Acute Anterior Uveitis Patients Research Articles

Comprehensive single-cell profiling of monocytes in HLA-B27-positive ankylosing spondylitis with acute anterior uveitis.

Acute anterior uveitis (AAU) is a common extra-articular manifestation of ankylosing spondylitis (AS), particularly in patients positive for the human leucocyte antigen (HLA)-B27 genetic marker. To explore the underlying mechanisms of HLA-B27+ AS-associated AAU, we employed single-cell RNA sequencing to profile the transcriptomes of peripheral blood mononuclear cells in three HLA-B27+ AS-associated AAU patients and three healthy controls (HCs). We identified 11 distinct immune cell clusters, with a particular focus on monocytes, revealing six subsets, including three previously unidentified subsets, namely, GTPase immune-associated proteins, Th17-related, and lncRNA monocytes, with unique gene expression patterns. Significant differences in monocyte composition, activation states, and gene expression were observed between patients and HCs, particularly within HLA monocyte subpopulations. Notably, enhanced expression of X-inactive specific transcript and myeloid cell nuclear differentiation antigen genes was validated across monocyte subclusters in patients. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis highlighted significant enrichment in antigen processing and presentation pathways, shedding light on the disease's molecular mechanisms. These findings provide novel insights into the molecular mechanisms of HLA-B27+ AS-associated AAU and may contribute to the development of targeted diagnostic and therapeutic strategies. Further clinical validation is essential.

Frequency of HLA-B27 Haplotype Among Patients with Acute Anterior Uveitis and Controls in Ethiopia

ABSTRACT Purpose We compared the distribution of the HLA-B27 allele among Ethiopian patients with acute anterior uveitis (AAU) and controls without that disease. Methods The clinical features of patients were collected from their medical records. HLA-B27 genotyping was performed for 64 patients, with AAU using the real-time polymerase chain reaction (RT-PCR), and the results were compared to those from a panel of 192 healthy, unrelated volunteer control participants (refraction patients and volunteers) free of signs of anterior uveitis. Other outcomes were assessed comparing HLA-B27 positive vs. negative patients. Results The histocompatibility antigen HLA-B27 was identified in 6 out of 64 (9.4%) AAU patients, compared with 1.0% in controls (Odds Ratio = 9.8, 95% CI (1.93, 50.01)). Sub-group analysis of the cases revealed that patients with HLA-B27 positivity had a higher incidence of ocular hypertension (Odds Ratio = 5.93, 95% CI (1.29, 27.2)). Conclusion The antigen HLA-B27 was represented ~10-fold more frequently in the patient group than in the control group. HLA-B27 allele distribution in both cases and controls is lower than in reports from Caucasian and Asian populations, but similar to South Africa.

Differential Composition and Structure of the Microbiota from Active and Inactive Stages of HLA-B27-associated Uveitis by Paired Fecal Metagenomes

ABSTRACT Purpose To compare the diversities and abundances of bacterial taxa in the microbiome of patients with HLA B27-positive acute anterior uveitis (AAU) in the active and inactive phases. Methods An observational descriptive prospective and comparative study was conducted in ten HLA-B27-positive AAU patients (44.6 ± 13.4 years). The microbiome of the stool samples obtained in the active and inactive stages was analyzed by sequencing the V3 region of the 16S rRNA gene. Results The differences in the bacteria profile between active and inactive stages in each individual were confirmed (p < 0.0001). Ten OTUs were found exclusively in the active phase of 90% of the individuals, suggesting a proinflammatory association. Blautia OUT_4 and Faecalibacterium OUT_2 abundances showed a direct relationship between abundance and severity of ocular inflammation. Two OTUs were exclusive of the inactive stage, suggesting an anti-inflammatory role. Conclusion The metagenomic profile of the fecal microbiota differs in the acute phase of the AAU compared to when the inflammation subsides, despite being the same individual and a short time-lapse. AAU is a fertile field for studying the connection between subtle rapid changes in microbiota and their systemic consequences.

Cellular indexing of transcriptomes and epitopes by sequencing (CITE-Seq) of peripheral blood to dissect the effect of different spondyloarthropathies

Abstract Spondyloarthritis is a family of immune mediated inflammatory disorders with overlapping articular and extraarticular clinical manifestations and a strong association to human leukocyte antigen (HLA)-B27. Despite a strong association of HLA-B27 with prototypic spondyloarthropathies such as axial spondyloarthritis (axSpA) and acute anterior uveitis (AAU), the underlying mechanisms are not known. Here, we aim to determine whether HLA-B27 associated axSpA and AAU are associated with common or divergent immunological markers in the blood. Previously, we did single cell RNA-seq analysis on a small number of patients with AAU and axSpA, which revealed increase in T cell, MAIT cells and NK cell clusters in AAU patients, whereas axSpA patients had increase in MAIT cells, gd-T cells, ILCs and Tregs and decrease in naïve CD8 T cell clusters, suggesting that distinct signatures maybe associated with axSpA and AAU. Changes in ILCs or MAIT cells may suggest involvement of the gut mucosal surface in pathogenesis. This is consistent with our previous studies showing gut microbial perturbation in clinical samples and in rat model of spondyloarthritis. Leveraging on our preliminary data, we are performing CITE-seq to capture paired transcriptome and cell surface epitope expression in individual PBMCs from 5 subjects each with axSpA, AAU and axSpA+AAU in comparison with healthy individuals (5 HLA-B27 negative and 5 HLA-B27 positive). Since, HLA-B27 allele is also present in healthy population, these analyses will allow us to dissect the effect of host genetics from the disease. These studies represent potential advances in the understanding of spondyloarthritis and may reveal genetic and disease biomarkers associated with axSpA and AAU. TG (principal investigator) is a recipient of research awards from the Spondylitis Association of America, SPARTAN (Spondyloarthritis Research and Treatment Network), GRAPPA (Group for Research and Assessment of Psoriasis and Psoriatic Arthritis), The Collins Medical Trust, Medical Research Foundation of Oregon and, Innovation Award from OHSU. This work was funded in part by the Grandmaison Fund for Autoimmunity Research. JTR is a recipient of the Innovative Research Award, Rheumatology Research Foundation.

Association of TBX21 gene polymorphisms and acute anterior uveitis risk in a Chinese population: A case-control study

Copy number variations (CNVs) in TBX21 gene have been reported to be significantly and positively correlated with acute anterior uveitis (AAU). Our study was performed to further determine whether single nucleotide polymorphisms (SNPs) in TBX21 gene confer susceptibility to AAU in a Chinese population. In our case-control study, 420 AAU patients and 918 healthy controls were included. SNP genotyping was conducted via the MassARRAY™ iPLEX Gold platform. Association and haplotype analyses were performed via SPSS 23.0 and SHEsis software. No significant association was observed between two candidate SNPs of TBX21 gene (rs4794067, rs11657479) and susceptibility to AAU (Pc > 0.05). In stratification analysis, the result also showed no significant difference between the HLA-B27 positive AAU patients and non-typed healthy controls. Additionally, no association was detected between TBX21 haplotypes and AAU risk. In conclusion, the polymorphisms rs4794067 and rs11657479 in TBX21 gene did not confer disease susceptibility to AAU in a Chinese population.

Keep an Eye on the Back: Spondyloarthritis in Patients With Acute Anterior Uveitis.

This study was undertaken to analyze the prevalence of spondyloarthritis (SpA) in patients with acute anterior uveitis (AAU), to identify parameters associated with the presence of SpA, and to evaluate the performance of referral algorithms for identifying patients with a high probability of having SpA. Prospectively recruited consecutive patients with noninfectious AAU underwent structured rheumatologic assessment including magnetic resonance imaging of the sacroiliac joints, allowing a definitive diagnosis/exclusion of concomitant SpA. Fisher's exact test and Mann-Whitney U test were used to compare AAU patients with SpA and AAU patients without SpA. Furthermore, logistic regression analyses were performed. The predictive performance of SpA referral strategies was analyzed by calculating the sensitivity, specificity, positive predictive value, and positive and negative likelihood ratios. Among the 189 AAU patients evaluated, 106 (56%) were diagnosed as having SpA. The majority of SpA patients (93%) had predominantly axial SpA and 7 patients had peripheral SpA. In 74 patients (70%), the SpA diagnosis was established for the first time. In multivariable logistic regression analysis, psoriasis (odds ratio [OR] 12.5 [95% confidence interval (95% CI) 1.3-120.2]), HLA-B27 positivity (OR 6.3 [95% CI 2.4-16.4]), elevated C-reactive protein level (OR 4.8 [95% CI 1.9-12.4]), and male sex (OR 2.1 [95% CI 1.1-4.2]) were associated with the presence of SpA. None of the ophthalmologic parameters were found to be predictive of SpA. The Dublin Uveitis Evaluation Tool (DUET) showed higher specificity for SpA recognition than the Assessment of SpondyloArthritis international Society (ASAS) tool for the early referral of patients with a suspected diagnosis of axial SpA (specificity for SpA 42% versus 28%), whereas the sensitivity of the ASAS tool was slightly higher than the DUET tool (sensitivity for SpA 80% versus 78%). However, more than 20% of the AAU patients in this study who were diagnosed as having SpA would have been missed by both referral strategies. Our study revealed a high prevalence of SpA in AAU patients overall, as well as a high prevalence of previously undiagnosed SpA in AAU patients. Therefore, we propose rheumatologic evaluation for all AAU patients with musculoskeletal symptoms.

Epidemiology and characteristics of common forms of anterior uveitis at initial presentation in a tertiary facility in Japan.

To elucidate detailed epidemiological profile of common types of anterior uveitis (AU) in real-world clinical setting of a tertiary facility in Japan, and to evaluate the characteristic clinical findings at initial presentation. Retrospective cohort study. Clinical charts of 275 patients (335 eyes) aged 52.5 ± 19.1 years were reviewed retrospectively. Herpetic AU was diagnosed by multiplex polymerase chain reaction tests using aqueous humor. Time of uveitis onset, gender, laterality, disease course since the initial onset of AU, visual acuity (VA) and intraocular pressure (IOP) at first visit, and definitive diagnosis were collected from clinical charts. Acute AU (AAU) was the most common (21.8%) form of AU; followed by herpetic AU (20.7%) comprising Herpes Simplex Virus (HSV) (8.0%), Varicella Zoster Virus (VZV) (9.1%) and cytomegalo virus (CMV) (3.6%); scleritis (13.5%); diabetic iritis (7.6%), and Posner-Schlossman syndrome (5.5%). Unilateral AU constituted 78.2%, and VA less than 20/30 accounted for 31.2%. Of all the eyes, 16.1% had an IOP higher than 20 mmHg, out of which 37.0% had herpetic AU, followed by scleritis in 25.9%, and Posner-Schlossman syndrome (PSS) in 11.1%. AU patients over 60 years of age were 40.4%, in which 34.2% had herpetic AU, followed by scleritis in 14.4% and AAU in 13.5%. Herpetic AU patients were significantly older and had higher IOP compared with AAU patients. The most frequent AU was AAU, followed by herpetic AU. Herpetic AU patients were older and had higher intraocular pressure than AAU patients, although VA was equally impaired in both groups.

Assessment of the Anti-inflammatory Effects of NORFLO® ORO in Acute Relapses of HLA-B27-associated Autoimmune Uveitis: A Multicenter, Randomized, Placebo-controlled, Double-blind Clinical Study

ABSTRACT Background An effective therapy to reduce the number and severity of HLA-B27-related acute anterior uveitis (AAU) recurrences represents a clinical need. Curcumin is a promising therapeutic option in various inflammatory eye diseases. To enhance its absorption and eye tissue selectivity, a phospholipidic-curcumin complex (PHBC) has been formulated (Iphytoone®, Eye Pharma S.p.A.). Aims This study investigates if PHBC is effective and safe to decrease the number and intensity of HLA-B27-related AAU relapses. Methods HLA-B27-related AAU patients were randomly divided to receive PHBC or placebo for 12 months (NCT03584724). Results Compared with the previous year, the number of relapses decreased in both groups. The proportion of responders was significantly higher in the PBHC group. The severity of attacks was comparable. The study drug was well tolerated. Conclusions A beneficial effect of PHBC treatment is suggested because the proportion of responders was significantly higher in this group of patients.

The Prevalence of MRI-Defined Sacroiliitis and Classification of Spondyloarthritis in Patients with Acute Anterior Uveitis: A Longitudinal Single-Centre Cohort Study.

Background: Acute anterior uveitis (AAU) is a relatively common extra-musculoskeletal manifestation of axial spondyloarthritis (axSpA); however, data on the prevalence of active sacroiliitis in patients with AAU are limited. Methods: 102 patients with AAU and 39 healthy subjects (HS) underwent clinical assessment and sacroiliac joint MRI. Patients with absence of active sacroiliitis were reassessed after two years. International Spondyloarthritis Society (ASAS) classification criteria for axSpA (regardless of patient’s age) and expert opinion for definitive diagnosis of axSpA were applied. Results: Although chronic back pain was equally present in both groups, bone marrow edema (BME) in SIJ and BME highly suggestive of axSpA was found in 52 (51%) and in 33 (32%) patients with AAU compared with 11 (28%) and none in HS, respectively. Out of all AAU patients, 41 (40%) patients fulfilled the ASAS classification criteria for axSpA, and 29 (28%) patients were considered highly suggestive of axSpA based on clinical features. Two out of the 55 sacroiliitis-negative patients developed active sacroiliitis at the two-year follow-up. Conclusions: One-third of patients with AAU had active inflammation on SIJ MRI and clinical diagnosis of axSpA. Therefore, patients with AAU, especially those with chronic back pain, should be referred to a rheumatologist, and the examination should be repeated if a new feature of SpA appears.

Evaluation of corneal endothelial cell morphology by specular microscopy in patients with acute anterior uveitis.

To determine the structure and morphology of corneal endothelial cell layer in patients with acute anterior uveitis. Thirty-four eyes of 34 acute anterior uveitis patients and 34 eyes of 34 healthy subjects were included. Mean cell density, coefficient of variation, maximum cell area, minimum cell area, average cell area and hexagonality ratio values were evaluated by non-contact specular microscopy. Parameters recorded in both groups were compared. The mean maximum cell area was 1054,44 ± 251,14 µm2, minimum cell area was 152.29 ± 53.65 µm2 and average cell area was 386.91 ± 41.73 µm2 in acute anterior uveitis group and the mean maximum cell area was 1057.65 ± 261.23 µm2, minimum cell area was 147.26 ± 20.45 µm2 and average cell area was 383.53 ± 43.12 µm2 in the control group. There were no significant differences between the two groups in terms of maximum, minimum and average cell area (respectively, p = 0.080, p = 0.72, p = 0.62, p = 0.67). The mean cell density was 2607.74 ± 277.63 cells/ µm2 in acute anterior uveitis group and 2669.35 ± 265.22 cells/µm2 in the control group. (p = 0.358). In acute anterior uveitis group the mean coefficient of variation was 31.68 ± 8.16, hexagonality ratio was 63.85 ± 11.14 and mean central corneal thickness was 571.47 ± 55.99µm; in control group the mean coefficient of variation was 25.29 ± 3.00, mean hexagonality ratio was 72.6 ± 4.80% and mean central corneal thickness was 534.82 ± 33.84µm. Statistically significant differences were seen between the two groups (respectively, P = 0,00, P = 0,00, P = 0.02). The mean central corneal thickness and coefficient of variation values were found higher, and the hexagonality ratio was found lower in acute anterior uveitis group. Our findings suggest that intraocular inflammation in anterior chamber negatively affects the endothelial function in patients with acute anterior uveitis.

The acute phase response protein SERPINA3 is increased in tear fluid from the unaffected eyes of patients with unilateral acute anterior uveitis

BackgroundTo identify candidate tear fluid biomarkers in patients with unilateral acute anterior uveitis (AAU) that can aid in the differentiation between these patients and patients with bacterial keratitis or healthy controls.MethodsThirteen patients (40.1 ± 16.2 years of age) with unilateral AAU, seven patients with unilateral bacterial keratitis (40.2 ± 15.3 years of age), and 14 healthy subjects (41.1 ± 11.6 years of age) were included. The tear proteome of affected eyes was compared with that of the unaffected eye or healthy controls. Proteins were identified by liquid chromatography tandem mass spectrometry and enzyme-linked immunosorbent assay.ResultsRelative protein ratios were detected and calculated for 272 unique proteins. Compared with healthy controls and the unaffected eye, the top upregulated proteins in AAU eyes were submaxillary gland androgen regulated protein 3B (SMR3B) and SMR3A. Similarly, the top upregulated proteins in bacterial keratitis were S100 calcium-binding protein A9 and orosomucoid 2. The acute phase response protein Serpin Family A Member 3 (SERPINA3) was increased in the healthy eye of AAU patients (P = 0.019) compared with healthy controls. Laser flare measurements in affected eyes of AAU patients showed positive logarithmic correlation with SERPINA3 in tear samples of the unaffected eye (P = 0.022). The use of SERPINA3 as a tear biomarker yielded a sensitivity of 85% and a specificity of 71% in detecting patients with AAU in the study population.ConclusionsThe acute phase response protein SERPINA3 was increased in tear samples of unaffected eyes of patients with unilateral AAU compared with healthy controls. This study highlights SERPINA3 as a potential biomarker for AAU. Future research should explore the dynamic properties of SERPINA3 in the tear fluid of active and quiescent uveitis eyes.

POS0410 BIOMARKERS REFLECTING DISTURBED GUT BARRIER DIFFER IN PATIENTS WITH SPONDYLOARTHRITIS, CROHN’S DISEASE AND ACUTE ANTERIOR UVEITIS

Background:Spondyloarthritides (SpA) are characterized by frequent extra-musculoskeletal manifestations (EMM) among them acute anterior uveitis (AAU) and Crohn’s disease (CD). Vice versa, about 50% of AAU and 20% of CD patients have concomitant SpA. SpA patients show gut dysbiosis together with frequent subclinical gut inflammation. Biomarkers reflecting disturbed gut barrier (intestinal-fatty acid binding protein (iFABP), lipopolysaccharide binding protein (LBP) and zonulin) were previously found to be elevated in patients with radiographic axial SpA (r-axSpA) (1).Objectives:To evaluate whether biomarkers reflecting leaky gut are altered in patients with AAU, CD and SpA compared to healthy controls and whether they differ between patients with EMM with and without concomitant SpA.Methods:A total of 100 patients from the German Spondyloarthritis Inception Cohort (GESPIC) were included – among them 20 patients with r-axSpA without EMM, 40 patients with CD and 40 patients with non-infectious AAU – out of which 19 and 20 patients, respectively, had concomitant SpA (11/8 and 20/0 axial/peripheral SpA, respectively). The GESPIC patients were compared to 20 age- and sex-matched healthy donors (HD). The following five serum biomarkers were analyzed with ELISA: calprotectin, iFABP, LBP, soluble CD14 (sCD14) and zonulin.Results:Patient characteristics are shown in Table 1. Serum levels of calprotectin, LBP, sCD14 and zonulin differed significantly between patients with r-axSpA, AAU and CD with and without concomitant SpA and HD (Figure 1). When comparing patients with EMM with and without underlying SpA, calprotectin serum levels were significantly elevated in CD patients with SpA (8.6µg/ml (SD 5.5µg/ml)) compared to CD patients without SpA (5.7µg/ml (SD 4.1µg/ml); Mann-Whitney U Test, p=0.031). Serum levels of the analyzed biomarkers did not differ between AAU patients with and without axSpA. Spearman rank correlation revealed a significant association between CRP and calprotectin (correlation coefficient r=0.230; p=0.012), LBP (r=0.596; p&lt;0.0001), sCD14 (r=0.428; p&lt;0.0001) and zonulin (r=0.221; p=0.016), respectively. Furthermore, LBP and zonulin serum levels correlated positively (r=0.208; p=0.023); as well as LBP and sCD14 levels (r=0.418; p&lt;0.0001).Table 1.Patient characteristics. Mean values (standard deviation) or absolute numbers are shown.CD + SpACDr-axSpAAAU + axSpAAAUHDN192120202020Age39.1 (11.3)38.7 (14.4)38.4 (10.3)39.6 (12.0)39.2 (12.5)38.6 (12.9)Male (%)9 (47%)9 (43%)9 (45%)9 (45%)9 (45%)9 (45%)HLA-B27 positive (%)5 (26%)3 (14%)17 (85%)17 (85%)13 (65%)2 (10%)CRP in mg/l14.3 (25.6)18.0 (41.8)9.1 (11.3)6.7 (9.9)2.3 (3.4)0.6 (0.7)ASDAS2.8 (1.1)3.2 (0.6)2.2 (1.0)BASDAI4.1 (2.2)5.4 (1.2)3.2 (2.4)Figure 1.Biomarkers reflecting disturbed gut barrier show distinct signatures in patients with acute anterior uveitis, Crohn’s disease and axial Spondyloarthritis. Kruskal Wallis Test; p values shown. Dunn-Bonferroni Post-Hoc analyses, significant pairwise differences are marked; * p&lt;0.05; **p&lt;0.01; ***p&lt;0.0001Conclusion:We found substantial differences in biomarkers reflecting disturbed gut barrier between with SpA, CD, AAU and healthy controls. The presence of SpA was associated with higher calprotectin serum levels in CD as compared to CD without SpA.

Flow cytometric analysis of T lymphocytes and cytokines in aqueous humor of patients with varicella zoster virus-mediated acute retinal necrosis

BackgroundThe purpose of this study is to investigate the aqueous humor (AH) T lymphocyte subsets and cytokines of acute retinal necrosis (ARN) to elucidate the immunologic inflammatory features of this disorder.MethodsThree patients with ARN infected with varicella zoster virus (VZV) who underwent multiple intravitreal injections of ganciclovir were enrolled in this study. The control group consisted of four non-infectious patients with acute anterior uveitis (AAU). Flow cytometric analysis was performed on the lymphocyte subsets from the AH and peripheral blood (PB) samples during the active phase of intraocular inflammation. Five inflammatory cytokines were measured in each AH sample and various clinical characteristics were also assessed.ResultsVZV deoxyribonucleic acid (DNA) was detected by real-time polymerase chain reaction (PCR) in AH from all the ARN patients, who showed higher CD8+ T lymphocytes population in AH than the AAU patients (p = 0.006). CD4/CD8 ratios of T lymphocytes and the percentage of CD8 + CD25+ T lymphocytes in AH were significantly lower in ARN than in AAU (p = 0.006; p = 0.012). In the ARN patients, the percentages of CD4+ and CD8+ T lymphocytes in AH were higher than those found in PB. The percentage of CD4 + CD25+ T lymphocytes in AH was significantly higher than the proportion in PB in the AAU patients (p = 0.001). Immunoregulatory cytokine Interleukin-10 in AH was significantly elevated in the ARN patients in comparison with the case of the AAU patients (p = 0.036). In ARN, the copy number of VZV DNA in AH positively correlated with the percentage of CD8+ T lymphocytes in AH and negatively correlated with the CD4/CD8 ratio in AH during the course of disease treatment (p = 0.009, r = 0.92; p = 0.039, r = − 0.834).ConclusionThe ARN patients caused by VZV had different intraocular T lymphocyte subsets and cytokines profile than those of the non-infectious patients. High percentages of CD8+ T lymphocytes and low CD4/CD8 T cell ratios may be a potential biomarker for diagnosis of viral-infectious uveitis. T lymphocytes examination at the inflammatory sites has the potential to become a useful research tool for differentiating viral and non-viral uveitis.

Mesalazine Suppresses Proinflammatory Cytokines in Patients with Acute Anterior Uveitis Independently of HLA-B27

ABSTRACT Purpose: The aim is to unravel the mechanism of mesalazine (5-ASA) on proinflammatory cytokines in PBMCs of patients with HLA-B27 +and HLA-B27 -acute anterior uveitis (AAU), and whether this may explain the different effects of 5-ASA in both disoders. Methods: PBMCs from 12 HLA-B27+ and 4 HLA-B27- AAU patients were preincubated with 5-ASA and stimulated with LPS. As mesalazine (5-ASA) could be involved in ER stress, proinflammatory and ER stress-associated cytokines and markers were measured. Results: Mesalazine (5-ASA) suppressed IL-6 mRNA in healthy donors and in HLA-B27+ and HLA-B27- patients but did not lead to induction and secretion of IL-1β. In HLA-B27 + or – patients the ER stress-associated markers CHOP (DDIT3) and ATF6 were suppressed. Conclusions: Here we show that mesalazine (5-ASA) inhibits the transcription of proinflammatory and (ER) stress associated cytokines and markers, independently of the HLA-B27 status. Results show the similarities of both AAU types but do not decipher the mechanism why the HLA-B27 status determines the therapeutic response to mesalazine in AAU.

Linoleic acid inhibits in vitro function of human and murine dendritic cells, CD4+T cells and retinal pigment epithelial cells.

Increased linoleic acid (LA) was observed in acute anterior uveitis (AAU) patient feces in our previous study. To investigate the immunoregulatory effect of LA, we studied the effect of LA on human and murine dendritic cells (DCs), CD4+T cells, and retinal pigment epithelial (RPE) cells in vitro. The level of LA in feces from AAU patients and healthy individuals was measured by gas chromatography coupled with a mass spectrometer (GC-MS). The immunoregulatory effect of LA on human and murine DCs, CD4+ T cells, and RPE cells was evaluated by enzyme linked immunosorbent assay (ELISA) and flow cytometry (FCM). The effect of LA on DCs was evaluated by Tandem mass tag (TMT)-based proteomics analysis. Increased LA was observed in feces from AAU patients (1018.35 ± 900.01 mg/kg) as compared with healthy individuals (472.55 ± 365.49 mg/kg, p = 0.0136). LA attenuated the antigen-presenting function of human and murine DCs by decreasing the expression of CD40, the secretion of IL-6 and IL-12p70, and the ability to shift naïve T cells towards T helper type 1 (Th1) and Th17 cells. LA also inhibited the secretion of MCP-1 and IL-8 from RPE cells. Proteomics analysis showed differential expression of 28 proteins, including squalene epoxidase (SQLE), farnesyl-diphosphate farnesyltransferase 1 (FDFT1), and cytochrome P450 family 51 subfamily A member 1 (CYP51A1), in LA-treated DCs compared with controls. LA also accelerated the apoptosis of DCs from healthy individuals. LA inhibited the function of human and murine DCs, CD4+T cells, and RPE cells, regulated the expression of proteins, and promoted the apoptosis of human DCs. These results collectively suggest that LA might decrease the function of immune cells in vitro, and further studies are needed to investigate its role in the pathogenesis of AAU.

Clinical Features of Acute Anterior Uveitis with Complicated Course in a Slovenian Patient Population.

Purpose To report on patients who needed hospitalization due to acute anterior uveitis (AAU) and within this group to compare clinical features and outcomes of treatment of HLA-B27+ and HLA-B27− AAU in the population of Slovenian patients. Methods Retrospective study of hospitalized patients with AAU in the last 39 months at the Eye Hospital in Ljubljana. The data of AAU patients were retroactively studied and compared on the basis of HLA-B27 antigen presence: visual acuity upon admission, visual outcome, the presence of hypopyon, fibrinous reaction, posterior iris synechiae, and complications, such as elevated intraocular pressure, cataract, and cystoid macular edema (CME). We compared the investigations in the diagnostic process, the associated systemic disease, and the treatment administered. Statistical analyses included Student's t-test Fisher's exact test, and the Kolmogorov–Smirnov test. A p value of <0.05 was considered statistically significant. Results A total of 37 hospitalized patients with AAU were included. HLA-B27 antigen was detected in 73% of patients. In the HLA-B27+ group, women were more commonly affected, while the males were more affected in the HLA-B27− group. The occurrence of fibrin was significantly more common in HLA-B27+ patients, as well as hypopyon and posterior synechiae; only fibrin reached the statistical significance (p < 0.05). The incidence of cataracts, ocular hypertension, and glaucoma did not differ significantly between the two groups. HLA-B27+ AAU was more often associated with systemic diseases, and patients in this group were more frequently treated with systemic immunomodulatory drugs, however, no difference reached the statistical significance. We did not notice any major differences in the final visual acuity in the comparing groups. Conclusion Almost ¾ of AAU patients that required hospitalization were HLA-B27+. In this group, disease was more severe, more frequently associated with ocular complications and systemic disease, but final visual acuity was the same in both groups. HLA-B27 typing has no prognostic value in our group of complicated AAU patients, but it eases the decision about necessary diagnostics and treatment.

Analysis of the role of palmitoleic acid in acute anterior uveitis

PurposeTo study the role of palmitoleic acid (PA) in the pathogenesis of acute anterior uveitis (AAU). MethodsPA levels in feces from AAU patients were measured by gas chromatography coupled with a mass spectrometer (GC-MS) and compared with samples obtained from healthy individuals. Enzyme linked immunosorbent assay (ELISA) and flow cytometry (FCM) were used to assess the effect of PA on dendritic cells (DCs) and CD4+T cells obtained from mice, AAU patients and healthy individuals. C57BL/6 mice were fed with PA or vehicle and experimental autoimmune uveitis (EAU) was induced with a human retinal IRBP651-670 peptide. Disease severity of EAU was evaluated by clinical manifestation and histology. Differentiation of splenic Type 1 helper T cells (Th1) and Th17 cells was evaluated by FCM. Tandem mass tag (TMT)-based proteomics analysis was used to identify differentially expressed proteins following incubation of DCs with PA. ResultsThe fecal concentration of PA was increased in AAU patients as compared with healthy individuals. In vitro, PA promoted apoptosis of DCs and inhibited the secretion of TNF-α from mouse bone-marrow-derived dendritic cells (BMDCs) as well as in DCs from AAU patients and healthy individuals. It only decreased DCs surface marker expression and IL-12p70 secretion in BMDCs and healthy individuals DCs but not in AAU patient DCs. PA-treated BMDCs inhibited Th cell differentiation from mouse naïve CD4+T cells and IL-17 and IFN-γ secretion in co-culture supernatants. PA also inhibited the differentiation of Th cells and secretion of IFN-γ and IL-17 in CD4+T cells from mice, AAU patients and healthy individuals. In vivo, PA-treated EAU mice showed milder clinical and histopathological intraocular manifestations as compared with the control group. PA feeding inhibited differentiation of splenic Th17 cells, whereas Th1 cells were not affected. Up to 30 upregulated and 77 downregulated proteins were identified when comparing PA-treated DCs with controls. ConclusionAn increased expression of fecal PA was observed in AAU patients. PA was shown to have immunoregulatory effects on DCs and CD4+T cells and attenuated disease severity in EAU mice.

Uveitis in spondyloarthritis.

Uveitis is the most frequent extra-articular manifestation of axial spondyloarthritis (SpA), occurring in up to one-third of the patients. In the majority of patients, uveitis is acute, anterior and unilateral and presents with photosensitivity, sudden onset of pain and blurred vision. Topical steroids are an effective treatment; however, recurrent or refractory cases may need conventional disease-modifying antirheumatic drugs or biological treatment with monoclonal tumor necrosis factor (TNF) inhibitors, thus also influencing treatment strategy of the underlying SpA.Though the exact pathogenesis of SpA and uveitis remains unknown, both seem to result from the interaction of a specific, mostly shared genetical background (among other HLA-B27 positivity), external influences such as microbiome, bacterial infection or mechanical stress and activation of the immune system resulting in inflammation. Up to 40% of patients presenting with acute anterior uveitis (AAU) have an undiagnosed SpA. Therefore, an effective referral strategy for AAU patients is needed to shorten the diagnostic delay of SpA and enable an early effective treatment. Further, the risk for ophthalmological manifestations increases with the disease duration in SpA; and patients presenting with ocular symptoms should be referred to an ophthalmologist. Thus, a close collaboration between patient, rheumatologist and ophthalmologist is needed to optimally manage ocular inflammation in SpA.

Correlation of serum amyloid A levels, clinical manifestations, treatment, and disease activity in patients with acute anterior uveitis.

PurposeTo investigate the association between serum amyloid A (SAA) protein and the clinical features of acute anterior uveitis (AAU), and to evaluate the disease activity and treatment effect in relation to SAA levels.MethodsAAU patients and healthy individuals were recruited from October 2016 to August 2017 at the Department of Uveitis, in the Eye Hospital of Wenzhou Medical University. Related demographic, clinical characteristics, and therapeutic data were analyzed.ResultsOne hundred and eight AAU patients and 18 healthy controls were included in this study. Serum SAA levels in AAU patients were significantly higher than those of healthy controls (p all < 0.0001). Significantly higher SAA levels were found in AS+AAU patients than those in AS−AAU patients (p < 0.05). SAA levels were also significantly higher in patients with HLA-B27+AAU compared with those with HLA-B27−AAU (p < 0.05). Furthermore, in each of the AAU subgroups, higher SAA levels were observed in the active state than those in the inactive state (p all < 0.05). In addition, SAA levels were positively correlated to anterior chamber cell counts (r = 0.492, p < 0.0001). ROC curve analysis revealed that SAA had an AUC value of 0.727 for detecting active inflammation (Youden’s index = 0.38). SAA decreased with effective treatments (p = 0.0002).ConclusionSerum levels of SAA were elevated in AAU patients. The increased levels of SAA were correlated with AS and HLA-B27 status. SAA levels were also positively correlated to disease activity and decreased with effective treatments. These findings suggest that SAA is associated with AAU, with a potential role in monitoring inflammatory processes and assessing the efficacy of therapy.

Tear proteome in uveitis

AbstractPurposeUveitis represents the fourth commonest cause of legal blindness in adult patients. Analysis of tear fluid proteins in acute anterior uveitis (AAU) patients could represent a novel diagnostic modality. This study aimed at detecting potential tear fluid biomarkers in patients with AAU and bacterial keratitis (BK).MethodsFourteen patients with unilateral AAU, seven patients with bacterial keratitis (BK) and 14 healthy age‐ and sex‐matched control subjects were included. Tear fluid samples were obtained by inserting two Schirmer’s test strips behind the lower eyelid of each eye for five minutes without anesthesia. Proteins were then identified by liquid chromatography tandem mass spectrometry. Pooled tear fluid samples from healthy controls were compared with samples from AAU and BK patients, respectively. Ingenuity Pathway Analysis (IPA) was used to explore biological function.ResultsA relative protein level was obtained for 235 proteins in AAU patients and 198 proteins in BK patients, while using healthy control subjects as reference. In AAU patients, nine proteins were at least two‐fold up‐regulated, whereas in BK patients, 12 proteins were up‐regulated more than two‐fold. None of the top ten upregulated proteins in AAU and BK overlapped. The top canonical pathway identified by IPA was ‘Acute Phase Response Signaling’ in AAU and ‘LXR/RXR activation’ in BK. Top upstream activated regulators included peroxisome proliferator activated receptor delta in AAU and interleukin 6 in BK.ConclusionsTear fluid protein analysis represents a promising diagnostic modality for ocular inflammatory diseases and demonstrate differentially expressed proteins in acute anterior uveitis and bacterial keratitis.