Disclosures. Spondyloarthropathy. Spondyloarthropathy SpA shared distinctive clinical features. Spondyloarthropathy 11/13/2012

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1 // Protective and pathogenic effects of IL- related cytokines in the SKG mouse model of and Ileitis Nil Disclosures Helen Benham, Linda Rehaume, Merja Ruutu, Jared Velasco, Kristine Kikly, Geoffrey Strutton, Sumaira Hasnain, Michael McGuckin, Matthew Brown and Ranjeny Thomas SpA are common -% of population affected Progressive chronic conditio Prognosis variable Treatment optio suboptimal SpA coists of several rheumatic conditio Ankylosing Spondylitis Psoriatic Arthritis Reactive Arthritis Arthropathy of inflammatory bowel disease Undifferentiated Juvenile SpA shared distinctive clinical features Grouped together on the basis of shared distinctive clinical features and genetic susceptibility Reveille, J. D. Nat. Rev. Rheumatol. Uptodate.com.au Uptodate.com.au SpA shared genetic susceptibility HLA largest component of genetic susceptibility in SpA Class I MHC (HLA-C) - main role in psoriasis HLA-B - AS-like disease Psoriasis, AS and Crohn s disease all associated IL-, IL-B, IL-R, Card9, PTGER- JAK and STAT Strongly implicates the Th/IL- axis pathogenesis of SpA and related skin, eye and gut diseases IL- Axis in SpA Th and Innate cells both gut and joint Th -IL-+ IL-- IFNγ- -CCR+CCR+CD+ -IL-R+ -Skin/Joint homing Lttman DR & Rudeky AY Cell Peck A et.al Clin Immunol 9 Burton PR et.al Nat. Genet. & Reveille JD et.al Nat. Genet. & Reveille JD Nat.Rev.Rheumatol. Ouyang W et.al Immunity 8 Lories RJ & McInnes IB Nat.Med. & Sherlock JP et.al Nat.Med.

2 Ileum IL- per mg of tissue IL- mrna expression /HPRT // SKG model of SpA Zap- mutation Dectin- is a receptor for β-glucan Auto-reactive T cells Susceptibility genes are dowtream of the dectin- Susceptibility to autoimmunity -syk pathway Sakaguchi APCs promote expression of IL-β, IL-p, IL- spontaneous RA like disease in setting p, IL-p9 of fungal lung infection Signalling through IL-R activates Jak and STAT IL- is required for expaion of Th cells and Thomas innate cell activation fungal-β-glucan induced SpA like disease SKG model of SpA Clinical and histological phenotype closely resembles human SpA Ruutu M et.al Arthritis and Rheumatism Sakaguchi et al.febs Letters Reid DM et.al Curr Opin Immunol 9 Axial and peripheral joint inflammation in the SKG mouse model of SpA is IL- dependent Treatment with Anti-IL Ab after the initiation of disease improves axial and peripheral joint disease Ankle Joint Isotype Anti-ILAb Isotype Anti-ILAb Enthesis Spine Isotype Anti-ILAb SKG Tx curdlan SKG + Anti-ILAb Isotype Anti-ILAb Ileitis severity is decreased with Anti-IL Ab given from initiation of disease and in a treatment regime IL- mrna and protein are expressed within days of induction and increased over time in the ileum but not in serum, lymph nodes or joint SKG ileum + curdlan Balb/c ileum + curdlan SKG and Balb/c joint and serum + curdlan..8 Isotype Anti-IL Ab... Day Day Day Day. d d d d d d d d d d d d d d d Ileum Iliac LN Mes LN Spleen Liver SKG + Curdlan SKG + Anti-ILAb

3 % of initial body weight Clinical score - paw width (mm) 8 Average Paw Width (mm) // IL-A-/- SKG mice have decreased joint severity compared to SKG wt treated with curdlan IL-A-/- SKG mice are protected from severe enthesitis in peripheral joints. IL-A -/- + curdlan Peripheral enthesitis histology Weeks after curdlan IL-A -/- IL-A-/- SKG wt peripheral enthesis IL-A-/- SKG peripheral enthesis SKG IL-A -/ IL SKG CD+ T cell trafer WT SKG CD+ T cell trafer 8 9 Weeks after trafer Peripheral enthesitis histology SKG CD+ IL-A -/- CD+ IL-A-/- SKG mice have decreased severity of Ileitis and treatment with Anti-IL-Ab may reduce severe disease SKG mice treated with Anti-ILAb have modestly suppressed arthritis and protected entheses IL-A-/- Isotype Anti-ILAb Peripheral Enthesitis Histology Isotype Anti-IL Ab Isotype Anti-IL Ab Anti-IL Ab Neutralising IL- exacerbates ileitis in SKG mice IL- appears to play a protective role in the early development of gut disease Isotype IL- SKG + Anti-ILAb 9 SKG + Curdlan 8 Weeks after curdlan Isotype Anti-IL Ab SKG +Anti-IL Ab

4 mrna expression ileum mrna expression ileum mrna expression ileum Goblet cell/ cypt villus units // Neutralising IL- in the SKG IL-A-/- mice had no effect on the development of joint disease or ileitis Protective and pathogenic effects of the IL- related cytokines in the SKG mouse model of Fungal-B glucan SKG ZAP- mutation Auto reactive T cells Signalling through Dectin- Inflammatory cascade involving IL- /Th axis SpA Axial and peripheral IBD Arthritis and Ileitis are IL- dependent Histology Ileum IL- is required for severe disease of joints and ileum with a particular role in the peripheral entheses IL- also likely induces disease at the peripheral entheses and plays a protective role in the gut IL- is likely from the gut IL-A -/- IL-A -/- + Anti-ILAb IL-A -/- + Isotype IL-A-/- IL-A-/- + Anti-ILAb IL-A -/- Isotype IL-A-/- IL-A-/- + Anti-IL Ab IL--/- + Isotype ER Stress Proinflammatory cytokine amplification Mucosal barrier functio ER stress in the ileum of SKG mice is associated to altered IL- production Grp8 expression Ileum Day Day Day Day sxbp expression Ileum Day Day Day Day Balb/c SKG + Anti IL- Ab 8 IL- alters pro-inflammatory cytokine production from the ileum and systemically at initiation of disease in SKG mice IL- Ileum D D D D D D D IL- serum SKG + Anti-ILAb TNFa ileum D D D D D D D TNFa serum SKG + Anti-ILAb IFNg ileum D D D D D D D IFNg serum SKG + Anti-ILAb D D D D D D D D D D D D D D D D D D D D D Sumaira Hasnain SKG + Anti-ILAb SKG + Anti-ILAb SKG + Anti-ILAb Anti-IL- increases muc- expression and correlates with preserved goblet cell numbers in the ileum Protective and pathogenic effects of the IL- related cytokines in the SKG mouse model of... muc- expression Ileum Balb/c SKG + Anti IL- Ab Goblet cells in Ileum SKG ZAP- mutation Auto reactive T cells Fungal-B glucan Signalling through Dectin- Inflammatory cascade involving IL- /Th axis SpA Axial and peripheral IBD Arthritis and Ilieitis are IL- dependent... Day Day Day Day SKG D Balb/c D SKG D tx IL- is required for severe disease of joints and ileum and relatively protects the entheses IL- modestly supresses arthritis and plays a protective role in the gut Treatment with neutralising IL- is highly effective for both gut and joint IL- is likely from the gut SKG d Ileum no disease SKG d Ileum disease Anti-IL- relative enthesis sparing Anti-IL- enthesis sparing benefit unclear and potentially hazardous to gut ER stress is linked to IL- production in the ileum IL- drives both effector and inflammatory cytokines at initiation of disease and with progression IL- reduces goblet cell numbers and gene expression of muc- indicating a pathogenic role in mucous layer protection in the ileum

5 // Acknowledgements UQ Diamantina Ititute Prof Ranjeny Thomas Prof Matt Brown UQDI Thomas Lab SKG cluster group Linda Rehaume Jared Velasco Athan Baillet Michael Osbourne (BRF) MMRI Mike McGuckin Sumaira Hasnain Eli Lilly Kristine Kikly

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