The sequences of the primers were as follows: 5′-AGGGTAGTTAGTTTTC

The sequences of the primers were as follows: 5′-AGGGTAGTTAGTTTTCGGAAC-3′

(forward) and 5′-CCATTAACGTCATAACGACC-3′ (reverse). The primers for the internal reference gene β-actin were designed to amplify the region that is devoid of CpG nucleotides. The β-actin primer sequences were 5′-TGGTGATGGAGGAGGTTTAGTAAGT-3′ (forward) and 5′-AACCAATAAAACCTACTCCTCCCTTAA-3′ (reverse) 60. Relative FOXP3 methylation levels of different T cells were normalized to β-actin gene expression and compared with the expression level of methylated FOXP3 in CD4+CD25- T cells (set as 100%). All experiments were performed in triplicate. Total RNA was extracted from T cells using Trizol reagent (Invitrogen), MAPK Inhibitor Library cost and cDNA was transcribed using a SuperScript II RT kit (Invitrogen), both according to the manufacturers’ instructions. TCR-Vβ mRNA expression was determined by RT-PCR using specific 29 pair primers, and check details mRNA levels in each sample

were normalized to the relative quantity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), as previously described 29, 30. Transcription factor, cytokine and receptor expression were analyzed using real-time quantitative PCR 35, 61. Relative mRNA expression was calculated using the comparative method for relative quantification following normalization to GAPDH gene expression. All experiments were performed in triplicate. The specific primers used are listed as follows: Unless indicated otherwise, data are expressed as means±standard deviation (SD). Paired or unpaired two-tailed Student’s t-test was used to analyze differences between two groups. Differences were considered significant for p-values <0.05. The authors thank Chris Eickhoff for technical assistance.

This work was partially supported by a grant from the American Cancer Society (to G. P) and a seed grant (to G. P) from the Cancer Center at Saint Louis University. Conflict of interest: The authors declare no financial or commercial Amine dehydrogenase conflict 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. “
“Microglia are the major myeloid-immune cells of the brain parenchyma. In a steady state, microglia monitor their environment for pathogens or damaged cells. In response to neural injury or inflammation, microglia become competent APCs able to prime CD4+ and CD8+ T lymphocytes. We previously demonstrated that neonatal and adult microglia cross-present exogenous soluble Ags in vitro. However, whether microglia are able to cross-present Ag to naive CD8+ T cells in vivo, within the brain microenvironment, remains undetermined. Here, we have designed an original protocol in order to exclude the involvement in cross-presentation activity of peripheral migrating APCs and of CNS-associated APCs.

The placental vascular dysfunction does extend to other fetal vas

The placental vascular dysfunction does extend to other fetal vascular beds including endothelial cells from umbilical vessels, where there are reports of elevated basal iNOS activity and altered sensitivity to insulin. There is emerging evidence of epigenetic modulation of fetal endothelial Galunisertib nmr genes in diabetes and long-term vascular consequences

of this. Thus, placental vascular dysfunction in diabetes may be contributing to and describing disturbances in the fetal vasculature, which may produce an overt pathological response in later life if challenged with additional cardiovascular stresses. “
“Please cite this paper as: Arkill KP, Neal CR, Mantell JM, Michel CC, Qvortrup K, Rostgaard J, Bates DO, Knupp C, Squire JM. 3D reconstruction of the glycocalyx structure in mammalian capillaries using electron tomography. Microcirculation 19: 343–351, 2012. Objective:  Visualising the molecular strands making up the glycocalyx in the lumen of small blood vessels has proved to be difficult using conventional transmission electron microscopy techniques. Images obtained from tissue stained in a variety of ways have revealed a regularity in the organisation of the proteoglycan components of the glycocalyx layer (fundamental spacing about 20 nm), but require a large sample number. Attempts to visualise the glycocalyx face-on (i.e. in a direction perpendicular to the endothelial cell layer in the Alectinib research buy lumen and directly

applicable for permeability modelling) has had limited success (e.g. freeze fracture). A new approach is therefore needed. Methods:  Here we demonstrate the effectiveness of using the relatively novel electron microscopy technique of 3D electron tomography on two differently stained glycocalyx preparations. A tannic acid staining

method and a novel staining technique using Lanthanum Dysprosium Glycosamino Glycan adhesion (the LaDy GAGa method). Results:  3D electron tomography reveals details of the architecture of the glycocalyx just above the endothelial cell layer. The LaDy GAGa method visually appears to show more complete coverage and more depth than the Tannic Acid staining method. Conclusion:  The tomographic reconstructions show a potentially significant improvement in determining N-acetylglucosamine-1-phosphate transferase glycocalyx structure over standard transmission electron microscopy. “
“Please cite this paper as: Ella, Yang, Clifford, Gulia, Dora, Meininger, Davis and Hill (2010). Development of an Image-Based System for Measurement of Membrane Potential, Intracellular Ca2+ and Contraction in Arteriolar Smooth Muscle Cells. Microcirculation17(8), 629–640. Objective:  Changes in smooth muscle cell (SMC) membrane potential (Em) are critical to vasomotor responses. As a fluorescent indicator approach would lessen limitations of glass electrodes in contracting preparations, we aimed to develop a Forster (or fluorescence) resonance energy transfer (FRET)-based measurement for Em.

8% However, the pooled incidence of AKI requiring RRT remained l

8%. However, the pooled incidence of AKI requiring RRT remained largely unaffected (pooled crude incidence, 0.86%). The increase of the pooled AKI incidence may reflect that AKIN and RIFLE criteria were the most sensitive diagnostic criteria for AKI among our studies. Besides, the study included patients undergoing noncardiac surgery[46] had the lowest check details crude incidence

of AKI, among all the seven studies using AKIN and RIFLE criteria. These findings pointed out the impact of surgery type and diagnostic definition of AKI when considering the incidence of AKI. Importantly, since RIFLE and AKIN criteria have become the mainstays of diagnostic definition for AKI, caution should be exercised when it comes to interpret the past studies not applying these criteria for diagnosis. The strength of our meta-analysis and systemic review include the comprehensive search, the large sample size, the inclusion of latest studies with high methodological quality, multiple subgroup analyses, and low statistical heterogeneity with regards to the outcome of postoperative AKI requiring RRT. Our study also provided a review of the incidence of postoperative AKI and postoperative AKI requiring RRT in the context of the specific type of surgery and specific definition of AKI (Table 1). There were

several limitations of our study. As with all the observational studies, the causal relationship was hard to establish and there might be unknown confounders left unadjusted even after meticulous ifoxetine search for confounders. buy Decitabine Besides, the variation in types of surgery, the heterogeneity of the definition of postoperative

AKI, and the lack of the complete report of preoperative statin therapy were also problems. Different types of surgery pose different risk on postoperative AKI. In cardiac surgery, duration of CPB may be an important risk factor for AKI,[56] but this information was not provided in most studies. In other major surgeries other than cardiac surgery, the pathophysiology of renal insult is not as clear. The intensity of surgery-related insult to the kidney in different types of surgery may vary, and this effect was unable to be adjusted for. The level of emergency of the operation might also influence the risk of AKI, but this information was also unavailable for our meta-analysis. Although a dose dependent renoprotective effect was demonstrated in two studies,[43, 57] the majority of studies did not report the specific type, dosage, and duration of preoperative statin therapy. In studies reporting the detail of preoperative statin therapy, the specific type, dosage, and duration of statin therapy were often not uniform among studies. In chronic statin users, early re-institution of statin therapy after operation might be beneficial, but only one study[38] reported outcome relevant to this kind of statin exposure.

0 software

The difference was considered statistically s

0 software.

The difference was considered statistically significant when P ≤ 0.05. Leica Microscopy system was used to take the picture, and magnification used was 40 with numerical aperture of the objectives, at temperature room. The slides were mounted using Vectashield mounting medium (Vector laboratories), and Alexa 488 fluorochrome was used to detect the positive signal (Invitrogen). As a first step, we designed recombinant adenovirus vectors containing ESAT-6 with and without calreticulin to determine whether calreticulin increased the immune response to the antigen. AdESAT-6 and AdCRT–ESAT-6 were created as described in the Materials and methods. Expression of ESAT-6 in both constructs was under the control of a cytomegalovirus promoter (Fig. 1A–C). The capacity of these constructs to express ESAT-6 was first verified by immunoblot Panobinostat in vitro analyses of HEK293 cells transfected with one of the recombinant vectors (data not shown). ESAT-6 protein expression was also demonstrated by immunofluorescence analysis of HEK293 cells transfected with AdESAT-6, AdCRT-ESAT-6 or AdLacZ (Fig. 1D). As shown in Fig. 1D, only cells transfected with

AdESAT-6 and AdCRT-ESAT-6 express ESAT-6. Therefore, our recombinant adenovirus constructs were proven to be capable of producing ESAT-6. To test the ability of AdCRT–ESAT-6 or AdESAT-6 to generate ESAT-6-specific cellular immune responses in vivo, mice PLX4032 cost were immunized by the intranasal route with the adenovirus constructs.

At 4 weeks post-vaccination, splenocyte cultures were prepared Thalidomide and restimulated with ESAT-6, and the resultant cytokine responses were analysed. It was found that while splenocytes from mice immunized with the antigen alone (AdESAT-6) showed no differences in cytokine production compared to splenocytes from LacZ-immunized mice (controls), there were significant inductions of IFN-γ and TNF-α (measured by ELISPOT and ELISA, respectively) in splenocytes from mice immunized with the antigen ESAT-6 fused to calreticulin (AdCRT–ESAT-6) (Fig. 2A,B). Taken together, these data demonstrate that immunization with ESAT-6 linked to calreticulin is an effective approach to generate potent immune responses. It has been previously shown that fusion of ESTA-6 with CFP-10 enhances the immune response. Hence, using the same strategy, we expressed a calreticulin–ESAT-6–CFP10 fusion protein (AdCRT–ESAT-6–CFP10) and compared its ability to induce a cytokine response against AdCRT–ESAT-6. The expression of the fusion protein was demonstrated by immunoblot analysis of lysates of cells transfected with the fusion vector using an anti-CFP10 polyclonal mouse antibody (Fig. 3A). While no reaction was observed in the uninfected HEK293 cell lysates, a single antibody-reactive band of approximately 90 kDa was detected in the AdCRT–ESAT-6–CFP10 cell lysates. The size of the reactive band correlated with the predicted size of the CRT-ESAT-6–CFP10 fusion protein.

In contrast, the overall immature phenotype of APC containing hig

In contrast, the overall immature phenotype of APC containing higher frequencies of subpopulations with regulatory or suppressive properties may render younger mice largely incapable of generating encephalitogenic T cells and may further protect them by promoting development of Th2 cells and Treg cells. In this study, we demonstrate that the animal model of MS, EAE, cannot be induced with a standard protocol in otherwise susceptible mice that are below a certain age. Disease resistance in younger mice was associated with a higher frequency of plasmacytoid DCs and myeloid-derived suppressor cells, two APC subtypes with immunosuppressive

properties [14, 17]. Furthermore, APCs from younger mice displayed a functionally immature phenotype characterized by a decreased expression of MHC II and co-stimulatory CD40, a reduced production of proinflammatory TNF, IL-6, IL-23, and IL-12 and an enhanced release of anti-inflammatory IL-10. These APCs were incapable of generating encephalitogenic T cells and promoted development of Treg-cell populations instead. As adoptive transfer of adult APC restored inducibility of EAE in young mice, we propose that during development the innate immune cell compartment may gradually shift from regulatory/suppressive properties to proinflammatory

function, which may represent one immunological factor that facilitates susceptibility to CNS autoimmune disease. Our results hence favor an age-related decline of regulatory APC phenotypes and myeloid derived suppressor cells and an increase in the expression of constitutive and inducible MHC II and co-stimulatory molecules on myeloid APCs and B cells Hydroxychloroquine mouse as explanation why young mice are protected from T-cell-mediated CNS autoimmune disease. It is clear that overall MHC II expression is required for initiation of EAE, as mice genetically engineered to lack MHC II molecules

are resistant to development of CNS autoimmune disease [21]. Further, it has been demonstrated that the density of MHC II-Ag complexes and thereby buy RG7420 the strength of TCR signaling can determine the fate of the corresponding T cell [22]. While a strong interaction between APCs and T cells was required to generate proinflammatory T cells, a weaker molecular contact triggered development of an anti-inflammatory T-cell response [23]. Besides sufficient stimulation via MHC II, CD40-CD40-L ligation is critical to further stabilize the APC-T-cell interaction after Ag recognition [24]. In vivo disruption of CD40-CD40-L interaction via a monoclonal anti-CD40L Ab completely prevented the development of EAE [25], suggesting that cross-ligation via CD40 is a requirement for effector T-cell development. In context with our new findings, these data further consolidate the conclusion that younger mice are protected from CNS auto-immune disease as lower expression levels of MHC II and CD40 on APCs may not suffice to generate encephalitogenic Th1 and Th17 effector T cells.

The interaction of IL-22 and TNF-α is mediated through the IL-22R

The interaction of IL-22 and TNF-α is mediated through the IL-22R heterodimer and tumor necrosis factor receptor I 26 and intracellularly by MAP kinases, in particular p38, which leads to downstream activation of AP-1 family transcription factors. The combination of IL-22 and TNF-α strongly induced the phosphorylation and translocation of MAP kinases to the nucleus whereas the single cytokines only weakly contributed to MAP kinase activation. It is known that both IL-22 27 and TNF-α 28 activate MAP kinases; however, main signaling pathways for IL-22 are mediated through the transcription factor STAT-3 and other STAT molecules 6, 24, while TNF-α strongly

induces the NF-κB signaling cascade in keratinocytes 29. Since NF-κB is not synergistically activated by the combination of

SB203580 price TNF-α and IL-22, the observed synergism does not cover the whole functional spectra of TNF-α and IL-22, LDE225 order but is rather limited to aspects such as innate immunity. This may explain functional diversity of TNF-α and IL-22 as well as a dual role for IL-22: alone it has protective effects and enhances wound healing 30, in combination with TNF-α it becomes immune-stimulatory and arms epithelia for innate responses. The stimulation of the epithelial immune system by the IL-22/TNF-α axis is important for defense against extracellular pathogens like C. albicans. Supernatant of keratinocytes pre-incubated with the combination of both cytokines or Th22 clone supernatant most effectively reduced Bay 11-7085 C. albicans growth, protected keratinocytes from apoptosis and conserved the epidermal structure in an in vitro Candida infection model. Interestingly, common side effects of an anti-TNF-α therapy (Infliximab) are serious respiratory and skin infections 31, which could be explained by the missing interaction of IL-22 with TNF-α. Therefore, the IL-22/TNF-α axis itself is protective and important for the homeostasis of the human organism and its environment; if not tightly

regulated, however, this strong synergism might turn pathologic and cause severe and chronic inflammatory skin diseases like psoriasis. In summary, the discovery of the IL-22/TNF-α axis as an essential combinatorial key for cutaneous immunity gives a first insight into the function of Th22 cells and could lead to new therapeutic approaches of chronic inflammatory skin diseases like atopic eczema and psoriasis. Primary human keratinocytes were obtained from human foreskin (Western blot analysis) or healthy adult volunteers (n=10). Before samples were taken, each participant gave his informed consent. The study was approved by the ethical committee of the Technical University Munich and was conducted according to the declaration of Helsinki. Keratinocytes were isolated using the method of suction blister as described previously 32. Briefly, blisters were induced by generating a vacuum on normal skin of the forearms. Epidermal sheets were obtained from blister roofs, treated with 0.

0 mg/day However, urine protein further increased beyond 1 g/g C

0 mg/day. However, urine protein further increased beyond 1 g/g Cr, and serum creatinine increased and C-reactive protein also increased, accompanied by skin rash and dyspnoea. Allograft biopsy was conducted (2.5 years post transplant). The biopsy showed diffuse glomerular endocapillary proliferation and swelling of glomerular endothelial cells (Fig. 1). The presence of glomerular basement

membrane injury was present. Immunofluorescence showed no significant immune deposit including C4d. Intraluminal proliferating cells were mostly CD34+, indicating that the majority of them were endothelial cells. On the contrary, CD4+ or CD8+, i.e. T cells or CD68+ macrophages were few by immunohistochemistry. These findings suggested that the injury was mediated by direct insult to the endothelium, such as drug-induced injury, and was not mediated by alloantigen-directed immunological insult. No endarteritis or tubulointerstitial lesion was observed. No donor-specific antibody was detected with flow-bead analysis. EVR was discontinued and TACER was returned to the previous dose. selleck Proteinuria only decreased to 0.5 g/g Cr and methylprednisolone mini-pulse therapy was given (125 mg ×3), resulting in improvement of proteinuria to 0.1 g/g Cr. Other symptoms have also disappeared. Allograft biopsy taken 6 months later still showed mild and focal endocapillary

proliferation but those lesions had significantly improved compared with the previous biopsy (Fig. 2). Glomerular injury accompanied by de novo proteinuria in this case is assumed to be caused by everolimus. The presence of glomerulitis supports antibody-mediated rejection (AMR) as a possible cause. However, glomerular endothelial injury was not associated with either lymphocyte margination or C4d deposition and the proliferating cells were mostly endothelial cells, indicating the injury was mediated by drug rather than alloimmune response. Furthermore, the lack of donor-specific antibody and the reversal of clinical and pathological findings only by low-dose steroid therapy are unsupportive of

AMR. The presence of basement membrane injury could be mediated by Mirabegron CNI toxicity. In this case, the deterioration of the clinical data occurred after adding EVR. The presence of underlying CNI-mediated glomerular injury could not be excluded but the injury, severe enough to induce significant clinical presentation was likely to be triggered by EVR. Typically, proteinuria induced by mTORi is believed to be mainly mediated by podocyte injury.[5] Reports of glomerulonephritis induced by EVR, as in our case, are scarce.[6] Reluctance to biopsy would have resulted in attributing the cause of proteinuria to typical adverse effect of EVR in general. We could fortunately reverse the graft injury after recognizing the presence of glomerulonephritis and this case suggests the importance of clarifying the pathology by allograft biopsy.

We have recently shown that the transplantation of BM transduced

We have recently shown that the transplantation of BM transduced with pMog promotes deletional tolerance and prevents development of the MOG35–55-induced EAE in C57BL/6 mice 29. Given that the ectopic expression of AIRE can induce expression of TRA, including AZD9291 MOG in vitro, we asked whether the transplantation of retrovirally transduced BM cells expressing AIRE in syngeneic animals altered the course of EAE in animals immunized with MOG35–55. The level of chimerism was analysed 10 weeks following the transplantation of transduced BM cells by assessing the percentage

of GFP+ cells from the thymus and spleen. The GFP expression was detected in all the major cell lineages examined, including

CD4+and CD8+ T cells, B cells and MHC class II+ CD11c+ dendritic cells (Fig. 3A, Supporting Information Fig. 1 for gating strategy). RT-PCR analysis of thymus samples from Aire chimeric mice revealed increased levels of Aire, Mog and Ins2 mRNA compared with thymi from mice transplanted with normal BM or from untouched WT mice, suggesting that the AIRE expression see more has upmodulated these two defined autoantigens (Fig. 3B). While attempted, we were not able to accurately quantify and compare the MOG expression in the thymus across normal mice, mice transplanted with normal BM or Aire-transduced BM. To demonstrate differential expression of Aire and TRA in cells originating from transduced BM cells, GFP+ cells were enriched from the spleens of chimeric mice. Comparison Avelestat (AZD9668) of GFP+ and GFP- cells indicated a greater level of AIRE expression in GFP+ cells, consistent with retroviral promoter-driven expression within these cells. Further analysis revealed elevated levels of Mog and Ins2 mRNA in GFP+ cells compared with GFP- cells (Fig. 3C). These data support our in vitro findings that the ectopic expression of AIRE can promote the expression of TRA including the autoantigens Mog and Ins2. We next determined whether the intrathymic expression of the EAE/MS associated autoantigens

Mog, Plp and Mbp was AIRE dependent. MHCIIhi mTEC (CD45–, Ly51–, MHCIIhi) from WT and Aire−/− C57BL/6 mice were isolated and qRT-PCR revealed a marked reduction in the expression of MOG (to 25% WT levels) and Plp (to 12% WT levels) in Aire−/− mTEC with no change in Mbp expression (Fig. 4A). While Mog has previously been reported as being AIRE dependent, PLP was reported to be AIRE independent 39. However, these data came from human association studies of AIRE and TRA expression rather than from the examination of AIRE-deficient thymi and could thus explain the discrepancy in result for PLP. Given the observed reduction in Mog expression, we asked whether Aire−/− mice were more susceptible to MOG-induced EAE than WT C57BL/6 mice.

After 2 h of incubation at room temperature, bound biotinylated p

After 2 h of incubation at room temperature, bound biotinylated peptide-MHC complexes were detected colorimetrically, as indicated

previously 31. TEPITOPE is a T-cell epitope prediction software that enables the identification of ligands binding in a promiscuous or allele-specific mode to HLA-DR molecules 32. We set the TEPITOPE prediction threshold to 10% to select all potential peptide binders to HLA-DR*0401, DR*0404, DR*0101 (which includes weak putative binders and may yield false positives, i.e. peptides not able to bind to these alleles; see 32). HLA-DR*0401-Tg mice were previously described 33 and were purchased from Taconics Farm (Germantown, NY, USA). Mice BYL719 datasheet were bred and kept under specific pathogen-free conditions. Mice were immunized subcutaneously with 3 nmole hnRNP-A2 peptide emulsified in CFA containing H37Ra Mycobacteria (Difco, Becton Dickinson). Eight days later, cells from the draining lymph nodes were cultured (6×105 cells/well) in 96-well culture plates (Costar) with 30 μM of Alectinib nmr hnRNP-A2 peptides in synthetic HL-1 medium (BioWhittaker) supplemented with 2 mM L-glutamine and 50 μg/mL gentamicin (Sigma). PPD was used

as positive control for each culture at a final concentration of 10 μg/mL. Cultures were incubated at 37°C and supernatants were harvested after 20 h (for the detection of IL-2) or 60 h (for the detection of IFN-γ). All animal procedures were performed according to Austrian laws (BGBI. I Nr 162/2005) and approved by the local ethical committee. IFN-γ and

IL-2 were detected by ELISA as previously described 34, using for capture anti-IFN-γ mAb selleckchem Ph551216 (PharMingen) or anti-IL-2 mAb JES6-1A12 (PharMingen), and for detection biotin-conjugated AN 18.17.24 mAb (kindly provided by Dr. Edgar Schmidt, Mainz) or biotinylated anti-IL-2 mAb JES6-5H4. The cytokine detection limit was 30 pg/mL. ELISPOT plates (Millipore Multiscreen-HA MAIP), pre-incubated with 70% ethanol and washed with distilled water, were coated overnight at 4°C with 10 μg/mL capture mAb (anti-IFN-γ mAb, BMS228ESCA/1, Bender Med Systems, Vienna, Austria) dissolved in 0.1 M NaHCO3 pH 9.5. The plates were then washed once with 200 μL sterile PBS and blocked with 200 μL/well complete RPMI medium (RPMI 1640, 1 mM sodium pyruvate, 200 μM L glutamine, MEM with nonessential amino acids, 10% heat-inactivated AB human serum, all purchased from PAA GmbH, and β2-mercaptoethanol from Gibco) for 2 h at 37°C. The blocking medium was removed and freshly isolated human PBMC (8×105 cells) were cultured with recombinant hnRNP-A2 protein (10 μg/mL), hnRNP-A2 peptides (10 μM), PPD of tuberculin (10 μg/mL), TT (10 μg/mL), or PHA (1/50), in a final volume of 200 μL complete RPMI medium. Control wells contained PBMC with medium alone. After 18- to 24-h incubation at 37°C, cells were removed, plates were washed with PBS/0.

Correlations with hospitalisations, deaths and renal failure will

Correlations with hospitalisations, deaths and renal failure will follow. Comparisons with other public practices,

with private renal practices, and by region and ethnic group will be interesting. 201 THE LUPUS NEPHRITIS AUSTRALIAN REGISTRY (LUNAR) R PHOON1, CHIR-99021 N ISBEL2, F BROWN3, P COATES4, K WYBURN5, R LANGHAM6, M LUTHERBORROW7, N KURSTJENS7, A IRISH8 1Westmead Hospital, Westmead, NSW; 2Princess Alexandra Hospital, Wooloongabba, QLD; 3Monash Medical Centre, Clayton, Victoria,4Royal Adelaide Hospital, Adelaide, SA; 5Royal Prince Alfred Hospital, Camperdown, NSW; 6St Vincent’s Hospital, Fitzroy, Victoria; 7Novartis Pharmaceuticals Australia, North Ryde, NSW; 8Royal Perth Hospital, Perth, WA, Australia Aim: To assess the safety, efficacy and outcomes of indigenous and non-indigenous patients treated for LN with Mycophenolate and other immunosuppressive agents within Australia. Background: Patients with Systemic lupus erythematosus (SLE) and kidney involvement, particularly WHO class III or IV lupus nephritis (LN), typically have poorer outcomes than those without. Until recently, the management of severe disease has involved corticosteroids and cyclophosphamide for both for induction and maintenance therapy. In 2012 mycophenolate sodium was approved in Australia for induction

and maintenance therapy in adult patients with WHO class III, IV or V LN. Methods: This is an ongoing multicentre, non-interventional study of patients click here treated for WHO class III, IV or V LN. Data is to be collected from approximately 200 patients taking mycophenolate sodium and other immunosuppressives over a 5 year period. Observational data capture includes laboratory measures of disease (serum creatinine and complement levels, full blood count, ESR, CRP, anti-dsDNA and urinary estimations of erythrocytes and proteinuria) and histopathology. Results: As of 31st March 2014 there is currently 81 patients recruited (41%

Caucasian, 8% Aboriginal/Torres Strait Islander, 30% Asian, 20% Other) with 85% of patients female and a mean age of 38 years. 46% Cytidine deaminase of patients are on a mycophenolate sodium regimen, 21% mycophenolate mofetil, 7% azathioprine 3% cyclophosphamide. Patients have a mean SLE disease duration of 9.9 years with a mean duration of LN of 6.15 years. Conclusions: LUNAR is the first study in Australia to examine outcomes in patients treated for WHO class III, IV or V LN with Mycophenolate and other immunosuppressive agents. 202 UTILISING EXOME SEQUENCING TO IDENTIFY NEPHRONOPHTHISIS MUTATIONS WITHIN AN AUSTRALIAN CLINICAL COHORT A MALLAWAARACHCHI1, A MALLETT2,3, A SAWYER4,5, H MCCARTHY4,5, J FLETCHER6, J CHAPMAN7, B BENNETTS8, G HO8, H JUEPPNER9, D HAHN4, S ALEXANDER4,5 1Department of Clinical Genetics, Westmead Hospital, New South Wales; 2Department of Renal Medicine, Royal Brisbane and Women’s Hospital, Queensland; 3CKD.