The E3 ligase synoviolin controls body weight and mitochondrial biogenesis through negative regulation of PGC-1ß.

Obesity is a major global public health problem, and understanding its pathogenesis is critical for identifying a cure. In this study, a gene knockout strategy was used in post-neonatal mice to delete synoviolin (Syvn)1/Hrd1/Der3, an ER-resident E3 ubiquitin ligase with known roles in homeostasis maintenance. Syvn1 deficiency resulted in weight loss and lower accumulation of white adipose tissue in otherwise wild-type animals as well as in genetically obese (ob/ob and db/db) and adipose tissue-specific knockout mice as compared to control animals. SYVN1 interacted with and ubiquitinated the thermogenic coactivator peroxisome proliferator-activated receptor coactivator (PGC)-1ß, and Syvn1 mutants showed upregulation of PGC-1ß target genes and increase in mitochondrion number, respiration, and basal energy expenditure in adipose tissue relative to control animals. Moreover, the selective SYVN1 inhibitor LS-102 abolished the negative regulation of PGC-1ß by SYVN1 and prevented weight gain in mice. Thus, SYVN1 is a novel post-translational regulator of PGC-1ß and a potential therapeutic target in obesity treatment.

Cell adhesion and glistening formation in hybrid copolymer intraocular lenses.

The purpose of this study is to investigate the cell adhesion and glistening formation properties of various foldable intraocular lenses (IOLs) in vitro. Three conventional hydrophobic methacrylate acrylic (MA) IOLs, a hydrophilic hydroxyethyl methacrylate (HEMA) IOL and a hybrid MA/HEMA copolymer IOL were investigated for immunologically activated cell adhesion and for formation of glistenings resulting from cavitation, by analysis of digital images using NIH Image J PC software. The MA IOLs exhibited a low level of adhering cells but a high level of glistening formation, the HEMA IOL exhibited the reverse tendency, and the MA/HEMA IOL exhibited a low level of both, thus indicating that hybrid MA/HEMA IOLs are less susceptible than HEMA IOLs to cell adhesion and less susceptible than MA IOLs to glistening formation.

Presumed Vogt-Koyanagi-Harada disease with unilateral ocular involvement: report of three cases.

AIM: To report three cases of presumed Vogt-Koyanagi-Harada (VKH) disease with unilateral ocular manifestations. METHODS: This retrospective study reviewed the long-term follow-up observations of three patients who attended the uveitis clinic at Tokyo Medical University Hospital. The patients were followed for 5-16 years with systemic clinical, ophthalmologic and laboratory examinations. Ophthalmoscopic findings, extraocular manifestations, visual acuity, and response to corticosteroid administration were evaluated. RESULTS: Three patients had characteristic clinical features of VKH involving only one eye, including diffuse choroiditis, serous retinal detachment, focal areas of delayed choroidal perfusion, multifocal areas of pinpoint leakage, macular oedema, and optic nerve staining. All patients received systemic corticosteroid therapy during the acute phase of the disease. During the follow-up period (5-16 years), all three patients developed sunset-glow fundus and nummular chorioretinal depigmented scars in the affected eye only, as well as systemic complications of deafness, vitiligo, and poliosis. CONCLUSION: The clinical and laboratory features of all three patients were typical of VKH disease except for the unilateral involvement. It is important for ophthalmologists to recognize unilateral VKH disease, even though it is a rare clinical variant of the disease.

[Terminal care for one-person household].

In addition to visitor medical-examination service, Mutsumi-cho clinic (Minami-ku, Yokohama-city) started a home care service in October 2008. The number of one-person elderly house-hold is 8,483 (about 10%) out of 84,399 house-hold in Minami-ku where 200,000 people reside. We report here our experience in the terminal care of one-person elderly person who lives at home. The regional cooperation is indispensable for seeing-off of the end stage elderly patient at home. We (the physician, visiting nurse, care manager and helper were united and performed) spent a total of 10 days for a terminal care of the end stage elder patient.

[STD in the eye].

In this paper, we review sexually transmitted diseases (STD) involving the eye. Recently conjunctivitis due to Chlamydia trachomatis in children and adults is increasing, and that of Neisseria gonorrhoeae resistant to multiple antibiotics has attracted special attention in our country. Syphilis has many ocular manifestations such as keratitis, iridocyclitis, retinochorioiditis, and neuritis, etc. Ocular complications related to HIV infection, including HIV retinopathy, cytomegalovirus retinitis, zoster ophthalmics, and Kaposi s sarcoma in conjunctiva are increasing in Japan. Phthirus pubis infection of the eye lid, and human T-cell lymphotropic virus type 1 (HTLV-1)-associated uveitis are occasionally reported. Furthermore conjunctival tumor associated with human papilloma virus (HPV) infection, acute retinal necrosis(ARN) due to herpes simplex virus type 2 (HSV-2), as well as hepatitis B virus (HVB) and hepatitis C virus (HVC) retinopathy are also mentioned in this review.

Proteomic surveillance of retinal autoantigens in endogenous uveitis: implication of esterase D and brain-type creatine kinase as novel autoantigens.

PURPOSE: Various retinal proteins are newly exposed to immune system in a process of tissue destructive endogenous uveitis. Some of such proteins could be autoantigens that extend the ocular inflammation in human endogenous uveitis. In this study, we aimed to investigate the possibility of such spreading of autoantigens in endogenous uveoretinitis using a proteomic approach. METHODS: Experimental autoimmune uveoretinitis (EAU) was induced in mice by inoculation with a peptide consisting of amino acids 1-20 (GPTHLFQPSLVLDMAKVLLP) of interphotoreceptor retinoid binding protein (IRBP). Six weeks after immunization, the presence of autoantibodies against the retinal proteins in mice with EAU were examined by two-dimensional electrophoresis followed by western blotting (2D-WB). Retinal proteins targeted by the autoantibodies were identified by mass spectrometry (MS) and their autoantigenicity in patients with endogenous uveitis, such as Behcet's disease (BD, n=36), Vogt-Koyanagi-Harada disease (VKH, n=16), and sarcoidosis (n=17) were examined by enzyme-linked immunosorbent assay. RESULTS: Six new candidate autoantigens, which were detected in mice with EAU using 2D-WD were identified by MS as beta-actin, esterase D (EsteD), tubulin beta-2, brain-type creatine kinase (BB-CK), voltage-dependent anion-selective channel protein, and aspartate aminotransferase. Among the patients with endogenous uveitis, 25% of BD and 25% of VKH patients were positive for anti-EsteD antibody, and 25% of VKH and 38.4% of sarcoidosis patients were positive for anti-BB-CK antibody. CONCLUSIONS: Autoantibodies to EsteD and BB-CK produced in EAU-induced mice were also detected in some endogenous uveitis patients, suggesting that these proteins might be autoantigens spreading in a process of endogenous uveoretinitis.

Phenoxazine derivative, 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one suppresses growth of human retinoblastoma cell line Y79 in vitro and in vivo.

The aim of the study was to evaluate the in vitro and in vivo antitumor effects of the 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one (Phx-1) on the human retinoblastoma cell line Y79. The in vitro effects of Phx-1 on cell viability and apoptosis of the human retinoblastoma Y79 cells, were studied by using colorimetric and flow-cytometric methods. The in vivo antitumor effects of Phx-1 on the human retinoblastoma Y79 cells subcutaneously transplanted in BALB/c nude mice were studied, examining the tumor size, the adverse effects on the mice and the histopathological evaluations including hematoxylin and eosin and immunohistochemical staining in the mass of tumors of human retinoblastoma Y79 cells isolated from the mice. Phx-1 suppressed the viability of Y79 cells dose- and time-dependently and induced apoptosis in Y79 cells in vitro. Phx-1 markedly reduced the growth of Y79 cells transplanted into the mice without causing bodyweight loss. Pathological findings of the tumor mass isolated from mice revealed that the tumor of Y79 cells treated with Phx-1 had a decreased mitotic index, decreased expression of Ki67 and p53, no alteration of bcl-2 level and increased caspase-3 activity compared with the the control. Present results suggested that Phx-1 demonstrated antitumor activity against the human retinoblastoma Y79 cells in vitro and in vivo, by inhibiting cell growth and inducing apoptosis. In addition, Phx-1 exerted few adverse side effects on the mice. Phx-1 may be a useful antitumor drug in the treatment of retinoblastoma, which is the most common and serious intraocular malignant tumor.

Effect of gauge thickness on wound-width measurements in microincision cataract surgery.

PURPOSE: To evaluate the effect of gauge thickness on wound-width measurement values in microincision cataract surgery (MICS). SETTING: Kosei Chuo Hospital, Tokyo, Japan. METHODS: For intraocular lens (IOL) implantation, the incision was enlarged with 1 of 2 knives of different widths. Before and after IOL insertion, the wound width was measured with a 0.15 mm thick gauge (F-gauge), which was the same thickness as both types of knife, and a commercially available 0.35 mm thick gauge (A-gauge). RESULTS: In the 2.2 mm incision group, the mean wound width before IOL insertion was 2.20 mm+/-0.03 (SD) measured with the F-gauge and 2.16+/-0.05 mm measured with the A-gauge; the difference was statistically significant (P=.002). The mean wound width after IOL insertion was 2.41+/-0.08 mm using the F-gauge and 2.35+/-0.09 mm using the A-gauge; the difference was statistically significant (P<.0001). In the 2.4 mm incision group, the mean wound width before IOL insertion was 2.39+/-0.04 mm using the F-gauge and 2.31+/-0.06 mm using the A-gauge (P<.0001); the mean wound width after IOL insertion was greater than 2.5 mm in both groups. CONCLUSIONS: In MICS, when the wound-width gauge thickness exceeded the knife thickness, the potential for measurement errors increased. Thus, a similar thickness between the 2 instruments may be preferable.

The additive effects on intraocular pressure of combining nipradilol 0.25% and latanoprost 0.005% ophthalmic solutions: a prospective, randomized, multicenter study.

PURPOSE: To investigate the effectiveness of combining nipradilol 0.25% and latanoprost 0.005% ophthalmic solutions in improving the intraocular pressures (IOPs) in glaucoma patients. METHODS: We divided the 53 patients into two groups, those who had been treated with latanoprost and those who had been treated with nipradilol. We administered to the first group one dose of latanoprost daily for 12 weeks and to the second group one dose of nipradilol daily for 12 weeks. Each group then received both solutions for another 12 weeks; the latanoprost group received nipradilol and the nipradilol group received latanoprost. IOPs were measured at each 4-week visit. RESULTS: In the patients previously treated with latanoprost, the mean IOP was 19.6+/-2.5 mmHg at baseline, and 14.9+/-2.4 mmHg (23.7% reduction) after 12 weeks of latanoprost monotherapy. The addition of nipradilol decreased the IOP to 13.8+/-1.9 mmHg (29.0% reduction). In the group previously treated with nipradilol, the mean IOP was 20.2+/-3.1 mmHg at baseline, and 16.7+/-3.5 mmHg (17.1% reduction) after 12 weeks of nipradilol monotherapy. Addition of latanoprost decreased the IOP to 14.2+/-3.2 mmHg (29.5% reduction). CONCLUSION: Latanoprost and nipradilol are more effective as a combination therapy than each one by itself.

Preoperative disinfection of the conjunctival sac with antibiotics and iodine compounds: a prospective randomized multicenter study.

PURPOSE: To compare disinfection by povidone-iodine (PI) solution with that by polyvinyl alcohol-iodine (PAI) solution in preoperative eyewash and to assess the optimal duration of the preoperative application of 0.5% levofloxacin (LVFX) ophthalmic solution. METHODS: Two-hundred-seventy-two elderly cataract patients who had undergone phacoemulsification aspiration with intraocular lens insertion at 12 clinical facilities in Japan were randomized into three groups: 3-day, 1-day, and 1-h preoperative application of 0.5% LVFX ophthalmic solution. Each LVFX group was further divided into two subgroups; the patients in one subgroup were treated with diluted PI eyewash, and those in the second subgroup with diluted PAI eyewash, immediately before the surgery. Thus, a total of six groups were studied. Conjunctival scrapings were collected four times: before LVFX application, after LVFX application, after the eyewash with iodine compounds, and after the surgery. Samples of the aqueous humor were collected immediately after the surgery. All samples were cultured aerobically and anaerobically. RESULTS: The 3-day application of LVFX had a significantly higher disinfection rate than the 1-day or 1-h applications (P<0.05). The disinfection rate of PI was 78.0%, and that of PAI was 79.4%; PAI was not inferior to PI as a preoperative disinfectant. However, new isolates, mainly Propionibacterium acnes and Staphylococcus epidermidis, were detected after LVFX application and even after use of the eyewash with iodine compounds. CONCLUSION: Three-day administration of LVFX ophthalmic solution and use of an eyewash with diluted iodine compounds is effective for preoperative disinfection. However, complete elimination of bacteria from the conjunctival sac is difficult.

Upregulation of neurotrophic factor-related gene expression in retina with experimental autoimmune uveoretinitis by intravitreal injection of tacrolimus (FK506).

AIM: The current study was designed to determine whether intravitreal injection of tacrolimus (FK506) modulates the gene expression of neurotrophic factor-related molecules in the retina from eyes with induced experimental autoimmune uveoretinitis (EAU) in rats. METHODS: Rats were immunised with interphotoreceptor retinoid binding protein peptide (R14) and given intravitreal injection of tacrolimus on day 12 after immunisation. As control, immunised rats received intravitreal injection of vehicle. On day 15 after immunisation, changes in the genetic programme associated with neuroprotection and inflammatory responses in the retinas from both groups were determined by DNA microarray analyses and confirmed by real-time PCR analyses. RESULTS: The gene expression of inflammatory responses was markedly reduced in tacrolimus-treated eyes. Genes for molecules associated with neuroprotection (oestrogen receptor, erythropoietin receptor, gamma-aminobutyric acid receptor, protein kinase C, glial cell line-derived neurotrophic factor receptor, fibroblast growth factor and neuropeptide Y receptor) were upregulated in the retinas from tacrolimus-treated eyes. CONCLUSIONS: Intravitreal injection of tacrolimus modulated the genes related to neuroprotection in the retina during the ongoing process of EAU. This treatment may be useful for the neuroprotection of retina with severe uveitis as well as for immunosuppression in the uveitic eyes.

Intravitreal injection of Tacrolimus (FK506) suppresses ongoing experimental autoimmune uveoretinitis in Rats.

AIM: To determine whether intravitreal injection of tacrolimus suppresses ongoing experimental autoimmune uveoretinitis (EAU) in rats. METHODS: Rats were immunised with interphotoreceptor retinoid-binding protein peptide (R14) and given an intravitreal injection of tacrolimus on day 12 after immunisation. Intraocular inflammation was assessed by slit-lamp biomicroscopy and histopathological examination. Interferon gamma and tumour necrosis factor alpha protein levels in the ocular tissues were measured. Gene expression of chemokines was determined in ocular tissues by reverse transcription-polymerase chain reaction. To evaluate the systemic effect of intravitreal injection of tacrolimus, delayed-type hypersensitivity was measured by ear swelling. RESULTS: Clinical and pathological scores showed that ocular inflammation of tacrolimus-treated eyes was markedly less than that of vehicle-treated eyes. The amount of interferon gamma and tumour necrosis factor alpha was considerably inhibited in tacrolimus-treated eyes. The gene expression of monocyte chemattractant protein-1 (MCP-1) and regulated upon activation, normal T cell expressed and secreted (RANTES) was markedly reduced in tacrolimus-treated eyes. Delayed-type hypersensitivity responses were not impaired in tacrolimus-treated rats. CONCLUSIONS: Intravitreal injection of tacrolimus was highly effective in suppressing the ongoing process of EAU without any side effects on systemic cellular immunity. This treatment may be useful in the management of patients with severe uveitis.

Cataract surgery using the bimanual phacoemulsification technique with an Accurus system and Mackool microphaco tip.

PURPOSE: To investigate the safety and effectiveness of the Accurus vitreoretinal and phacoemulsification system (Alcon, Inc.) for bimanual phacoemulsification surgery. SETTING: Kosei Chuo Hospital, Tokyo, Japan. METHODS: Phacoemulsification for age-related cataract was performed using a bimanual technique in 55 eyes and a coaxial technique in 31 eyes. The Accurus system with a venturi pump and a sleeveless ultrasonic tip or a Mackool microphaco tip was used in all cases. The 2 techniques were compared. In bimanual phacoemulsification, the 2 tips were evaluated to determine whether either reduced ocular tissue impairment. RESULTS: The mean best corrected visual acuity 1 day after the surgery was 20/20 in both groups. There was no significant difference between the bimanual group and coaxial group except in ultrasound output (bimanual 22%, coaxial 28%; P = .01). Iris impairment was observed only in the bimanual group but was reduced by the use of the Mackool sleeved microphaco tip. CONCLUSION: Bimanual phacoemulsification for cataract surgery using the Accurus system with a venturi pump and a Mackool microphaco tip was safe and effective and may provide a means of performing combined or successive cataract and vitreoretinal surgery.

Three-dimensional optical coherence tomography of granular corneal dystrophy.

PURPOSE: To describe 3-dimensional optical coherence tomography (OCT) findings in a case of granular corneal dystrophy. METHODS: A 1310-nm high-speed swept-source OCT prototype was used to image the 3-dimensional structures of the cornea of a patient with granular corneal dystrophy. RESULTS: Three-dimensional OCT clearly showed multiple highly reflective lesions, corresponding to the deposits in the cornea. Volume of the deposits within the central 1.5 mm of the cornea was 0.11 mm and occupied 2.98% of the corneal volume. CONCLUSIONS: Three-dimensional OCT allows objective and noninvasive assessment of granular corneal dystrophy.

A case of hypertensive keratouveitis with endotheliitis associated with cytomegalovirus.

AIM: While cytomegalovirus is well known as a pathogenic organism of retinitis, especially associated with human immunodeficiency virus infection, there are few reports of anterior uveitis associated with cytomegalovirus. METHODS: The authors present a case of keratouveitis associated with cytomegalovirus. RESULTS: A 70-year-old Japanese man was referred to the authors because of poorly controlled hypertensive keratouveitis in the left eye. The patient had a history of recurrent hypertensive anterior uveitis. At presentation, the corneal stroma was edematous, with Descemet's folds and pigmented keratic precipitates. The anterior chamber angle was depigmented compared to the fellow eye. Even though pupil dilation and posterior synechiae were absent, iris atrophy was not evident. His right eye appeared normal except for moderate cataract. Funduscopy of the left eye was hazy, with the optic disc showing a normal color but poorly defined details, and no apparent exdative retinitis. The best-corrected decimal visual acuity of the right and left eyes was 0.4 and 0.02, respectively. Intraocular pressure was 11 mmHg in the right eye and 35 mmHg in the left, despite maximum medical therapy. Systemic acyclovir and prednisolone for a month did not improve the hypertensive keratouveitis. The aqueous humor was investigated for herpes simplex virus, varicella-zoster virus and cytomegalovirus. Cytomegalovius genome was detected by polymerase chain reaction analysis. Oral valganciclovir rapidly reduced ocular hypertension within a week. CMV DNA disappeared 3 months after the initiation of valganciclovir. CONCLUSION: The authors reported a case of hypertensive keratouveitis with endotheliitis associated with cytomegalovirus.

Epidemiological survey of intraocular inflammation in Japan.

PURPOSE: To report the frequency and trend of intraocular inflammation based on a survey of new ophthalmology patient visits to university hospitals throughout Japan during 2002. METHODS: A questionnaire was sent to the departments of ophthalmology in 110 university hospitals nationwide to survey the total number of new patients who visited the outpatient clinics for the first time between 1 January and 31 December 2002, and also the number of patients diagnosed with intraocular inflammation during this period. RESULTS: The surveys completed by 41 university hospitals were analyzed in this study. During 2002, a total of 151 299 new ophthalmological patients presented at the 41 institutions, and 3060 (2.2%) of the new patients were diagnosed as having intraocular inflammation. The most frequent intraocular inflammatory disease identified was sarcoidosis (13.3%), followed by Vogt-Koyanagi-Harada (VKH) disease (6.7%), Behçet disease (6.2%), bacterial endophthalmitis (3.8%), herpetic iridocyclitis (3.6%), diabetic iritis (1.6%), human leukocyte antigen-B27-associated uveitis (1.5%), acute retinal necrosis (1.3%), ocular toxoplasmosis (1.1%), ocular toxocariasis (1.1%), uveitis associated with human T lymphotropic virus-1 (also known as HAU) (1.1%), and others. Infectious intraocular inflammation accounted for 16% of all uveitis cases. CONCLUSIONS: Through the collaboration of a large number of institutions, some aspects of the epidemiology of intraocular inflammation in Japan were elucidated. However, the disease concept and diagnostic criteria remain ambiguous for a considerable number of diseases within the spectrum of intraocular inflammation, and the possibility that such factors may bias the present findings cannot be denied. In the future, a prospective survey based on well-defined, common diagnostic criteria is required to obtain more precise epidemiological data.

[Translational research with experimental autoimmune uveoretinitis (EAU)].

Experimental autoimmune uveoretinitis (EAU) induced by immunization with retinal antigen (Santigen or interphotoreceptor retinoid-binding protein; IRBP) serves as an animal model of human uveoretinitis. As the first stage, we demonstrated the similarities between EAU and ocular inflammation in Behçet's disease by investigating anti-retinal antibodies, leukocyte migration inhibition by retinal antigen, immunogenic antigens, aberrant functions of neutrophils, and dominant Th1 lymphocyte reaction. From these findings, we verified that EAU, which is not associated with the systemic disorders observed in Behçet's disease, is an appropriate model for translational research targeting ocular inflammation. In the second stage, we set 3 therapeutic strategies for uveitis in Behçet's disease to be conducted in the translational research: (1) intraocular administration of an immunosuppressive drug; (2) inhibition of Th1 lymphocytes; and (3) activation of immunoregulatory cells. In strategy 1, our studies indicated that intravitreal injection of 10 microg of tacrolimus (FK 506) was not harmful to the retina and was predominantly effective in suppressing ongoing EAU in rats. In strategy 2, two approaches were adopted to prevent differentiation of Thl cells. One is anti-cytokine antibody therapy using anti-IL-12 monoclonal antibodies(mAb). The other is blockade of co-stimulatory signals, especially the ICOS-B7RP-1 pathway. Administration of anti-IL-12 mAb at the time of IRBP immunization completely inhibited development of EAU, and antagonistic anti B7RP-1 mAb suppressed the severity of EAU even when administered after development of EAU. In strategy 3, adoptive transfer of antigen presenting cells treated with a neuropeptide (vasoactive intestinal peptide or calcitonin gene-related peptide) or CD 4+ CD 25+ regulatory T cells suppressed EAU. We look forward to the day when therapies that are being developed in our translational research using EAU will become available for treating intraocular inflammation in Behçet's disease.

Expression of costimulatory molecules on human retinoblastoma cells Y-79: functional expression of CD40 and B7H1.

PURPOSE: To examine the expression of various costimulatory molecules on the human retinoblastoma cell line Y-79 and assess the functional roles of selected costimulatory molecules. METHODS: Y-79 cells were incubated in the presence or absence of IFN-gamma, with or without irradiation (100 Gy). Expression of major histocompatibility complex (MHC) class I molecules, MHC class II, CD80, CD86, CD40, CD70, B7H1, B7DC, B7H2, OX40L, and 4-1BBL on Y-79 cells was measured by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometric analysis. The functional role of CD40-mediated interactions in modifying immune responses to Y-79 was assessed in vitro by using recombinant human CD40 ligand (rhCD40L). The costimulatory effect of B7H1-expressing IFN-gamma-treated Y-79 cells on proliferation of purified T cells was studied in Y-79/T-cell coculture experiments with a blocking anti-B7H1 monoclonal antibody (mAb). RESULTS: CD40 and B7H2 were consistently detected on Y-79 cells by RT-PCR and flow cytometry. Cell surface expression of CD40 was upregulated on stimulation by IFN-gamma alone, radiation alone, and IFN-gamma combined with radiation. B7H1 expression was induced by IFN-gamma stimulation and increased further when irradiated Y-79 cells were stimulated by IFN-gamma. Treatment of Y-79 cells with rhCD40L enhanced cell surface expression of MHC class I and intercellular adhesion molecule (ICAM)-1 and also stimulated monocyte chemotactic protein (MCP)-1 production. Proliferative response of purified CD3+ T cells costimulated with IFN-gamma-stimulated Y-79 was significantly enhanced by the addition of anti-B7H1 mAb. CONCLUSIONS: These results suggest that CD40 expressed on Y-79 plays an important role in augmenting antitumor immunity. In contrast, the expression of B7H1 on IFN-gamma-treated Y-79 cells contributes to the suppression of T cells. The dual effects of CD40 and B7H1 on Y-79 cells may contribute to positive or negative regulation of antitumor immune responses in human retinoblastoma.

Induction of eye-derived tolerance does not depend on naturally occurring CD4+CD25+ T regulatory cells.

PURPOSE: Regulatory CD4+ T cells (T regs) arise in the spleens of mice with anterior chamber-associated immune deviation (ACAID), an eye-derived tolerance evoked by injection of antigen into the ocular anterior chamber (AC). The current study was conducted to investigate the possibility that these T regs express CD25 and are derived from natural CD4+CD25+ T cells. METHODS: Naïve T cells from DO11.10 mice were activated in vitro by ovalbumin (OVA)-pulsed, TGFbeta-treated antigen-presenting cells (APCs), and the expression of CD25 assayed by flow cytometry. OVA-specific ACAID T regs were obtained from the spleens of DO11.10 mice with ACAID to OVA. Immunomagnetic enrichment was used to sort out CD4+CD25+, and CD4+CD25- ACAID T cells before they were injected into OVA-immunized mice or examined for mRNA expression of the regulatory T-cell transcription factor Foxp3. In addition, before AC injection of OVA, systemic depletion of CD25+ T cells was performed with injections of anti-IL-2 receptor antibody into the mice. RESULTS: OVA-specific T cells from DO11.10 mice expressed CD25 when exposed to OVA-pulsed, TGFbeta-treated APCs, even when the DO11.10 T cells were depleted of CD25+ cells before their in vitro stimulation. In addition, DH was suppressed in naïve mice that were injected with CD4+CD25+ or CD4+CD25- ACAID T cells. The CD4+CD25+, but not the CD4+CD25-, ACAID T regs expressed Foxp3. Finally, OVA induced ACAID in mice depleted of CD25+ cells. CONCLUSIONS: Some of the CD4+ T regs of ACAID arise from CD25- precursors, and the induction of ACAID is not dependent on the presence of natural CD4+CD25+ T regs.

Elevated serum levels of CXCL9/monokine induced by interferon-gamma and CXCL10/interferon-gamma-inducible protein-10 in ocular sarcoidosis.

PURPOSE: Sarcoidosis is a chronic multisystem granulomatous disorder characterized by an accumulation of activated CD4+ T cells and monocytes/macrophages in involved organs. Chemokines are required for the extravasation of leukocytes to the inflammation site. This study was undertaken to determine which chemokines are augmented in the serum of patients with active ocular sarcoidosis. METHODS: Seventeen patients with diagnosed ocular sarcoidosis, 28 with suspected ocular sarcoidosis, 16 with Behçet's disease, 17 with Vogt-Koyanagi-Harada disease, and 18 healthy subjects were studied. Serum levels of CCL2, CCL5, CXCL8, CXCL9, and CXCL10 were simultaneously measured by cytometric bead array using flow cytometer. In addition, serum CXCL9 and CXCL10 levels in the patients with diagnosed or suspected ocular sarcoidosis were compared with respect to ocular disease activity, the presence of bilateral hilar lymphadenopathy (BHL), and laboratory data. RESULTS: Serum levels of both CXCL9 and CXCL10 were markedly elevated in the patients with diagnosed or suspected ocular sarcoidosis compared with patients with other types of uveitis and healthy subjects. Although CCL2 and CXCL8 were detected in the serum of all subjects, the levels were extremely low with no significant differences between groups. Elevation of serum CXCL9 and CXCL10 in ocular sarcoidosis correlated significantly with ocular disease activity and ACE (angiotensin converting enzyme) levels and was unrelated to the presence of BHL, erythrocyte sedimentation rate, white blood cell count, serum IgG, or serum lysozyme. CONCLUSIONS: The results demonstrated that serum levels of CXCL9 and CXCL10 were elevated markedly in the patients with ocular sarcoidosis and correlated with ocular disease activity and ACE level.