Treatment with Anti-Interleukin 23 Antibody Ameliorates Disease in Lupus-Prone Mice

Interleukin 23 receptor expressing IL-17 producing T cells have been shown to be important in the development of murine lupus. The usefulness of IL-23 inhibition in ameliorating lupus nephritis is unknown. We hypothesized that inhibition of IL-23 will ameliorate nephritis in lupus-prone mice. To this end, we treated MRL/lpr lupus-prone mice for 6 weeks with a rat anti-IL-23p19 antibody, which resulted in delaying the onset of nephritis without affecting the production of anti-dsDNA antibodies. The effect of the treatment was hampered by the production of murine anti-rat IgG antibodies. The amelioration of murine lupus by IL-23 inhibition strengthens the rationale for targeting IL-23 in patients with systemic lupus erythematosus

Μοριακή διερεύνηση ασθενών με νόσο Wilson

EΙΣΑΓΩΓΗ: Η νόσος Wilson (WD) είναι ένα κληρονομικό νόσημα που χαρακτηρίζεται από τη συσσώρευση χαλκού στο ήπαρ, τον εγκέφαλο και τους οφθαλμούς. Κληρονομείται με αυτοσωμικό υπολειπόμενο τρόπο και η συχνότητα εμφάνισης είναι 1/30000. ΣΚΟΠΟΣ: Σκοπός της εργασίας ήταν η ανάλυση ολόκληρης της κωδικοποιούσας αλληλουχίας του γονιδίου ATP7B για την ανίχνευση μεταλλάξεων που προκαλούν τη νόσο. Ο έλεγχος πραγματοποιήθηκε σε άτομα που παραπέμφθηκαν στο Εργαστήριο Ιατρικής Γενετικής του Εθνικού και Καποδιστριακού Πανεπιστημίου Αθηνών με υποψία νόσου Wilson. ΥΛΙΚΑ-ΜΕΘΟΔΟΛΟΓΙΑ: H ομάδα μελέτης αποτελείται από 30 ασθενείς με υποψία νόσου Wilson. O μοριακός έλεγχος του γονιδίου ATP7B πραγματοποιήθηκε με αλληλoύχηση κατά Sanger όλων των κωδικοποιουσών περιοχών και των παρακείμενων περιοχών στα όρια εσωνίων-εξωνίων. ΑΠΟΤΕΛΕΣΜΑΤΑ: Βρέθηκαν 14 διαφορετικές μεταλλάξεις ATP7B σε 16 ασθενείς με υποψία νόσου Wilson. Oι μεταλλάξεις που ανιχνεύθηκαν είναι οι ακόλουθες: c.3525delA (p.Gly1176Aspfs*16), c.3689T>C (p.Ile1230Thr), c.3207C>A (p.His1069Gln), c.2827G>A (p.Gly943Ser), c.1707+3insT, c.3400delC (p.Ala1135Glnfs*13), c.3688A>G (p.Ile1230Val), c.4396T>C (p.Ter1466Arg), c.2906G>A (p.Arg969Gln), c.3506T>C (p.Met1169Thr), c.2530delA (p.Val845Serfs*28), c.3295G>A (p.Gly1099Ser), c.1995G>A (p.Met665Ile) και η c.3284A>C (p.Gln1095Pro). ΣΥΜΠΕΡΑΣΜΑΤΑ: Οι μεταλλάξεις που έχουν ταυτοποιηθεί στο γονίδιο ATP7B αποτελούν την αιτία εκδήλωσης της νόσου Wilson. Η μελέτη οδήγησε στην ανίχνευση 2 νέων μεταλλάξεων (c.3525delA και c.3689Τ>C) και 12 γνωστών παθολογικών μεταλλάξεων. Σε 13 ασθενείς δεν ανιχνεύθηκαν μεταλλάξεις.INTRODUCTION: Wilson’s disease (WD) is a genetic disorder characterised by accumulation of copper in the liver, brain and eyes. It is inherited with autosomal recessive pattern. Its prevalence is estimated at 1/30000. PURPOSE: The purpose of this study was to sequence the ATP7B gene in order to detect mutations that cause Wilson disease that the patients were referred to the Medical Genetics laboratory of National and Kapodistrian University of Athens. MATERIALS-METHODS: The study group consists of 30 persons with possible Wilson disease. The coding portions and adjoining intronic sequences of the ATP7B gene were sequenced by Sanger sequencing. RESULTS: Sequencing of the ATP7B gene detected, 14 different mutations in 16 individuals with possible Wilson disease. The following mutations were detected: c.3525delA (p.Gly1176Aspfs*16), c.3689T>C (p.Ile1230Thr), c.3207C>A (p.His1069Gln), c.2827G>A (p.Gly943Ser), c.1707+3insT, c.3400delC (p.Ala1135Glnfs*13), c.3688A>G (p.Ile1230Val), c.4396T>C (p.Ter1466Arg), c.2906G>A (p.Arg969Gln), c.3506T>C (p.Met1169Thr), c.2530delA (p.Val845Serfs*28), c.3295G>A (p.Gly1099Ser), c.1995G>A (p.Met665Ile) and c.3284A>C (p.Gln1095Pro). CONCLUSIONS: Mutations in ATP7B gene cause Wilson disease. The study lead to the detection of 2 new mutations (c.3525delA and c.3689T>C) and 12 previously characterized mutations. No mutations were detected in 13 individuals

Novel Treatments in Lupus

Purpose of Review: The standard treatment options for systemic lupus erythematosus (SLE) are focused on non-specific immunosuppression. Over the past few years, scientific studies and ongoing clinical trials have shifted the paradigm with rapid advances in developing biologics and small molecules. A number of monoclonal antibodies and small molecule inhibitors have been developed to target specific pathways involved in SLE. Many of these novel therapeutic agents are already being tested in clinical trials and they may 1 day reshape the landscape of SLE treatment. Herein we review potential future therapeutic options for SLE

SLE serum deposits C4d on red blood cells, decreases red blood cell membrane deformability, and promotes nitric oxide production

Objective Systemic lupus erythematosus (SLE) is characterized by intravascular activation of the complement system and deposition of complement fragments (C3 and C4) on plasma membranes of circulating cells, including red blood cells (RBC). The aim of this study was to address whether this process affects the biophysical properties of RBC. Methods Serum and red blood cells were isolated from patients with SLE, and healthy controls. RBC from healthy O Rh negative individuals were incubated with SLE or control serum. We used flow cytometry to assess complement fragment deposition on RBC. RBC membrane deformability was measured using 2D microchannel arrays. Protein phosphorylation levels were quantified by western blot. Results Incubation of healthy donor RBC with sera from patients with SLE but not control sera led to deposition of C4 fragments on the RBC. Complement decorated RBC exhibited significant decrease in both membrane deformability and flickering. Sera from SLE patients triggered a transitory Ca++ influx in RBC that was associated with decreased phosphorylation of ?-spectrin, and increased phosphorylation of band 3, two key proteins of RBC cytoskeleton. Finally, SLE but not control sera led to the production of nitric oxide (NO) by RBC. Conclusion Our data suggest that complement activation in patients with SLE leads to calcium dependent cytosketeletal changes in RBC that render them less deformable, likely impairing their flow through capillaries. This phenomenon may negatively impact the delivery of oxygen to the tissues

Spleen Tyrosine Kinase (Syk) Regulates Systemic Lupus Erythematosus (SLE) T Cell Signaling

Engagement of the CD3/T cell receptor complex in systemic lupus erythematosus (SLE) T cells involves Syk rather than the zeta-associated protein. Because Syk is being considered as a therapeutic target we asked whether Syk is central to the multiple aberrantly modulated molecules in SLE T cells. Using a gene expression array, we demonstrate that forced expression of Syk in normal T cells reproduces most of the aberrantly expressed molecules whereas silencing of Syk in SLE T cells normalizes the expression of most abnormally expressed molecules. Protein along with gene expression modulation for select molecules was confirmed. Specifically, levels of cytokine IL-21, cell surface receptor CD44, and intracellular molecules PP2A and OAS2 increased following Syk overexpression in normal T cells and decreased after Syk silencing in SLE T cells. Our results demonstrate that levels of Syk affect the expression of a number of enzymes, cytokines and receptors that play a key role in the development of disease pathogenesis in SLE and provide support for therapeutic targeting in SLE patients

Identification of Orch3, a Locus Controlling Dominant Resistance to Autoimmune Orchitis, as Kinesin Family Member 1C

Experimental autoimmune orchitis (EAO), the principal model of non-infectious testicular inflammatory disease, can be induced in susceptible mouse strains by immunization with autologous testicular homogenate and appropriate adjuvants. As previously established, the genome of DBA/2J mice encodes genes that are capable of conferring dominant resistance to EAO, while the genome of BALB/cByJ mice does not and they are therefore susceptible to EAO. In a genome scan, we previously identified Orch3 as the major quantitative trait locus controlling dominant resistance to EAO and mapped it to chromosome 11. Here, by utilizing a forward genetic approach, we identified kinesin family member 1C (Kif1c) as a positional candidate for Orch3 and, using a transgenic approach, demonstrated that Kif1c is Orch3. Mechanistically, we showed that the resistant Kif1c$^{D2}$ allele leads to a reduced antigen-specific T cell proliferative response as a consequence of decreased MHC class II expression by antigen presenting cells, and that the L$^{578}$→P$^{578}$ and S$^{1027}$→P$^{1027}$ polymorphisms distinguishing the BALB/cByJ and DBA/2J alleles, respectively, can play a role in transcriptional regulation. These findings may provide mechanistic insight into how polymorphism in other kinesins such as KIF21B and KIF5A influence susceptibility and resistance to human autoimmune diseases

ICER is requisite for Th17 differentiation.

Inducible cAMP early repressor (ICER) has been described as a transcriptional repressor isoform of the cAMP response element modulator (CREM). Here we report that ICER is predominantly expressed in Th17 cells through the IL-6-STAT3 pathway and binds to the Il17a promoter, where it facilitates the accumulation of the canonical enhancer RORγt. In vitro differentiation from naive ICER/CREM-deficient CD4(+) T cells to Th17 cells is impaired but can be rescued by forced overexpression of ICER. Consistent with a role of Th17 cells in autoimmune and inflammatory diseases, ICER/CREM-deficient B6.lpr mice are protected from developing autoimmunity. Similarly, both anti-glomerular basement membrane-induced glomerulonephritis and experimental encephalomyelitis are attenuated in ICER/CREM-deficient mice compared with their ICER/CREM-sufficient littermates. Importantly, we find ICER overexpressed in CD4(+) T cells from patients with systemic lupus erythematosus. Collectively, our findings identify a unique role for ICER, which affects both organ-specific and systemic autoimmunity in a Th17-dependent manner

The CD38/NAD/SIRTUIN1/EZH2 Axis Mitigates Cytotoxic CD8 T Cell Function and Identifies Patients with SLE Prone to Infections

Summary: Patients with systemic lupus erythematosus (SLE) suffer frequent infections that account for significant morbidity and mortality. T cell cytotoxic responses are decreased in patients with SLE, yet the responsible molecular events are largely unknown. We find an expanded CD8CD38high T cell subset in a subgroup of patients with increased rates of infections. CD8CD38high T cells from healthy subjects and patients with SLE display decreased cytotoxic capacity, degranulation, and expression of granzymes A and B and perforin. The key cytotoxicity-related transcription factors T-bet, RUNX3, and EOMES are decreased in CD8CD38high T cells. CD38 leads to increased acetylated EZH2 through inhibition of the deacetylase Sirtuin1. Acetylated EZH2 represses RUNX3 expression, whereas inhibition of EZH2 restores CD8 T cell cytotoxic responses. We propose that high levels of CD38 lead to decreased CD8 T cell-mediated cytotoxicity and increased propensity to infections in patients with SLE, a process that can be reversed pharmacologically. : Katsuyama et al. find that an expanded CD8CD38high T cell population in SLE patients is linked to infections. CD8CD38high T cells display decreased cytotoxic capacity by suppressing the expression of related molecules through an NAD+/Sirtuin1/EZH2 pathway. EZH2 inhibitors increase cytotoxicity offering a means to mitigate infection rates in SLE. Keywords: systemic lupus erythematosus, patients, CD8 T cell, CD38, cytotoxicity, infection, nicotinamide adenine dinucleotide, Sirtuin1, EZH

Diet Influences Expression of Autoimmune‐Associated Genes and Disease Severity by Epigenetic Mechanisms in a Transgenic Mouse Model of Lupus

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/98819/1/art37967.pd

NFATc1 supports imiquimod-induced skin inflammation by suppressing IL-10 synthesis in B cells

Epicutaneous application of Aldara cream containing the TLR7 agonist imiquimod (IMQ) to mice induces skin inflammation that exhibits many aspects of psoriasis, an inflammatory human skin disease. Here we show that mice depleted of B cells or bearing interleukin (IL)-10-deficient B cells show a fulminant inflammation upon IMQ exposure, whereas ablation of NFATc1 in B cells results in a suppression of Aldara-induced inflammation. In vitro, IMQ induces the proliferation and IL-10 expression by B cells that is blocked by BCR signals inducing NFATc1. By binding to HDAC1, a transcriptional repressor, and to an intronic site of the Il10 gene, NFATc1 suppresses IL-10 expression that dampens the production of tumour necrosis factor-α and IL-17 by T cells. These data indicate a close link between NFATc1 and IL-10 expression in B cells and suggest NFATc1 and, in particular, its inducible short isoform, NFATc1/αA, as a potential target to treat human psoriasis