Molecular Signatures of Natural Killer Cells in CMV-Associated Anterior Uveitis, A New Type of CMV-Induced Disease in Immunocompetent Individuals.

Cytomegalovirus (CMV) causes clinical issues primarily in immune-suppressed conditions. CMV-associated anterior uveitis (CMV-AU) is a notable new disease entity manifesting recurrent ocular inflammation in immunocompetent individuals. As patient demographics indicated contributions from genetic background and immunosenescence as possible underlying pathological mechanisms, we analyzed the immunogenetics of the cohort in conjunction with cell phenotypes to identify molecular signatures of CMV-AU. Among the immune cell types, natural killer (NK) cells are main responders against CMV. Therefore, we first characterized variants of polymorphic genes that encode differences in CMV-related human NK cell responses (Killer cell Immunoglobulin-like Receptors (KIR) and HLA class I) in 122 CMV-AU patients. The cases were then stratified according to their genetic features and NK cells were analyzed for human CMV-related markers (CD57, KLRG1, NKG2C) by flow cytometry. KIR3DL1 and HLA class I combinations encoding strong receptor-ligand interactions were present at substantially higher frequencies in CMV-AU. In these cases, NK cell profiling revealed expansion of the subset co-expressing CD57 and KLRG1, and together with KIR3DL1 and the CMV-recognizing NKG2C receptor. The findings imply that a mechanism of CMV-AU pathogenesis likely involves CMV-responding NK cells co-expressing CD57/KLRG1/NKG2C that develop on a genetic background of KIR3DL1/HLA-B allotypes encoding strong receptor-ligand interactions.

Guidance on Noncorticosteroid Systemic Immunomodulatory Therapy in Noninfectious Uveitis: Fundamentals Of Care for UveitiS (FOCUS) Initiative.

TOPIC: An international, expert-led consensus initiative to develop systematic, evidence-based recommendations for the treatment of noninfectious uveitis in the era of biologics. CLINICAL RELEVANCE: The availability of biologic agents for the treatment of human eye disease has altered practice patterns for the management of noninfectious uveitis. Current guidelines are insufficient to assure optimal use of noncorticosteroid systemic immunomodulatory agents. METHODS: An international expert steering committee comprising 9 uveitis specialists (including both ophthalmologists and rheumatologists) identified clinical questions and, together with 6 bibliographic fellows trained in uveitis, conducted a Preferred Reporting Items for Systematic Reviews and Meta-Analyses protocol systematic review of the literature (English language studies from January 1996 through June 2016; Medline [OVID], the Central Cochrane library, EMBASE, CINAHL, SCOPUS, BIOSIS, and Web of Science). Publications included randomized controlled trials, prospective and retrospective studies with sufficient follow-up, case series with 15 cases or more, peer-reviewed articles, and hand-searched conference abstracts from key conferences. The proposed statements were circulated among 130 international uveitis experts for review. A total of 44 globally representative group members met in late 2016 to refine these guidelines using a modified Delphi technique and assigned Oxford levels of evidence. RESULTS: In total, 10 questions were addressed resulting in 21 evidence-based guidance statements covering the following topics: when to start noncorticosteroid immunomodulatory therapy, including both biologic and nonbiologic agents; what data to collect before treatment; when to modify or withdraw treatment; how to select agents based on individual efficacy and safety profiles; and evidence in specific uveitic conditions. Shared decision-making, communication among providers and safety monitoring also were addressed as part of the recommendations. Pharmacoeconomic considerations were not addressed. CONCLUSIONS: Consensus guidelines were developed based on published literature, expert opinion, and practical experience to bridge the gap between clinical needs and medical evidence to support the treatment of patients with noninfectious uveitis with noncorticosteroid immunomodulatory agents.

Pan-viral Antibody Repertoire of Aqueous Humor in Cytomegalovirus Uveitis.

PURPOSE: The identification of infectious etiologies is important in the management of uveitis. Ocular fluid testing is required, but multiplex testing faces challenges due to the limited volume sampled. The determination of antibody repertoire of aqueous humor (AH) is not possible with conventional assays. We investigated the use of a highly multiplexable serological assay VirScan, a Phage ImmunoPrecipitation Sequencing (PhIP-Seq) library derived from the sequences of more than 200 viruses to determine the antibody composition of AH in patients with uveitis. DESIGN: Prospective, case control study. METHODS: We analyzed the paired AH and plasma samples of 11 immunocompetent patients with active polymerase chain reaction-positive cytomegalovirus (CMV) anterior uveitis and the AH of 34 control patients undergoing cataract surgery with no known uveitis in an institutional practice. The samples were tested using VirScan PhIP-Seq, and the entire pan-viral antibody repertoire was determined using peptide tile ranking by normalized counts to identify significant antibodies enrichment against all viruses with human tropism. RESULTS: Significant enrichment of antibodies to Herpesviridae, Picornavirdae, and Paramyxoviridae was detectable in 20 µL of AH samples from patients with CMV uveitis and controls. Patients with CMV uveitis had relative enrichment of anti-CMV antibodies in AH compared with their plasma. Epitope-level mapping identified significant enrichment of antibodies against CMV tegument protein pp150 (P = 1.5e-06) and envelope glycoprotein B (P = .0045) in the AH compared with controls. CONCLUSIONS: Our proof-of-concept study not only sheds light on the antibody repertoire of AH but also expands the utility of PhIP-Seq to future studies to detect antibodies in AH in the study of inflammatory eye diseases.

Role of Optical Coherence Tomography in the Prognosis of Vogt-Koyanagi-Harada Disease.

Purpose: To evaluate the prognostic value of OCT in acute VKH diseaseMethods: Retrospective series of consecutive VKH patients seen with the first presentation, a minimum follow-up of 1 year and serial OCT imaging. The primary outcome was the clinical activity phase at 1, 2 and 3 years.Results: Among 29 subjects, 7 resolved, 17 became chronic, and 5 developed chronic recurrent VKH. Median RNFL measurement obtained 2 months post-treatment was higher among subjects who developed chronic VKH (p-value = 0.03) and chronic recurrent disease (p-value = 0.04). Median CRT 1-week post-treatment was lower among subjects who developed PPA (p-value = 0.03). CRT 2 months post-treatment was higher among subjects who developed vitiligo (p-value = 0.02).Conclusion: OCT is helpful in early recognition of the predominantly optic disc swelling variant which carries a poorer prognosis. Lower CRT reading at 1 week and persistently raised CRT at 2 months post-treatment correlated with chronic recurrent disease.

Singapore Ocular Tuberculosis Immunity Study (SPOTIS): Role of T-lymphocyte Profiling in Patients with Presumed Ocular Tuberculosis.

Objective: A prospective clinical study to assess the utility of CD4 + T cell lymphocyte profiling from peripheral blood in patients with ocular tuberculosis (TB).Methods: Thirty-six Asian patients with presumed diagnosis of ocular TB were recruited for T-lymphocyte profiling. MTB antigen specific CD4 assay was set up, and flow cytometric data were analyzed using FlowJo software.Results: There was no significant difference between treatment responders and non-responders for the proportion of CD4 + T cells specific for PPD or ESAT-6+ CFP-10, but treatment responders did have significantly higher frequency of CD38+ (p = .0357) and CD38+ HLA-DR+ (p = .0357) on the PPD-specific CD4 + T cells.Conclusion: This study is one of the first of its kind to look into MTB specific T cell activation marker profiling of peripheral blood in patients with ocular TB. Further studies need to be undertaken to assess the utility of CD4 + T cell phenotypes as a biomarker for ocular TB.

Kinetics of Tear Fluid Proteins after Endothelial Keratoplasty and Predictive Factors for Recovery from Corneal Haze.

Endothelial keratoplasty (EK) is less invasive with faster recovery as compared to conventional penetrating keratoplasty, however, it relies on the clarity of the host corneal stroma. Corneal transplantation involves the induction of immune tolerance for allogeneic tissues as well as the corneal wound healing process, in which coordinated interactions between cytokines and growth factors are critical. In this study, we profiled the expression of 51 soluble factors in the tear fluid over the course of EK and have provided evidence of dynamic changes in cytokine expression in the ipsilateral and contralateral eyes. Cluster analyses classified the cytokine expression kinetics into five groups. Group 1 proteins included TGF-b1, IL-1b, and innate proinflammatory cytokines, which bilaterally increased after surgery, despite the use of topical corticosteroid in the transplanted eyes. Local corticosteroids suppressed cytokines involved in adaptive immunity in the transplanted eyes but not in the contralateral eyes. We found tear protein expression at baseline and one week post-surgery to be a potential predictive biomarker of delayed recovery after EK in terms of the corneal haze and visual acuity. Furthermore, Group 1 tear proteins were most associated with persistent corneal haze pre-surgery as well as visual acuity at one month-post transplant.

Cytomegalovirus Anterior Uveitis Following Topical Cyclosporine A.

Cytomegalovirus (CMV) anterior uveitis and endotheliitis occurs among immunocompetent individuals and may manifest as Posner-Schlossman syndrome or Fuchs uveitis syndrome. The condition may first present following ophthalmic surgery, the use of a fluocinolone-sustained steroid drug delivery implant, or the use of topical prostaglandin analogues for the treatment of glaucoma. We report the first case of a non-human immunodeficiency virus-infected individual who presented with CMV anterior uveitis after the use of topical cyclosporine A 0.05% ophthalmic emulsion for the treatment of symptomatic dry eyes.

Multimodal Imaging in Anterior Uveitis.

Anterior uveitis is the most common group of uveitis worldwide, with a diverse spectrum ranging from autoimmune and infectious to masquerade etiologies. Elucidation of the underlying etiology may be challenging, but it remains important, especially for recurrent uveitis. Multimodal imaging has improved our understanding and management of many posterior uveitis and panuveitis. Similarly, a wide variety of anterior segment ophthalmic imaging techniques are available to allow the monitoring of the structural changes that may develop during anterior uveitis. These include anterior segment photography, specular microscopy, in vivo confocal microscopy, anterior segment optical coherence tomography (AS-OCT), ultrasound biomicroscopy (UBM), laser flare photometry, and less commonly used techniques such as anterior segment fluorescein and indocyanine green angiography, iris autofluorescence, and infrared imaging. This review recapitulates the past and recent application of these ophthalmic imaging modalities to the diagnosis and management of anterior uveitis.

The Pattern of Uveitis in Sri Lanka.

PURPOSE: To describe the pattern of uveitis at a tertiary uveitis referral clinic in Sri Lanka. METHODS: Consecutive charts identified from a registry between January 2010 and December 2014 at the Mediclinic Hospital, Colombo were retrospectively reviewed. RESULTS: Among 750 patients, anterior uveitis (AU) was the most common (285, 38%), followed by posterior uveitis (187, 25%), intermediate uveitis (150, 20%), and panuveitis (128, 17%). 485 (65%) were idiopathic. The top identified causes of AU were seronegative-spondyloarthropathy-related-AU (37, 13%), HLA-B27-related-AU without systemic associations (25, 9%), herpetic-AU (18, 6%), and trematode uveitis (8, 3%). The main posterior uveitis were toxoplasmosis (34, 18%), tuberculosis (21, 11%), and sarcoidosis (17, 9%). 78% of intermediate uveitis were idiopathic, with 12 (8%) tuberculosis, and 10 (7%) sarcoidosis. The most common panuveitis were sarcoidosis (18, 14%), tuberculosis (14, 11%), and Vogt-Koyanagi-Harada disease (10, 8%). Among children (≤12 years) accounting for 25 (3%) of cases, the most common presentations were idiopathic intermediate uveitis (7, 28%), idiopathic-AU (4, 16%), toxoplasmosis (4, 16%), trematode uveitis (3, 12%), and toxocariasis (2, 8%). CONCLUSIONS: An infectious etiology must be considered in posterior uveitis and uveitis among children in the Sri Lankan population.

The Spectrum of Uveitis in Southern Vietnam.

PURPOSE: To describe the pattern of uveitis among Vietnamese at two eye hospitals in Southern Vietnam. METHODS: We retrospectively reviewed the charts of 212 consecutive uveitis cases that presented to two eye hospitals in Ho Chi Minh City, Vietnam, from July 2011 to February 2015. The patients were identified from a database maintained by the hospitals. Patients with keratitis, episcleritis, orbital inflammation, post-surgical endophthalmitis, traumatic iritis, and corneal graft rejection were excluded. Data collected included demographic, clinical, and laboratory findings. RESULTS: Uveitis was seen most commonly in the 21-60-year-old age group. Gender distribution was not significantly different among the various age groups. Anterior uveitis was the most common (46%), followed by posterior uveitis (22%), panuveitis (18%), and intermediate uveitis (14%). Infectious etiologies were observed in 27%. Idiopathic uveitis (36%) was the most common non-infectious uveitis, followed by Vogt-Koyanagi-Harada disease (14%) and Behçet disease (7%). Tuberculous uveitis was the most common infectious etiology (9%), followed by toxocariasis (6%) and herpetic uveitis (6%), and cytomegalovirus anterior uveitis (4%). CONCLUSIONS: In southern Vietnam, infectious uveitis, such as tuberculosis, toxocariasis, and herpetic infection, are common but toxoplasmosis is rare. Idiopathic uveitis, Vogt-Koyanagi-Harada disease, and Behçet disease are the most common non-infectious uveitis, with a notable absence of birdshot retinochoroidopathy.

The Pattern of Uveitis among Chinese, Malays, and Indians in Singapore.

PURPOSE: To describe the pattern of uveitis among Chinese, Malays, and Indians at a tertiary referral institution in Singapore. METHODS: Charts between January 1997 and December 2010 were retrospectively reviewed. Charts between January 2014 and December 2014 were prospectively reviewed. RESULTS: A total of 1249 and 148 charts were retrospectively and prospectively reviewed, respectively. The top causes of anterior uveitis (AU) were HLA-B27, idiopathic, and CMV AU. The top known causes of intermediate uveitis were tuberculosis, primary intraocular lymphoma, and sarcoidosis. The top causes of posterior uveitis were CMV retinitis, toxoplasmosis, and dengue maculopathy. The top causes of panuveitis were VKH, idiopathic panuveitis, tuberculosis, and Behçet disease. HLA-B27 and CMV AU were more frequent among Chinese (21% vs 9% (non-Chinese); p<0.001; 10% vs 5% (non-Chinese); p<0.001, respectively). Tuberculous uveitis was more frequent among Malays and Indians (12% (non-Chinese) vs 5% (Chinese), p<0.001). CONCLUSIONS: Different uveitis patterns were encountered among patients of different races.

Anterior Chamber Angle and Intraocular Pressure Changes After Phacoemulsification: A Comparison Between Eyes With Closed-angle and Open-angle Glaucoma.

PURPOSE: To compare the angle and intraocular pressure (IOP) changes after phacoemulsification between eyes with closed-angle or open-angle glaucoma. METHODS: Angle measurements using Visante AS-OCT imaging was performed for a prospective cohort of 24 subjects with closed-angle and 30 subjects with open-angle glaucoma before and 3 months after phacoemulsification. IOP measurement was measured at 6 and 12 months after surgery using Goldmann applanation tonometry as secondary outcome measures. RESULTS: Eyes with closed angles were smaller than those with open angles (mean axial length 22.88 vs. 24.11 mm, P<0.001). Mean anterior chamber depth, area, volume, AOD500, AOD750, ARA, TISA500, and TISA750 increased after phacoemulsification in all eyes regardless of preexisting angle status (all P<0.001). Increase in AOD500, AOD750, TISA500, and TISA750 were greater in eyes with open angles compared with closed angles (P=0.03, 0.04. 0.04, 0.04, respectively). Mean IOP decreased by 1.8 and 2.1 mm Hg at 6 and 12 months, respectively, after phacoemulsification for all eyes (P<0.001 for both timepoints compared with preoperative baseline). However, postoperative reduction in the mean IOP was not significantly different between eyes with closed and open angles (Mann-Whitney test P=0.32 at 6 mo and P=0.75 at 12 mo postsurgery compared with preoperative). CONCLUSIONS: Angle opening postphacoemulsification was considerable in all eyes. A similar IOP reduction after phacoemulsification was observed in all eyes regardless of angle status.

Prevalence and risk factors of meibomian gland dysfunction: the Singapore Malay eye study.

PURPOSE: To describe the prevalence and associations of meibomian gland dysfunction (MGD) in an urban Malay population in Singapore. METHODS: Population-based cross-sectional study of 3280 (78.7% response rate) Malay persons aged 40 to 80 years, living in Singapore. MGD was defined by a slit-lamp clinical examination as either lid margin telangiectasia or meibomian gland orifice plugging in at least one eye. Participants underwent a standardized questionnaire and clinical examination, including laboratory investigations. Data were analyzed for 3271 persons. RESULTS: The age-standardized prevalence of MGD was 56.3% [95% confidence interval (CI), 53.3-59.4]. A higher MGD prevalence was found in male participants across all age groups [odds ratio (OR), 1.30; 95% CI, 1.35-1.79], postmenopausal women (OR, 1.64; 95% CI, 1.19-2.33), and all participants with pinguecula (OR, 2.43; 95% CI, 2.08-2.85), high diastolic blood pressure (OR, 1.32; 95% CI, 1.08-1.62), and use of angiotensin II receptor blockers (OR, 4.02; 95% CI, 1.74-9.27). CONCLUSIONS: MGD was highly prevalent in this Asian population and associated with various systemic and ocular conditions.

The nuclear-factor kappaB pathway is activated in pterygium.

PURPOSE: Pterygium is a prevalent ocular surface disease with unknown pathogenesis. The authors investigated the role of nuclear factor kappa B (NF-κB) transcription factors in pterygium. METHODS: Surgically excised primary pterygia were studied compared with uninvolved conjunctiva tissues. NF-κB activation was evaluated using Western blot analysis, ELISA, and DNA-binding assays. Primary pterygium fibroblasts were treated with TNF-α (20 ng/mL), and NF-κB activation was evaluated using immunocytochemistry, Western blot analysis, phospho-IκBα ELISA, and DNA-binding assays. TNF-α stimulation of NF-κB target genes RelB, NFKB2, RANTES, MCP-1, ENA-78, MMP-1, MMP-2, and MMP-3 in pterygium fibroblasts was compared with that in primary tenon fibroblasts by real-time PCR. RESULTS: Phosphorylation of IκBα (Ser32) was increased in pterygia tissues compared with uninvolved conjunctiva tissues, as determined by Western blot analysis and ELISA. IκBα expression was decreased, whereas nuclear RelA and p50 DNA-binding capacities were increased. Within 30 minutes of treatment with TNF-α, pterygium fibroblasts showed increased IκBα phosphorylation and nuclear translocation of RelA and p50. Treatment with TNF-α beyond 12 hours resulted in increased nuclear expression of RelB, p100, and p52. Furthermore, the upregulation of RANTES, MCP-1, ENA-78, MMP-1, MMP-2, and MMP-3 expression was more pronounced in TNF-α-treated pterygium fibroblasts than in tenon fibroblasts. CONCLUSIONS: The NF-κB pathway is shown for the first time to be activated in pterygia tissues compared with normal conjunctiva tissues. Stimulation by the inflammatory cytokine TNF-α can activate both canonical and noncanonical NF-κB pathways in pterygium fibroblasts with concomitant upregulation of NF-κB target genes.