL-22/Stat3 plays a primary role in epithelial healing and preservation of epithelial integrity. Exogenously administered IL-22 can reduce disease severity. IL-22 preserves goblet cell function.
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