Genetic Ablation of Nrf2 Exacerbates Neuroinflammation in Ocular Autoimmunity.

Experimental autoimmune uveoretinitis (EAU) is an animal model of non-infectious uveitis and is developed by immunization with retinal antigen, interphotoreceptor retinoid-binding protein (IRBP). Nuclear factor erythroid 2- (NF-E2-) related factor 2 (Nrf2) is responsible for regulating antioxidant and inflammatory responses. In this study, we investigated the role of Nrf2 on the development of EAU. Clinical and pathological examination demonstrated that retinal inflammation was exacerbated in Nrf2 knockout (Nrf2 KO) mice compared to wild type (WT) mice, and the expression of inflammatory cytokines (IFN-γ, IL-6, and IL-17) in the retina was significantly elevated in Nrf2 KO mice. GFAP positive cells (astrocytes) and Iba-1 positive cells (microglia cells) in the retina were more numerous in Nrf2 KO mice compared to WT mice. Furthermore, we examined the suppressive effect of the Nrf2 activator CDDO-Im (2-cyano-3,12 dioxooleana-1,9 dien-28-oyl imidazoline) on the development of EAU. The treatment with CDDO-Im significantly reduced the clinical and pathological score of EAU compared to those of vehicle-treated mice. These findings suggest that Nrf2 plays a regulatory role in the pathogenesis of autoimmune uveoretinitis and the activation of the Nrf2 system may have therapeutic potential for protecting vision from autoimmune neuroinflammation.

Mixed-diphosphine-protected chiral undecagold clusters Au11(S,S-DIOP)4(rac-/R-/S-BINAP): effect of the handedness of BINAP on their chiroptical responses.

In this article, we report a preference of homochiral-type ligation of BINAP that produces SS-type ligand assembly onto the Au11 clusters protected by diphosphine S,S-DIOP. The Au11 clusters synthesized and isolated are Au11(S,S-DIOP)4(rac-/R-/S-BINAP), and their optical/chiroptical responses are characterized. Absorption spectra of these Au11 clusters are almost identical to each other, but their CD profiles are dependent on the handedness of BINAP. In Au11(S,S-DIOP)4(rac-BINAP), the yield of S-BINAP or R-BINAP coordination is roughly comparable, but we found a small but distinctive preference in the S-BINAP ligation; that is, homochiral-type (SS-type) ligand assembly formation. Quantum chemical calculations for simplified model clusters suggest equal contributions of S- and R-form BINAP coordination. The experimentally-observed preference of homochiral-type ligation can then be due to that of the whole ligand structures and assemblies involving interligand interactions. Chiral sorting and amplification processes through the assembly control of homochirality or heterochirality are of primary importance for the development of enantioselective reactions, so we anticipate this finding will contribute to further understanding of such processes based on various metal clusters with chiral ligands.

Chirality in Au9 clusters protected by chiral/achiral mixed bidentate phosphine ligands: influence of the metal core and ligand array.

In this article, the chiroptical responses of Au9 clusters protected by chiral/achiral mixed bidentate phosphine ligands are reported. The mixed phosphine we use is (S)-BINAP/Xantphos in the molar ratio of 1/0 (= pure (S)-BINAP), 3/1, 1/1, or 0/1 (= pure Xantphos), where BINAP and Xantphos represent 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, respectively. Electronic absorption spectra of the clusters are similar between the samples with different molar diphosphine ratios, but the chiroptical activity or g-factor decreases nonlinearly with an increase in the fraction of Xantphos. Quantum chemical calculations and geometrical quantifications based on the Hausdorff chirality measure (HCM) for model Au9 cluster species suggest that (i) two types of metal core structures with pseudo-P- and M-chirality are found, and their appropriate contributions would cancel out the chiroptical response in the low-energy region; (ii) the origin of optical activity in pure (S)-BINAP-protected Au9 clusters can mainly be attributed to the metal core chirality, whereas that of other mixed-ligand protected clusters would be due to the chiral ligand arrangement. This work demonstrates that the modulation of chiroptical activity in Au9 clusters by chiral/achiral mixed-diphosphine ligation is controlled by the difference in the degree of chirality existing in the cluster core and/or the ligand array.

Dehydroxymethylepoxyquinomicin, a novel nuclear factor-κB inhibitor, reduces chemokines and adhesion molecule expression induced by IL-1ß in human corneal fibroblasts.

PURPOSE: Dehydroxymethylepoxyquinomicin (DHMEQ) is derived from the antibiotic, epoxyquinomicin C, and is a novel low molecular weight nuclear factor-κB (NF-κB) inhibitor. We investigated the effects of DHMEQ on the expression of chemokines and the intercellular adhesion molecule (ICAM)-1 induced by proinflammatory cytokines in cultures of the human corneal fibroblasts (HCFs). METHODS: The cytotoxicity of DHMEQ on cultured HCFs was evaluated by cell proliferation assays. Cultures were exposed to interleukin (IL)-1ß, and the production of IL-8 and monocyte chemoattractant protein (MCP)-1 was assessed by enzyme-linked immunosorbent assay. The degree of expression of ICAM-1 was measured by flow cytometry. The translocation of NF-κB p65 into the nucleus of HCFs was assessed by immunocytochemistry. RESULTS: DHMEQ was not toxic to cultured HCFs at doses up to 10 µg/ml. DHMEQ significantly suppressed the production of both IL-8 and MCP-1 in IL-1ß-stimulated HCFs. In addition, DHMEQ down-regulated ICAM-1 expression in IL-1ß-stimulated HCFs in a dose-dependent manner. DHMEQ inhibited the IL-1ß-induced nuclear accumulation of p65, a component of NF-κB, in HCFs. CONCLUSIONS: The suppression of inflammatory chemokines IL-8 and MCP-1 and inhibition of the expression of ICAM-1 in cultured HCFs by DHMEQ indicates that DHMEQ may have a therapeutic potential for treating ICAM-1 and chemokine-mediated corneal inflammatory disorders.

Retinoic acid receptor stimulation ameliorates experimental autoimmune optic neuritis.

BACKGROUND: To determine whether all-trans retinoic acid or a synthetic retinoic acid receptor-α/ß-specific agonist, Am80, can reduce the degree of experimental autoimmune optic neuritis in mice with experimental autoimmune encephalomyelitis. METHODS: Optic neuritis was induced in C57BL/6 mice by immunizing them with myelin oligodendrocyte glycoprotein35-55 . All-trans retinoic acid (350 µg/mouse/time point) or Am80 (5 mg/kg/time point) was administered every other day from day 0 to day 20. The degree of experimental autoimmune encephalomyelitis was scored and histopathological analysis of the optic neuritis was performed on day 22 after the immunization. In vivo-primed draining lymph node cells obtained from vehicle-treated or all-trans retinoic acid-treated mice were stimulated with myelin oligodendrocyte glycoprotein35-55 , and the culture supernatant was collected for assays of interferon-γ and interleukin-17. RESULTS: All-trans retinoic acid treatment significantly reduced the clinical score of experimental autoimmune encephalomyelitis and the severity of the optic neuritis by histopathological analysis. The production of interferon-γ and interleukin-17 was significantly reduced in all-trans retinoic acid-treated mice compared with vehicle-treated mice. Am80 treatment also significantly decreased the severity of the optic neuritis in mice with experimental autoimmune encephalomyelitis. CONCLUSIONS: These findings demonstrate that all-trans retinoic acid and Am80 treatment were able to reduce the severity of optic neuritis in mice with experimental autoimmune encephalomyelitis. Activation of retinoic acid receptor-α/ß may be a molecular target for the treatment of autoimmune optic neuritis induced by Th1 or Th17-dominated immune responses.

Structural basis for docking of peroxisomal membrane protein carrier Pex19p onto its receptor Pex3p.

Peroxisomes require peroxin (Pex) proteins for their biogenesis. The interaction between Pex3p, which resides on the peroxisomal membrane, and Pex19p, which resides in the cytosol, is crucial for peroxisome formation and the post-translational targeting of peroxisomal membrane proteins (PMPs). It is not known how Pex3p promotes the specific interaction with Pex19p for the purpose of PMP translocation. Here, we present the three-dimensional structure of the complex between a cytosolic domain of Pex3p and the binding-region peptide of Pex19p. The overall shape of Pex3p is a prolate spheroid with a novel fold, the 'twisted six-helix bundle.' The Pex19p-binding site is at an apex of the Pex3p spheroid. A 16-residue region of the Pex19p peptide forms an α-helix and makes a contact with Pex3p; this helix is disordered in the unbound state. The Pex19p peptide contains a characteristic motif, consisting of the leucine triad (Leu18, Leu21, Leu22), and Phe29, which are critical for the Pex3p binding and peroxisome biogenesis.

Spatial reorganization of Saccharomyces cerevisiae enolase to alter carbon metabolism under hypoxia.

Hypoxia has critical effects on the physiology of organisms. In the yeast Saccharomyces cerevisiae, glycolytic enzymes, including enolase (Eno2p), formed cellular foci under hypoxia. Here, we investigated the regulation and biological functions of these foci. Focus formation by Eno2p was inhibited temperature independently by the addition of cycloheximide or rapamycin or by the single substitution of alanine for the Val22 residue. Using mitochondrial inhibitors and an antioxidant, mitochondrial reactive oxygen species (ROS) production was shown to participate in focus formation. Focus formation was also inhibited temperature dependently by an SNF1 knockout mutation. Interestingly, the foci were observed in the cell even after reoxygenation. The metabolic turnover analysis revealed that [U-(13)C]glucose conversion to pyruvate and oxaloacetate was accelerated in focus-forming cells. These results suggest that under hypoxia, S. cerevisiae cells sense mitochondrial ROS and, by the involvement of SNF1/AMPK, spatially reorganize metabolic enzymes in the cytosol via de novo protein synthesis, which subsequently increases carbon metabolism. The mechanism may be important for yeast cells under hypoxia, to quickly provide both energy and substrates for the biosynthesis of lipids and proteins independently of the tricarboxylic acid (TCA) cycle and also to fit changing environments.

Development of formulation device for periodontal disease.

In addition to providing standard surgical treatment that removes the plaque and infected tissues, medications that can regenerate periodontal tissue are also required in the treatment of periodontal disease. As a form of regenerative medication, various growth factors are expected to be used while treating periodontal disease. A protein-like growth factor is often developed as a lyophilized product with dissolution liquid, considering its instability in the solution state. We have clarified that the formulation for periodontal disease needs to be viscous. When the lyophilized product was dissolved using a sticky solution, various problems were encountered, difficulty in dissolving and air bubbles, for example, and some efforts were needed to prepare the formulation. In this research, to identify the problem of preparing a viscous formulation, a lyophilized product (placebo) and sticky liquid were prepared by using vial and ampoule as the conventional containers. Based on these problems, a prototype administration device was developed, and its functionality was confirmed. As a result, it was suggested that the device with a useful mixing system that could shorten the preparation time was developed.

Effect of In Vivo Expansion of Regulatory T Cells with IL-2/anti-IL-2 Antibody Complex Plus Rapamycin on Experimental Autoimmune Uveoretinitis.

Purpose: To determine the effect of injection of IL-2/anti-IL-2 antibody (IL-2 complex) together with rapamycin on the development of experimental autoimmune uveoretinitis (EAU).Methods: C57BL/6J mice were immunized with human interphotoreceptor retinoid-binding protein peptide. The immunized mice were injected intraperitoneally with PBS, IL-2 complex, rapamycin, or IL-2 complex/rapamycin on days 1, 2, 3, and 4 (induction phase) or days 10, 11, 12, and 13 (effector phase) after immunization.Results: Expansion of CD4+Foxp3+ regulatory T cells in draining lymph nodes was observed in IL-2 complex and IL-2 complex/rapamycin-treated mice. Although injection of IL-2 complex alone was not capable of decreasing the clinical score of EAU, injection of IL-2 complex/rapamycin significantly delayed the onset of EAU. In contrast, the treatment with IL-2 complex alone or IL-2 complex/rapamycin during effector phase failed to suppress EAU.Conclusions: These findings suggest the potential limitations of IL-2 complex or IL-2 complex/rapamycin during EAU.

Evaluation of additives required for periodontal disease formulation using basic fibroblast growth factor.

To design a suitable periodontal disease formulation using basic fibroblast growth factor (bFGF), legally available thickeners were evaluated focusing on their viscosity, extrusive force from a syringe, flow property and inertness to bFGF. Thirteen candidate thickeners showed appropriate viscosity (about 1×104 mPa·s), and further evaluations were conducted on them. Flow property was evaluated by the tilting test tube method. As a result, most thickener solutions with the optimum viscosity showed appropriate flow time (about 100 s) and the flow time did not depend on thickener concentration, whereas the extrusive force from a syringe depended on thickener concentration despite the thickener type and grade. Thickener solutions of 2-3% showed ideal result (10-20 N) and thickener solutions prepared outside of the concentration range (2-3%) were found to show unsuitable extrusive force. Consequently, to obtain required properties for a dental drug formulation, thickener solutions needed to show adequate viscosity (about 1×104 mPa·s) at 2-3% thickener concentration. In addition, several types of cellulose derivatives showed inertness to the bFGF because of their structure, without strong ionic dissociable groups, and neutral pH. Overall, the present work demonstrates that some water-soluble cellulose derivatives, such as hydroxypropylcellulose (HPC) and hydroxyethylcellulose (HEC), were suggested to have required properties for a dental drug formulation including bFGF.

Anti­inflammatory effects of the NF­κB inhibitor dehydroxymethylepoxyquinomicin on ARPE­19 cells.

The retinal pigment epithelium (RPE) is a polarized, monolayer of pigmented cells that forms the outer retinal layer. A key function of the RPE is to maintain the integrity of the photoreceptors mainly via phagocytosis and recycling of the digested photoreceptor outer segments. Moreover, RPE cells are a major source of inflammatory cytokines and chemokines, which play important roles in the activation of other immune cells under inflammatory conditions in the posterior segment of the eye. Dehydroxymethylepoxyquinomicin (DHMEQ) is a NF­κB inhibitor and its structure is related to that of epoxyquinomicin C, which is an antibiotic. The present study evaluated the anti­inflammatory effects of DHMEQ on a human retinal pigment epithelial cell line (ARPE­19). It was revealed that high concentrations of DHMEQ (100 µg/ml) induced apoptosis and necrosis of tumor necrosis factor (TNF)­α­stimulated ARPE­19 cells. Furthermore, the percentage of intercellular adhesion molecule 1 (ICAM­1)­positive TNF­α­stimulated cells was significantly reduced in the presence of DHMEQ (10 µg/ml), as determined by flow cytometry. It was also demonstrated that DHMEQ exposure significantly decreased the levels of interleukin (IL)­8 and monocyte chemoattractant protein­1 (MCP­1) in the supernatant of cultured ARPE­19 cells as determined by ELISA. Moreover, the protein expression levels of IL­8 and MCP­1 were significantly reduced in ARPE­19 cells exposed to DHMEQ compared with cells exposed to dexamethasone. PCR array analysis revealed that DHMEQ reduced the expression levels of MCP­1, ICAM­1, IL­6, Toll­like receptor (TLR)2, TLR3 and TLR4. Therefore, the present results indicated that DHMEQ has anti­inflammatory effects on TNF­α­stimulated ARPE­19 cells. Thus, DHMEQ may have therapeutic potential for TNF­α­mediated inflammatory disorders of the eye.

Massive and Reproducible Production of Liver Buds Entirely from Human Pluripotent Stem Cells.

Organoid technology provides a revolutionary paradigm toward therapy but has yet to be applied in humans, mainly because of reproducibility and scalability challenges. Here, we overcome these limitations by evolving a scalable organ bud production platform entirely from human induced pluripotent stem cells (iPSC). By conducting massive "reverse" screen experiments, we identified three progenitor populations that can effectively generate liver buds in a highly reproducible manner: hepatic endoderm, endothelium, and septum mesenchyme. Furthermore, we achieved human scalability by developing an omni-well-array culture platform for mass producing homogeneous and miniaturized liver buds on a clinically relevant large scale (>108). Vascularized and functional liver tissues generated entirely from iPSCs significantly improved subsequent hepatic functionalization potentiated by stage-matched developmental progenitor interactions, enabling functional rescue against acute liver failure via transplantation. Overall, our study provides a stringent manufacturing platform for multicellular organoid supply, thus facilitating clinical and pharmaceutical applications especially for the treatment of liver diseases through multi-industrial collaborations.

MicroRNAs in retina during development of experimental autoimmune uveoretinitis in rats.

PURPOSE: To determine the changes in the expression profiles of microRNAs (miRNAs) in retinas during the development of experimental autoimmune uveoretinitis (EAU) in rats. METHODS: The levels of interleukin-1ß (IL-1ß) and monocyte chemoattractant protein-1 (MCP-1) were measured in aqueous humour samples and supernatants of homogenised posterior eye cups obtained from Lewis rats immunised with interphotoreceptor retinoid binding protein peptide (R14) and complete Freund's adjuvant. Microarray analysis was performed to determine the miRNA profiles in the retina of eyes with EAU on days 0 (baseline), 7, 14 and 21 after immunisation. RESULTS: The levels of IL-1ß and MCP-1 in the aqueous humour and the supernatants of posterior eye cups were significantly elevated in eyes with EAU, and the levels corresponded with the stage of the EAU. On day 14 after immunisation, the expressions of nine miRNAs (miRNA-223, 142-5p, 142-3p, 21, 146a, 146b, 1949, 1188-3p and 193) were significantly elevated, and the expressions of four miRNAs (miRNA-181a, 183*, 124* and 331) were downregulated relative to the baseline. Quantitative PCR analyses confirmed the elevation of miRNA-223 and miRNA-146 and the downregulation of miRNA-181a in retinas with EAU on day 14 after immunisation. In situ hybridisation confirmed increased expression of miR-223 and miR-146 in retinas with EAU. CONCLUSIONS: Several miRNAs were significantly increased or decreased in retinas during the course of EAU. The expression of miR-223 and miR-146a corresponded with the clinical score of the EAU and elevation of IL-1ß/MCP-1 in the eye with EAU. Further studies are required to clarify the role of miRNA in eyes with autoimmune uveoretinitis.

Pulsed holmium:yttrium-aluminum-garnet laser-induced liquid jet as a novel dissection device in neuroendoscopic surgery.

OBJECT: A pressure-driven continuous jet of water has been reported to be a feasible tool for neuroendoscopic dissection owing to its superiority at selective tissue dissection in the absence of thermal effects. With respect to a safe, accurate dissection, however, continuous water flow may not be suitable for intraventricular use. The authors performed experiments aimed at solving problems associated with continuous flow by using a pulsed holmium:yttrium-aluminum-garnet (Ho:YAG) laser-induced liquid jet (LILJ). They present this candidate neuroendoscopic LILJ dissection system, having examined its mechanical characteristics and evaluated its controllability both in a tissue phantom and in a rabbit cadaveric ventricle wall. METHODS: The LILJ generator was incorporated into the tip of a No. 4 French catheter so that the LILJ could be delivered via a neuroendoscope. Briefly, the LILJ was generated by irradiating an internally supplied column of physiological saline with a pulsed Ho:YAG laser (pulse duration time 350 microsec; laser energy 250-700 mJ/pulse) within a No. 4 French catheter (internal diameter 1 mm) and ejecting it from a metal nozzle (internal diameter 100 microm). The Ho:YAG laser energy pulses were conveyed by an optical fiber (core diameter 400 microm) at 3 Hz, whereas physiological saline (4 degrees C) was supplied at a rate of 40 ml/hour. The mechanical characteristics of the pulsed LILJ were investigated using high-speed photography and pressure measurements; thermal effects and controllability were analyzed using an artificial tissue model (10% gelatin of 1 mm thickness). Finally, the ventricle wall of a rabbit cadaver was dissected using the LILJ. Jet pressure increased in accordance with laser energy from 0.1 to 2 bar; this translated into a penetration depth of 0.08 to 0.9 mm per shot in the ventricle wall of the rabbit cadaver. The gelatin phantom could be cut into the desired shape without significant thermal effects and in the intended manner, with a good surgical view. CONCLUSIONS: The present results show that the pulsed LILJ has the potential to become a safe and reliable dissecting method for endoscopic procedures.

Hypoglossal neurinoma presenting with intratumoral hemorrhage.

Focal or microscopic hemorrhage in a neurinoma is common, but tumor origin from the hypoglossal nerve and extensive symptomatic intratumoral hemorrhage are both rare. A 59-year-old male presented with severe neck pain, nausea and vomiting of 1-day duration, accompanied by right hypoglossal nerve palsy. Neuroimaging disclosed a tumor located in the right cerebellomedullary fissure and containing a hematoma. The right hypoglossal canal was slightly dilated. The intracranial tumor was resected via a suboccipital approach. Histological examination demonstrated spindle-shaped tumor cells with nuclear palisading and also relative hypervascularity with hyaline degeneration of the vessels. Extensive hemorrhage was present, as was necrosis. Thickening and hyalinization of arterial walls, a common occurrence in neurinomas, may have contributed to symptomatic intratumoral hemorrhage.

[Antimicrobial susceptibility and prevalence of beta-lactamase producing clinical isolates in southern Kyushu. The results of collaborative study from 1999 to 2000].

The positivity of beta-lactamase and antimicrobial susceptibility were determined in a total of 1,358 clinical isolates at 15 hospitals and clinics in four prefectures in southern Kyushu (Okinawa, Miyazaki, Kagoshima and Kumamoto) during the period from December 1999 to February 2000. The isolates collected comprised of 176 strains of S. aureus, 203 of H. influenzae, 102 of M. catarrhalis, 206 of E. coli, 153 of K. pneumoniae, 99 of E. cloacae, 95 of S. marcescens, 201 of P. aeruginosa, 79 of E. faecalis, and 44 of E. faecium. The frequency of CPDX resistance among E. coli in particular varied geographically, and was found to be higher in Kumamoto and Kagoshima. The strains of K. pneumoniae and E. cloacae resistant to common antimicrobial agents were particularly found in Kagoshima, and one strain of IPM-resistant E. cloacae was isolated in Miyazaki. Also, the geographical difference in the frequency of LVFX resistance among the isolates of E. cloacae was noted, the results indicating the higher prevalence in Okinawa and Kagoshima. Resistant isolates of P. aeruginosa were less common in Kagoshima, and four isolates of P. aeruginosa from Miyazaki were found to be resistant to CAZ and IPM. None of the isolates of S. aureus and Enterococcus spp. was resistant to VCM or TEIC at all. The isolates of E. faecalis resistant at high-level GM (500 micrograms/ml) and SM (1,000 micrograms/ml) were found in 27.8% and 22.8%, and those of E. faecium were 6.8% and 38.6%, respectively. Overall, the ratio of MRSA among S. aureus was 67.6%, and three isolates were resistant to ABK with no less than 8 micrograms/ml of MIC. The frequency of BLNAR (beta-lactamase-negative, ampicillin resistant) among H. influenzae isolated in Okinawa was markedly higher (isolation ratio, 37.9%) when compared with other prefectures, and the isolates of BLPACR (beta-lactamase-positive, AMPC/CVA resistant) were found only in Okinawa with a ratio of 41.6%. A total of 18 strains of ESBL defined by the NCCLS criteria (M100-S11) were isolated, eight strains of K. pneumoniae and 10 strains of E. coli. Of 18 isolates of ESBL, 13 were from Kagoshima and the remaining five were from Kumamoto.

[Effect of concomitant use of dental drug on the properties of recombinant human basic fibroblast growth factor formulation for periodontal disease].

We have discussed the essential property for periodontal disease medication using protein, such as recombinant human basic fibroblast growth factor (rhbFGF). In our previous study, the criteria of thickener for the medication, viscosity, flowability etc., were set. The aim of this study was to evaluate the physical and chemical effect of concomitant use of general dental drug or device on thickener properties for the clinical use of viscous rhbFGF formulation. Viscous formulation was prepared with six cellulose derivatives, two types hydroxy propyl cellulose (HPC), three types hydroxy ethyl cellulose (HEC) and methyl cellulose (MC). Antibiotic ointment, local anesthetic, bone graft substitute, agent for gargle and mouthwashes, were chosen as general dental drug and device. These drugs and device were mixed with the viscous formulations and the change of viscosity and flowability, the remaining ratio of rhbFGF were evaluated. When the various thickener solutions were mixed with the liquid drugs, viscosity and flowability did not changed much. However, in the case of MC solution, viscous property declined greatly when MC solution was mixed with cationic surfactant for gargle. The flowabilities of thickener solutions were declined with insoluble bone graft. The stabilities of rhbFGF in thickener solutions were no problem for 24 hours even in the case of mixing with dental drug or device. Our findings suggested that the viscous rhbFGF formulations prepared in this research were not substantially affected by the concomitant use of dental drug or device, especially the formulation with HPC or HEC was useful.

Oral administration of retinoic acid receptor-alpha/beta-specific ligand Am80 suppresses experimental autoimmune uveoretinitis.

PURPOSE: To determine whether synthetic retinoic acid receptor (RAR)-α/ß-specific agonist Am80 reduces inflammation in experimental autoimmune uveoretinitis (EAU). METHODS: Naive CD4(+) T cells were activated with anti-CD3, anti-CD28, and transforming growth factor (TGF)-ß, in the presence or absence of Am80. Intracellular expression of forkhead box p3 (Foxp3) and interleukin (IL)-17 in the activated CD4(+) T cells was assessed by flow cytometry. For induction of EAU, C57BL/6 mice were immunized with human interphotoreceptor retinoid binding protein (IRBP) peptide 1 to 20 (IRBP(1-20)). Am80 was administered orally every other day (3 mg/kg/time point) from day 0 to day 21. In vivo primed draining lymph node cells from vehicle-treated or Am80-treated mice were stimulated with IRBP(1-20), and culture supernatant was harvested for assay of interferon (IFN)-γ, IL-6, IL-10, and IL-17. The expression of Foxp3 and IL-6 receptor α in CD4(+) T cells of draining lymph node cells was assessed by a flow cytometer. RESULTS: Am80 synergized with TGF-ß to induce Foxp3(+) T regulatory cells (Treg) and reciprocally inhibited development of IL-17-producing T helper cells (Th17) induced by TGF-ß and IL-6. Am80 treatment reduced the severity of EAU clinically, and IFN-γ and IL-17 production was significantly reduced in Am80-treated mice. In addition, the expression of IL-6 receptor α on CD4(+) T cells was downregulated in Am80-treated mice. CONCLUSIONS: These findings demonstrate that Am80 treatment ameliorates severity of EAU and reduces the Th1/Th17 responses. The synthetic retinoid Am80 appears to be a promising agent for preventing autoimmune uveoretinal inflammation.

Anti-inflammatory effect of retinoic acid on experimental autoimmune uveoretinitis.

AIMS: To determine whether an active metabolite of vitamin A, all-trans retinoic acid (ATRA), reduces inflammation in experimental autoimmune uveoretinitis (EAU). METHODS: Naive CD4(+) T cells were activated with anti-CD3, anti-CD28 and transforming growth factor (TGF)-beta, in the presence or absence of ATRA. Intracellular expression of transcription factor forkhead box P3 (Foxp3) and interleukin (IL)-17 in the activated CD4(+) T cells was assessed by flow cytometry. C57BL/6 mice were immunised with human interphotoreceptor retinoid binding protein peptide 1-20 (IRBP(1-20)). ATRA was administered intraperitoneally every other day (0.2 mg/mouse per day) from day 0 to day 21. In vivo-primed draining lymph node cells from vehicle-treated or ATRA-treated mice were stimulated with IRBP(1-20) and the culture supernatant fraction was harvested for assay of interferon (IFN)-gamma and IL-17 by ELISA. RESULTS: ATRA synergised with TGF-beta to induce Foxp3(+) T regulatory cells (Treg) and reciprocally inhibited development of IL-17-producing T helper cells (Th17) induced by TGF-beta and IL-6. ATRA treatment reduced the severity of EAU clinically, and IFN-gamma and IL-17 production were significantly reduced in ATRA-treated mice. CONCLUSION: These findings demonstrate that ATRA treatment ameliorates severity of EAU and reduces the Th1/Th17 responses. ATRA may represent a new therapeutic modality for human refractory uveitis.