In keeping with the effects on angiogenesis induced by contact hypersensitivity reactions in mouse ears, VS-I-treated mice revealed significantly reduced oedema formation, resulting from lower plasma leakage and inhibition of inflammation-associated vascular remodelling [66]. Intravital microscopy studies of inflamed ears showed a decrease

in the fraction of rolling leucocytes in VS-I-treated mice [66]. In addition to anti-microbial activity [67] Cgs may play important role in the neuroimmune interaction in relation to inflammatory function. This review will remain focused upon the function of Cgs in inflammatory responses in the gut. Circulating CgA levels, a marker for neuroendocrine tumours including carcinoids, have Protease Inhibitor Library recently been found elevated in some patients with IBD [68]. In this context the disease activity and TNF-α levels influence the CgA pattern, which could reflect the neuroendocrine system activation in

response to inflammation [69]. In a recent letter addressed to the aforementioned study, Sidhu and collaborators [70,71] confirmed the observation of Sciolia et al.[69] of an elevated level of CgA see more serum in both IBD and diarrhoea-predominant IBS patients. The unifying hypothesis proposed could be the EC cell hyperplasia producing an elevated serum CgA levels, as reported previously [72]. The differential replication of EC cells in IBS patients could also explain why elevated levels are found only in a proportion of patients, and levels decline with time. Further studies of serial serum CgA measurements in both these conditions would strengthen our understanding of the plausible mechanisms behind these observations. In the context of experimental colitis, intrarectal injection of CAT can decrease the inflammatory markers [73]. Disease activity index, macroscopic and histological scores, as well Erlotinib price as myeloperoxidase

(MPO) activity, were decreased significantly in mice treated with CAT compared to mice that received DSS only. Treatment decreased the onset of clinical disease as assessed by loose stools, weight loss and rectal bleeding. In addition, colonic tissue levels of IL-1β, IL-6 and TNF-α were decreased significantly in mice treated with CAT. Conversely, the biochemically modified fragment had no effect on the severity of colitis. These results support the hypothesis that Cgs-derived peptides modulate intestinal inflammation in a murine model of colitis by acting directly or indirectly on the microbiota and the immune system. Identification of the molecular and cellular mechanisms underlying the protective role of this peptide may lead to a novel therapeutic option in IBD.

Microsurgery 30:397–400, 2010. “
“Autologous breast reconstruction is safe in advanced age, yet no study has examined its effects on the aging abdomen. We, therefore, studied 145 women who participated in a prospective study of abdominal strength following abdominal free flap breast reconstruction, comparing preoperative and late follow-up scores JQ1 datasheet in patients ≥60 years old (11 unilateral, 13 bilateral) compared with patients <60 (58 unilateral, 63 bilateral). Simple in-office tests were utilized to test abdominal strength. No differences were noted in unilateral absolute scores at either time point, however, a decrease in upper abdominal strength was noted in the younger cohort over time (P = 0.01). Bilateral

analyses revealed absolute score decreases

in upper abdominal strength for both cohorts but no major differences between the two. We conclude that autologous breast reconstruction with abdominal tissue in older patients result in little to no difference in abdominal function as compared with younger patients. © 2012 Wiley Periodicals, Inc. Microsurgery, click here 2013. “
“Background: Large or extensive gouty tophi on the feet can cause functional impairment, drainage sinus, and infected necrosis, finally resulting in complex soft-tissue defects with tendon, joint, bone, nerve, and vessel exposure. Reconstruction of complex soft-tissue defects of the foot is still challenging. The purpose of this report was to review the outcomes of free-flap reconstructive surgery for treating the metatarsal joint defects of the feet caused by chronic tophaceous gout. Methods: Ten patients who had large tophus masses (>5 cm) and ulceration on the feet were admitted to our hospital between September 2006 and September 2010. Six patients underwent free-flap reconstruction after debridement to resurface the circumferential wound, protect the underlying structures, and provide a gliding surface for exposed tendons. The patients’ age, sex, comorbidities, location and size of the defects, reconstructive procedures,

surgical outcomes, complications, Janus kinase (JAK) follow-ups, and recurrence of tophaceous gout were reviewed and recorded. Results: The mean patient age was 49.8 years (range, 36–72 years). The average skin defect size was 92.2 cm2. Five patients were treated using free anterolateral thigh flaps, and 1, using a free medial sural flap. These free flaps were safely raised and showed excellent functional and cosmetic results, with a mean follow-up of 31.7 months (range, 7–50 months). Conclusion: Chronic tophaceous gout can cause severe skin infection and necrosis, even resulting in deformity or sepsis if left untreated. Surgical debridement is inevitable in patients with extensive wounds. We reconstructed the large, ulcerative skin and soft-tissue defects on the dorsum of the foot by performing free-flap reconstruction after adequate debridement and achieved good functional and cosmetic results. © C 2011 Wiley Periodicals, Inc. Microsurgery, 2011.

However, the EE-induced changes are not merely transcriptional and extend to effects on the proteome [126–128]. The cellular effects of EE, which are presumably

dependent on molecular changes, include enhanced adult neurogenesis [108,129–133] and synaptic plasticity [134–139]. Specific neuronal cell populations have been shown to be activated by EE [140] and the effects in the brain extend to glia [141–143]. However, a range of other cellular effects have been described, including those impacting on metabolism [144], the immune system [145–148] and the HPA-axis [47,149–151]. The EE-induced increase in adult hippocampal neurogenesis may contribute to enhancement of specific cognitive functions, in particular pattern separation [108,152–154], but is unlikely to be PLX-4720 datasheet sufficient for the broader behavioural benefits [155]. One

BIBW2992 in vivo key question that arises from EE studies is the extent to which the different components of EE (sensory stimulation, cognitive activity and physical exercise) can be separated and analysed with respect to their beneficial effects. The easiest aspect to assess separately, and the most studied, has been physical exercise. Laboratory mice and rats will voluntarily run long distances when provided with ad libitum access to running wheels. Whilst other forms of exercise, such as treadmill running have been used, those that require aversive stimuli to induce exercise are known to increase stress, which can confound such experiments. There is evidence that increased voluntary physical exercise (usually wheel

running) can enhance cognition and alter affective and motor states in wild-type rodents, and may induce at least some of the cellular changes associated with EE [5,7,156–158]. One idea which has been previously proposed is that mechanisms mediating the kinds of experience-dependent plasticity discussed above could be investigated for the development of ‘enviromimetics’, drugs which would mimic or enhance EE-induced therapeutic effects [159,160]. Enviromimetics could boost the beneficial effects of cognitive stimulation and physical activity. Physical exercise is known to contribute to many of the major effects of Tenofovir EE, such as increased adult hippocampal neurogenesis [5,156,161–163]. A more specific form of enviromimetic could thus be an ‘exercise mimetic’ that selectively enhances molecular and cellular processes induced by physical activity. So what might be an example of a well characterized molecular target for enviromimetic drugs? The most obvious example is BDNF, a neurotrophin whose expression is found to be induced by increased physical exercise [164], learning [165] and EE [124,125]. Furthermore, BDNF has been implicated in mechanisms of adult neurogenesis and synaptic plasticity, and thus is a key mediator of experience-dependent cellular plasticity in both the developing and adult nervous system [166].

In order to understand more clearly the gene transcriptional profiles associated Galunisertib research buy with CsA treatment in OS patients, 90 genes related to the immune system were examined by TLDA before and after successful treatment (patient

1). After treatment, 26·6% (24 of 90) of genes showed an expression level of more than twofold increase or decrease compared with the patient’s baseline gene expression (Fig. 4). Of these, the expression of 11 genes (12·2%) was down-regulated (by a factor of 2·3–5·2, values of 0·44–0·19, respectively, in Fig. 4, and 13 genes (14·4%) were up-regulated (by a factor of 2·04–19). The expression of several genes that are known to be down-regulated by CsA therapy such as IL-2 and Fas ligand this website (FasL) were found to be low (0·197- and 0·32-fold decrease). Interestingly, several genes that are known to be involved in immune regulation and autoimmunity were found to be markedly up-regulated

[e.g. IL-10, intercellular adhesion molecule (ICAM) 1 and transforming growth factor (TGF)-β] or down-regulated (e.g. CCR4 and CCR5). The immunological hallmark of OS is the expansion and activation of an oligoclonal population of autoreactive T cells. We have already shown that, in OS, similar T cell expansions are found in peripheral blood and in target organs (e.g. skin) [12]. These cells should be controlled rapidly by immunosuppressive agents to avoid tissue infiltration and to improve the general outcome of OS patients [14]. Here we describe a selective immune response to such treatments in patients with Omenn phenotype. Diverse topical and systemic immunosuppressive

therapies have been shown to be useful in OS patients. Many use CsA as the gold standard treatment for these patients. Alternatively, tacrolimus (FK506) is used. Despite similarity in their accepted mode of action, they alter T cell receptor expression differentially in vivo, therefore can have different effects on OS patients [15]. Failure of treatment HSP90 sometimes requires alternative or a combination of therapies [16]. Herein, we report on two patients; the first patient responded to CsA treatment while the second patient did not. Surprisingly, the initial expanded oligoclonal autoreactive clone (TCR-Vβ 17) in the latter patient responded well to CsA treatment, but other TCR-Vβs had started to expand, probably causing the patient’s unremitting autoimmune symptoms. Many unknown environmental and/or host factors can produce expanded lymphocytes in OS. In some cases the trigger (e.g. infection) that exacerbates the autoreactive process is found [17]. Patient 2 underwent a thorough infectious work-up, which was found to be negative, and no other obvious factor to trigger his symptoms could be detected to explain the presence of new TCR clones. However, expansion of certain new TCR-Vβ clones may also represent not only pathogen exposure, but also skewing towards self-antigens and autoimmunity [9].

The mRNA expression of Collagen IV, Fibronectin, TGF- β1 and MCP-1 in kidney tissue were lower in animals treated with PXS64 and Telmisartan (p < 0.05 vs UUO). In addition, mice treated with PXS64 had lower Collagen

IV, Fibronectin and phospho-Smad2 protein expression. PXS-64 inhibited latent TGFβ1-induced protein expression of collagen III, fibronectin and phospho -smad2 protein levels in HK-2 cells (P < 0.05 vs latent TGFβ1). Conclusion: Our data demonstrated that PSX64 significantly inhibited the effect of latent TGF β1on renal fibrotic and inflammatory Selleckchem Ipilimumab markers, suggesting that PSX64 is an effective agent in preventing kidney fibrosis. LEE SANG HO, KIM DONG-JIN, KIM SE YUN, SEO JEONG WOO, KIM YANG GYUN, MOON JU YOUNG, LEE ARAH, KIM MYUNG JAE, LEE TAE WON, IHM CHUN GYOO Division of Nephrology Department

of Internal medicine Kyung Hee University College of Medicine Introduction: Role of Bone marrow, a reservoir for endothelial AZD1208 price precursor cells (EPCs) and mesenchymal stem cells (MSCs), in kidney regeneration is still obscure. Recently substance-P (SP), an injury-inducible messenger to mobilize bone marrow stem cells, has been suggested to be a novel target of regenerative medicine. We investigated the long-term effects of the SP on kidney exposed to IRI. Methods: Unilateral renal ischemia–reperfusion injury (IRI) model was established in C57BL/6 mice and 5 ng/kg/day SP or saline was administered twice a week for 5 weeks after the surgery. Renal function was monitored, and histological changes and fibrosis in the kidney were evaluated in both two weeks and five weeks. TGF-β1 and α-SMA expressions, markers of renal fibrosis were determined by Western blot. The infiltration of macrophages in renal tissues was also assessed by immunohistochemistry. Results: Renal IRI increased SP levels in peripheral blood. Mobilized EPCs and MSCs in peripheral blood showed peak ID-8 at 1 day after IRI, followed by subsequently

decreased at 3 and 5 days. Administration of SP maintained the peripheral mobilization of EPCs to 5 days. Tubular injury scores of the SP group were significantly lower than those of the saline group at both two and five weeks. Interstitial fibrosis was also consistent with the result of tubular damage as there was significantly lower degree of interstitial fibrosis in SP group at 5 weeks. Intrarenal TGF-β1 and α-SMA expressions in SP treated group were significantly lower than those in saline treated group. Infiltration of macrophage was also significantly decreased in SP treated group. Conclusion: Our data show that long-term administration of SP ameliorates kidney damage and fibrosis after ischemic reperfusion injury and suggest the possible role of SP and bone marrow derived stem cells in kidney regeneration.

Some studies have reported that PI3K inhibition reduces Th2 cytokine production, pulmonary eosinophilia, airway inflammation, and bronchial hyperresponsiveness in a mouse asthma model 48, 49. In addition, PI3K is shown to be involved in HIF-1α activation induced by oxygen-dependent or oxygen-independent pathways 50, 51. Recently, p110δ and p110γ isoforms have sparked a great deal of interest, as there is increasing evidence that Decitabine cost these isoforms play key roles in immunity 52. We have also demonstrated that OVA-induced HIF-1α

activation is significantly reduced by administration of a PI3K-δ inhibitor and suggested that inhibition of the p110δ signaling pathway has therapeutic potential for allergic airway inflammation 33. Consistent with these observations, in the present study, levels of p-Akt protein and PI3K activity in lung tissues were increased after OVA inhalation. The increased levels of p-Akt and PI3K Nutlin-3 nmr activity were significantly reduced after administration of IC87114, a PI3K-δ inhibitor. Moreover, the increased levels of HIF-1α after OVA inhalation were significantly reduced by IC87114 in primary tracheal epithelial cells isolated from OVA-treated mice. These findings suggest

that PI3K-δ regulates HIF-1α activation therewith inducing VEGF expression in a murine model of allergic airway disease. In summary, we have examined the roles of HIF-1α in allergen-induced airway inflammation and bronchial hyperresponsiveness using an HIF-1α inhibitor,

2ME2, and siRNA targeting HIF-1α and evaluated the role of PI3K-δ signaling in HIF-1α activation in allergic airway disease. The administration of 2ME2 was effective in reversing all pathophysiological symptoms examined. Our data have also revealed that HIF-1α inhibition substantially reduces the increase in VEGF expression as well as the activity of VEGF in lungs, especially in bronchial epithelial cells, of our murine model of allergic Sorafenib mouse airway disease. In addition, administration of IC87114 reduced the increase in HIF-1α activity in OVA-treated airway epithelial cells. Therefore, one likely mechanism for the roles of HIF-1α in pathobiology of allergen-induced airway inflammation and hyperresponsiveness is induction of vascular leakage via up-regulation of VEGF expression in lungs as well as bronchial epithelium. Thus, these findings provide a crucial molecular mechanism for the potential of HIF-1α inhibition in preventing and/or treating asthma and other airway inflammatory disorders. Female C57BL/6 mice, aged 8–10 wk and free of murine specific pathogens, were obtained from the Orientbio (Seoungnam, Korea) were housed throughout the experiments in a laminar flow cabinet and were maintained on standard laboratory chow ad libitum.

Cultures were collected from the different anatomical sites of all the patients within 24 h of diagnosis of candidemia. Molecular similarities between identical species colonised with Candida species were evaluated via karyotyping. The colonisation index, as developed by Pittet et al. was calculated using screening culture results from patients. Among the 40 patients screened for colonisation, 35 (87.5%) had colonisation of at least one anatomical site. Twenty-six (74.3%) of the 35 patients with colonisation in any of the three anatomical sites (respiratory, rectum and urinary sites) were shown to be colonised with the same species that caused candidemia. When the anatomical sites

were compared with each other, no significant difference was observed at the species level in terms of colonisation index. The colonisation index (≥0.5) positivity rate was 74% in patients with candidemia. www.selleckchem.com/products/LDE225(NVP-LDE225).html Selleckchem AT9283 The investigation of Candida colonisation of at least three anatomical (respiratory, rectum and urinary) sites could

help in the selection of empirical antifungal therapy when nosocomial candidemia is suspected. “
“Pulmonary coccidioidomycosis is caused by inhaling airborne arthroconidia of Coccidioides, a soil-dwelling fungus endemic to the desert southwestern United States. Although uncommon, disseminated coccidioidal infection can be associated with well-defined risk factors, such as cell-mediated immunodeficiency, certain racial heritages (e.g. African or Filipino), male sex, or pregnancy. Before widespread use of computed tomography (CT), the presence or persistence of mediastinal lymphadenopathy was postulated to be a risk factor for disseminated coccidioidal infection. To investigate the use of CT scanning to identify the presence of mediastinal lymphadenopathy in patients Protein kinase N1 with pulmonary coccidioidomycosis, and to correlate such lymphadenopathy with disseminated coccidioidal infection, we performed a retrospective review of patients with pulmonary coccidioidomycosis who were

evaluated by chest CT. Two radiologists independently interpreted 150 CT scans from patients with pulmonary coccidioidomycosis. Forty-nine patients met CT criteria for mediastinal lymphadenopathy, whereas 101 patients did not. Disseminated coccidioidal infection was observed in 5 (10%) of the 49 patients with mediastinal lymphadenopathy and in 6 of the 101 (6%; P = .34) without such adenopathy. Among patients with coccidioidomycosis, patients with mediastinal lymphadenopathy, as assessed by CT, had a higher rate of disseminated infection, but the difference was not statistically significant. “
“The results of the use of ozonised sunflower oil (OLEOZON®) in the treatment of onychomycosis, based on its known antimycotic action and good skin tolerance, by means of a controlled randomised phase III assay are presented.

After disruption by incubation at 37°C for 30 min in HBSS (Invitrogen) containing 0.5 mg/mL collagenase D (Roche), DCs were purified by magnetic separation using anti-CD11c MACS microbeads. Non-specific binding was blocked using unlabeled anti-FcγR (BD Biosciences). Cell purity was assessed by flow cytometry and always greater than 92%. For P3C cultures, CD4+CD25+ T cells purified from naïve female NOD mice were cultured for 6 days with 2 μg/mL P3C and DCs purifed from naïve female NOD mice, at a ratio of 1 DC:3 Tregs, in RPMI 1640 supplemented with 10% FCS, 2 mM L-glutamine, click here and 50 μM 2-mercaptoethanol (Complete RPMI), and 10 U/mL rhIL-2. For viral cultures, the CD4+CD25+ T cells were purified from female B6 mice

infected 21 days prior with LCMV and cultured for 6 days with DCs purifed from female B6 mice infected 48 h prior with LCMV, at a ratio of 1 DC:3 Tregs, in Complete RPMI. At the end of the cultures, the Deforolimus cost Tregs were negatively selected using rat anti-mouse MHC class II mAbs (BD Biosciences) and Sheep anti-rat Dynabeads

(Dynal). Statistical significance was determined using a logrank test (for T1D assessment) or an unpaired, two-tailed t-test. In all experiments, differences were considered significant when p<0.05. Statistical significance is displayed in each figure for the indicated groups as follows: *p<0.05, **p<0.005, ***p<0.001. The authors thank Malina McClure for mouse colony maintenance, Yang Chen and Tom Wolfe for technical help, and Priscilla Colby for administrative assistance. This work was supported by an NIH P01 grant AI58105-03 with the NIAID for M.G.vH, and fellowships from the JDRF and FRM for C.M.F. The authors also gratefully acknowledge support from the Brehm Coalition. Conflict of interest: The authors declare no financial or commercial conflict BCKDHB of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”.

Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. “
“During chronic lung infection of patients with cystic fibrosis, Pseudomonas aeruginosa can survive for long periods of time under the challenging selective pressure imposed by the immune system and antibiotic treatment as a result of its biofilm mode of growth and adaptive evolution mediated by genetic variation. Mucoidy, hypermutability and acquirement of mutational antibiotic resistance are important adaptive phenotypes that are selected during chronic P. aeruginosa infection. This review dicsusses the role played by these phenotypes for the tolerance of biofilms to antibiotics and show that mucoidy and hypermutability change the architecture of in vitro formed biofilms and lead to increase tolerance to antibiotics. Production of high levels of beta-lactamase impairs penetration of beta-lactam antibiotics due to inactivation of the antibiotic.

WU HUNG-LIEN1,3, SUNG JUNNE-MING2, TSENG CHIN-CHUNG3, WANG MING-CHENG4 1Department of Nutrition, National Cheng Kung University Hospital, Tainan; Taiwan; 2Internal Medicine, National Cheng Kung University Hospital, Tainan; Taiwan; 3Internal Medicine, National Cheng

Kung University Hospital, Tainan; Taiwan; 4Internal Medicin, National Cheng Kung University Hospital, Tainan; Taiwan Introduction: The subjective global assessment (SGA) is a good nutritional assessment method and predict the outcome in dialysis patients, but fewer studies analysis the 6 items in SGA to effect on the outcome of patients with chronic peritoneal dialysis (CPD). The purposes of the study investigate the 6 items from SGA affected outcome of CPD patients selleck in Southern PD0325901 concentration Taiwan. Methods: Our study enrolled 183 chronic PD

patients (92 males and 91 females) from National Cheng Kung University Hospital, Tainan, Taiwan and new CPD patients from 2003 to 2012, and fellow up 9 years. For assessment of nutritional status used a 7 point of SGA scales, the method include six items, as weight loss in the preceding 6 months, appetite, gastrointestinal symptoms, daily activity, disease stress, and the physical examination. Results: Older, DM, cancer, CAD, hyperlipidemia, and before PD received HD patients had higher dropout rate. Higher total SGA score, appetite score, GI function score, activity score had better outcome. Univariated Cox’s regression model Ibrutinib mw analysis for reaching end points in CPD patients: age (HR (95% CI): 1.03 (1.02–1.05), P < 0.001),

Cancer (HR (95% CI): 2.17 (1.12–5.10), P = 0.022), DM (HR (95% CI): 2.15 (1.28–3.62), P = 0.004), CAD (HR (95% CI): 2.28 (1.26–4.12), P = 0.006) were higher risk, but higher total SGA score (HR (95% CI): 0.78 (0.64–0.95), P = 0.017), body weight change score (HR (95% CI): 0.82 (0.69–0.98), P = 0.028), GI function score (HR: 0.77 (0.65–0.92), P = 0.003), activity score (HR: 0.72 (0.61–0.86), P < 0.001) can significantly decrease the risk of dropout from CPD. Conclusion: older age, DM, and CAD increase the risks, but higher total SGA score, especially higher activity score can reduced hazard ratio and increase outcome in CPD patients. ROJSANGA PIYARAT Dialysis unit, Medicine Department, Udon Thani Hospital, Thailand Introduction: Continuous ambulatory peritoneal dialysis (CAPD) is the main renal replacement therapy (RRT) in Thailand due to universal coverage scheme. CAPD associated peritonitis is the major complication in CAPD. From previous studies showed that advanced age, diabetes, high body mass index, hypoalbuminemia and high blood sugar were associated with increase in incidence of CAPD associated peritonitis. This study was conducted to evaluate the risk factors of peritonitis in CAPD clinic in Udon Thani Hospital.

The resulting cell suspensions were re-suspended in F-12 Nutrient mixture (Gibco-Invitrogen) mixed 1:1 with DMEM supplemented with 10% FCS, 1% L-glutamine, 1% penicillin/streptomycin, 1% HEPES and 1% non-essential amino acids. The cultured cells were allowed to form colonies in 6-well tissue culture plates (Nunc-Fisher Scientific) for 7 days, then lifted using 0.2% Na2EDTA, reseeded into T75 flasks at 1×106/flask and cultured for a further 7 days before use

in co-culture experiments. Single cell suspensions were prepared from mouse spleen and lymph nodes by mechanical disruption and filtering through 150 μM Sefar Nitex ribbon mesh (Sefar, Lancashire, UK) followed by erythrocyte lysis in ACK lysis buffer for 3 min at room temperature. Cell click here suspensions were incubated with anti-mouse CD4 microbeads (Miltenyi Biotec, Auburn, CA, USA) for 20 min at 4°C, washed in MACS buffer and separated MI-503 in vivo using MS columns and an OctoMACS® separator according to the manufacturer’s instructions (Miltenyi Biotec). CD4+ fractions were washed in MACS buffer, re-suspended

in culture medium and used as responders in activation cultures. CD4− fractions were depleted of remaining T cells using anti-CD90.2 microbeads by the same protocol and were used as APCs. For Th17 differentiation, CD4+ T cells and APCs were cultured for 4 days in 96-well round bottom plates (Sarstedt, Nümbrecht, Germany) Ribonuclease T1 or for 3 days in the lower compartment of Corning® HTS Transwell® 9-well permeable supports (Sigma-Aldrich) at 1×106/mL and 2×106/mL respectively with 1 μg/mL anti-CD3ε, 5 μg/mL anti-IFN-γ, 4 μg/mL anti-IL-4, 5 ng/mL TGF-β1 and 25 ng/mL IL-6. In some experiments, CD4+ T cells were cultured at 1×106/mL with 1:1 Dynabeads®. Other reagents were added as described for individual experiments. For all co-culture experiments, MSCs or fibroblasts were re-suspended in DMEM/10% FCS, added in graded numbers to the wells of 96-well round bottom plates and allowed to adhere for 4 h prior to the addition of CD4+ T cells/APCs or CD4+ T cells/Dynabeads®.

For re-stimulation of Th17-skewed T cells from primary cultures and co-cultures, cells were subjected to magnetic separation using anti-CD4 microbeads with positive column fractions saved. The resulting re-purified CD4+ T cells were re-plated at 0.5×106/mL in fresh medium containing 1:1 Dynabeads® with no other additions in 96-well round bottom plates for a further 24 h. For some experiments, CD4+ T cells were labelled for analysis of proliferation by flow cytometry using CellTrace CFSE cell proliferation kit (Molecular Probes®, Invitrogen). Supernatants from cultures and co-cultures were analysed by ELISA using DuoSet® ELISA Development Systems (R&D Systems, Minneapolis, MN, USA) for IL-17A and IFN-γ and a Parameter Assay Kit for PGE2 (R&D Systems). For flow cytometry, cells were suspended in FACS buffer at 5.