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1.
Invest Ophthalmol Vis Sci ; 61(10): 5, 2020 08 03.
Article in English | MEDLINE | ID: mdl-32749463

ABSTRACT

Purpose: To examine the role of aqueous tumor necrosis factor α (TNF-α)-RhoA-Rho kinase (ROCK) signaling in cytomegalovirus (CMV)-induced apoptosis and the barrier function of cultured human corneal endothelial cells (hCECs) in CMV-positive Posner-Schlossman syndrome (CMV+/PSS) patients. Methods: Aqueous levels of TNF-α, IL-8, IL-10, and several other cytokines in 19 CMV+/PSS patients and 20 healthy control subjects were quantitated using a multiplex assay. The expression of active RhoA in hCECs post-CMV infection was determined using western blotting (WB). The expression levels of TNF-α and nuclear factor kappa B (NF-κB) in CMV-infected hCECs were examined by immunocytochemistry (ICC) and WB with and without ROCK inhibitors. The apoptotic rate and barrier integrity in CMV-infected hCECs were also examined. Results: The expression levels of TNF-α, monocyte chemoattractant protein-1 (MCP-1), IL-8, and IL-10 were upregulated in the aqueous humor of CMV+/PSS patients, and among these upregulated cytokines aqueous TNF-α was negatively correlated with the number of corneal endothelial cells. In CMV-infected hCECs, upregulation of TNF-α and NF-κB was determined by WB and ICC. In hCECs, CMV infection induced apoptosis and significantly impaired cell-cell contacts, effects that were attenuated by treatment with a ROCK inhibitor. Conclusions: Aqueous TNF-α was upregulated in CMV+/PSS patients, which may have triggered corneal endothelial cell loss. Modulation of TNF-α, including its downstream Rho-ROCK signaling, could serve as a novel treatment modality for corneal endothelial cell loss in CMV+/PSS patients.


Subject(s)
Apoptosis/drug effects , Cytomegalovirus Infections/pathology , Endothelium, Corneal/pathology , Enzyme Inhibitors/pharmacology , Eye Infections, Viral/pathology , Iridocyclitis/pathology , rho-Associated Kinases/antagonists & inhibitors , Adult , Aged , Aged, 80 and over , Amides/pharmacology , Aqueous Humor/metabolism , Aqueous Humor/virology , Blotting, Western , Cells, Cultured , Cytokines/metabolism , Cytomegalovirus/isolation & purification , Cytomegalovirus Infections/metabolism , Cytomegalovirus Infections/virology , Endothelium, Corneal/metabolism , Endothelium, Corneal/virology , Eye Infections, Viral/metabolism , Eye Infections, Viral/virology , Female , Humans , Immunohistochemistry , Iridocyclitis/metabolism , Iridocyclitis/virology , Isoquinolines/pharmacology , Male , Middle Aged , Multiplex Polymerase Chain Reaction , Prospective Studies , Pyridines/pharmacology , Sulfonamides/pharmacology , rhoA GTP-Binding Protein/metabolism
2.
PLoS One ; 7(9): e44453, 2012.
Article in English | MEDLINE | ID: mdl-23028541

ABSTRACT

AIM: To study the differences in aqueous cytokines in Posner-Schlossman Syndrome (PSS) patients with and without human cytomegalovirus (CMV) DNA in the aqueous humor. METHODS: This is a prospective study. Fifty-three uveitis patients with clinical signs of PSS were enrolled and aqueous humor samples were collected. Fourteen PSS patients were positive of CMV DNA in the aqueous by polymerase chain reaction (PCR) analysis. These eyes were negative of common ocular pathogens such as herpes simplex virus, varicella-zoster virus, rubella virus and toxoplasma. Twenty-five otherwise healthy cataract patients were enrolled as controls. Cytokine concentration was measured by a magnetic color-bead-based multiplex assay and analyzed using statistical and classification approaches. RESULTS: The average age of 53 PSS patients was 48.74±13.43 years (yrs) (mean ± standard deviation) and 66.3±15.0 yrs for the controls. The median CMV viral DNA copy number was 26000/mL aqueous (range 1400 to 85000 copies/mL) in 14 CMV positive patients as determined by quantitative PCR. PSS aqueous had significantly higher Interleukin (IL)-8 (CXCL8), monocyte chemotactic protein-1 (CCL2), macrophage inhibitory protein 1-ß (CCL4), granulocyte colony-stimulating factor (GCSF) and transforming growth factor-ß (TGF-ß) levels than controls after adjusted by age and gender. IL-2, IL-12, tumor necrosis factor-α (TNF-α) and interferon-α (IFN-α) levels were significantly lower in PSS aqueous than controls. No difference between CMV positive PSS and CMV negative PSS aqueous was observed. Over 97% of PSS samples were distinguished from controls by elevated CXCL10 (>500 ng/mL), CXCL8 (>30 ng/mL) and CCL2 (>60 ng/mL) levels. CONCLUSION: PSS eyes were characterized by elevated aqueous chemokine concentration. The presence of CMV viral DNA was not associated with significant change of the type of cytokine expression in PSS patients.


Subject(s)
Cytokines/metabolism , Uveitis/metabolism , Adult , Aqueous Humor/metabolism , Cytomegalovirus Infections/metabolism , Female , Glaucoma/metabolism , Glaucoma/virology , Humans , Iridocyclitis/metabolism , Iridocyclitis/virology , Male , Middle Aged , Polymerase Chain Reaction , Prospective Studies , Uveitis/virology
4.
Invest Ophthalmol Vis Sci ; 42(2): 328-32, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11157862

ABSTRACT

PURPOSE: Increased production of 12-hydroxyeicosatetraenoic acid [12(R)-HETE] and 12-hydroxyeicosatrienoic acid [12(R)-HETrE] positively correlates with the in vivo progression of ocular surface inflammation in rabbits. Tear film was collected from human subjects with inflamed eyes to determine whether these eicosanoids could be detected from endogenous sources. METHODS: Control and inflamed eyes were assessed and assigned a subjective inflammatory score. Tears were collected and extracted with an internal standard. Single-ion-monitoring gas chromatography-mass spectrometry (SIM-GC-MS) was performed to quantitate endogenous levels of 12-HETE and 12-HETrE. RESULTS: 12-HETrE was detected in the tear film of both control and inflamed eyes, with the mean level being seven times higher in inflamed tears. 12-HETE was not detected in control tears and was detected in only 6 of 38 inflamed-eye tear samples. CONCLUSIONS: The current findings demonstrate that the human eye produces detectable amounts of 12-HETrE, which is released into the tear flow. The increased levels of 12-HETrE associated with ocular surface inflammation suggest that this eicosanoid may contribute to inflammation of the ocular surface in humans.


Subject(s)
12-Hydroxy-5,8,10,14-eicosatetraenoic Acid/metabolism , Tears/metabolism , Arachidonic Acid/metabolism , Conjunctivitis/metabolism , Eye Foreign Bodies/metabolism , Gas Chromatography-Mass Spectrometry , Humans , Inflammation/metabolism , Iridocyclitis/metabolism , Keratitis/metabolism , Keratitis, Herpetic/metabolism , Keratoconjunctivitis/metabolism
5.
Acta Ophthalmol Scand ; 78(5): 516-8, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11037905

ABSTRACT

PURPOSE: To investigate the influence of protein concentration in the anterior chamber, measured by laser flare meter, on pain sensation after phenylephrine instillation in patients with iridocyclitis. METHODS: Twenty-five consecutive patients with iridocyclitis were included. Patients with cataract, exfoliation syndrome, diabetes mellitus, glaucoma or any other previous ocular diseases or ocular surgery were excluded. Patients were divided into two groups: Group 1--without fibrinoid reaction (FR) in the anterior chamber (18 patients), and Group 2--with FR (7 patients). Protein concentration in the anterior chamber was measured with laser flare meter (FC 500, Kowa Co., Japan). Pupil size was measured by Alcon Tilo Scale, and pain sensation was estimated by Visual Analogue Scale (VAS, Kabi Pharmacia). All measurements were done before and 1 hour after topical instillation of 10% phenylephrine hydrochloride into the subconjunctival sac of the inflamed eyes. RESULTS: Eyes with iridocyclitis and fibrinoid reaction (FR) have a higher flare intensity compared to those without FR (p<0.05). Pupil size was significantly increased after phenylephrine instillation in both study groups (Wilcoxon test, p<0.05). The VAS pain and flare intensity were significantly decreased in group without FR after phenylephrine instillation (Group 1) compared to values before treatment (Wilcoxon test, p<0.05). In eyes with FR (Group 2), no significant influence of phenylephrine instillation was found on VAS pain and flare intensity. CONCLUSIONS: After phenylephrine instillation, flare intensity and pain were significantly decreased only in eyes with iridocyclitis and without FR. The decreasing level of flare intensity, and paralysis of the pupil after phenylephrine instillation seem to alleviate pain in those eyes.


Subject(s)
Anterior Chamber/metabolism , Eye Proteins/metabolism , Iridocyclitis/drug therapy , Mydriatics/therapeutic use , Pain/prevention & control , Phenylephrine/therapeutic use , Pupil/drug effects , Adult , Aged , Aged, 80 and over , Aqueous Humor/metabolism , Diagnostic Techniques, Ophthalmological , Female , Humans , Iridocyclitis/metabolism , Male , Middle Aged , Mydriatics/administration & dosage , Ophthalmic Solutions , Pain/metabolism , Pain Measurement , Phenylephrine/administration & dosage , Prospective Studies
6.
Br J Ophthalmol ; 83(6): 703-6, 1999 Jun.
Article in English | MEDLINE | ID: mdl-10340980

ABSTRACT

AIMS: To analyse the unusual physical sign of iris crystals occurring in patients with uveitis. METHODS: Demographic details and clinical features were documented in 24 patients with chronic uveitis and iris crystals. Plasma immunoglobulin subclasses were measured, and a histopathological review of iridectomy specimens from 33 patients with chronic uveitis was also undertaken. RESULTS: The mean age of patients was 38 years, with a slight preponderance of females. 17 patients had Fuchs' heterochromic cyclitis although a number of other uveitis entities were represented. There was no correlation between severity of clinical signs and presence of iris crystals. Over a mean follow up period of 15 months no significant change in the number, size, or position of the crystals was seen except in four patients who underwent intraocular surgery. Only three patients had raised plasma IgG1. The review of the histology of iridectomy specimens failed to show evidence of Russell body formation in any patient. CONCLUSIONS: Iris crystals appear to be rare but may be underreported as they are small and can easily be missed. They are likely to be associated with disease processes in which there is active immunoglobulin production within the anterior chamber, such as Fuchs' heterochromic cyclitis.


Subject(s)
Iridocyclitis/metabolism , Iris/metabolism , Uveitis/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Chronic Disease , Crystallization , Female , Follow-Up Studies , Humans , Male , Middle Aged
8.
Acta Ophthalmol Scand ; 76(4): 405-12, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9716325

ABSTRACT

PURPOSE: To study aqueous flare longitudinally in patients with Fuchs' heterochromic uveitis (FHU) and for comparison in another group with iris atrophy (non FHU). METHODS: A new laser flare meter was used. Iris angiography was performed in most FHU patients. RESULTS: The flare values in the affected eyes in the FHU group were always higher compared to the normal eyes and rather constant over time. Cataract surgery did not permanently change the flare values. Systemic - but not local steroid therapy caused normalization of the flare. Iris angiography displayed leakage in all FHU eyes. Leakage from original vessels dominated although some newly-formed leaking vessels were observed. CONCLUSIONS: The results support the notion that FHU is a low grade, chronic, stable disease. Leaking iris vessels, original or newly-formed, are probably the source of the increased flare. Our hypothesis is that inflammation plays a major part. Atrophy per se does not enhance flare.


Subject(s)
Aqueous Humor/metabolism , Fluorescein Angiography , Iridocyclitis/metabolism , Iris/blood supply , Iris/pathology , Adult , Aged , Atrophy/metabolism , Atrophy/pathology , Capillary Permeability , Cataract Extraction , Chronic Disease , Female , Fluorophotometry/instrumentation , Fluorophotometry/methods , Humans , Inflammation/metabolism , Inflammation/pathology , Iridocyclitis/pathology , Male , Middle Aged
9.
Curr Eye Res ; 13(8): 603-10, 1994 Aug.
Article in English | MEDLINE | ID: mdl-7956313

ABSTRACT

The purpose of this study was to investigate the effect of a novel immunomodulator, AS101 [ammonium trichloro(dioxyethelene-O-O') tellurate], in the eye. Lewis rats were injected intravitreally with AS101 at a concentration of 13 micrograms/ml in one eye and BSS in the contralateral eye. Control animals were injected with BSS into the central vitreous of both eyes. Ocular inflammation was evaluated at 20 hours by histology, immunopathology, and by cell count, protein and cytokine measurement in the aqueous humor. At 20 hours, eyes injected with AS101 developed iridocyclitis and mild vitritis versus minimal inflammation and/or protein in contralateral eyes or eyes of control animals (p = 0.0121). The inflammatory infiltrate was mixed in character. Major Histocompatibility Complex (MHC) class II antigens and intercellular adhesion molecules (ICAM-1) were expressed in the anterior segment of eyes injected with AS101. In the aqueous humor of these eyes there were significant quantities of inflammatory cells, protein (mean +/- SEM = 11.2 +/- 2.3 mg/ml) and the cytokine interleukin 6 (IL-6) (450 units/ml) compared with contralateral eyes (p = 0.0005 for inflammatory cells; protein, mean +/- SEM = 1.6 +/- 0.17 mg/ml; IL-6 = 12 units/ml) and both eyes of control animals injected with BSS (p = 0.8955 for inflammatory cells; protein, OD = 1.5 mg/ml, OS = 0.7 mg/ml; IL-6, OD = 8 units/ml, OS = 13 units/ml). AS101 has a local inflammatory effect in the eye. This compound may activate ocular inflammation by releasing cytokines such as IL-6.


Subject(s)
Adjuvants, Immunologic/pharmacology , Ethylenes/pharmacology , Iridocyclitis/chemically induced , Vitreous Body/drug effects , Animals , Anterior Eye Segment/drug effects , Anterior Eye Segment/metabolism , Anterior Eye Segment/pathology , Eye Diseases/chemically induced , Eye Diseases/metabolism , Eye Diseases/pathology , Female , Histocompatibility Antigens Class II/metabolism , Inflammation , Injections , Intercellular Adhesion Molecule-1/metabolism , Interleukin-6/metabolism , Iridocyclitis/metabolism , Iridocyclitis/pathology , Rats , Rats, Inbred Lew , Vitreous Body/metabolism , Vitreous Body/pathology
10.
Br J Ophthalmol ; 78(7): 593, 1994 Jul.
Article in English | MEDLINE | ID: mdl-7918281
11.
Invest Ophthalmol Vis Sci ; 33(10): 2878-84, 1992 Sep.
Article in English | MEDLINE | ID: mdl-1526738

ABSTRACT

The laser flare cell meter quantifies anterior chamber (AC) protein (flare) by measuring light scattering of a helium-neon laser beam in the AC. The relationship between photon count and protein concentration both in vitro and in vivo was assessed. The reproducibility of the in vitro photon count measurements was 7.3%. There was a significant linear relationship between photon count and the concentration of both albumin (r = 1.0, P = 0.0001) and immunoglobulin G (IgG, r = 0.99, P = 0.0001) in vitro, but the linear-regression formulas were different with greater light scattering by IgG than by albumin at the same concentration. Laser flare measurements were done on 22 patients (12 normal eyes, 5 eyes with Fuchs' heterochromic cyclitis, and 5 uveitic eyes) before cataract surgery. Aqueous humor obtained from these patients by paracentesis was analyzed for total protein, albumin, and IgG concentration. There was a significant linear relationship (r = 0.88, P = 0.0001) between the laser flare value (range, 5.8-107.8 photons/msec) and the total aqueous protein concentration (range, 14-388 mg/dl). Use of an in vitro albumin calibration curve to convert photon count into protein concentration was found to overestimate the actual protein concentration. This overestimation was slight in normal eyes and increased with increased blood-aqueous barrier breakdown. The use of such a calibration curve therefore is not appropriate in studies on diseased eyes. The authors recommended that laser flare results be expressed in either photons per milliseconds or converted into an equivalent protein concentration using a calibration curve based on actual AC protein measurements.


Subject(s)
Anterior Chamber/metabolism , Eye Proteins/analysis , Lasers , Aged , Aged, 80 and over , Albumins/analysis , Aqueous Humor/metabolism , Biological Transport, Active , Blood/metabolism , Cataract Extraction , Chronic Disease , Humans , Immunoglobulin G/analysis , Iridocyclitis/metabolism , Middle Aged , Reproducibility of Results , Uveitis/metabolism
12.
Int Ophthalmol ; 16(1): 15-22, 1992 Jan.
Article in English | MEDLINE | ID: mdl-1537644

ABSTRACT

Aqueous humour from patients with Fuchs' heterochromic cyclitis (FHC) and other types of uveitis was analysed by high performance liquid chromatography (HPLC) and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Using HPLC, the number of peaks and their respective elution times were similar for the FHC, uveitis and control groups. SDS-PAGE and silver staining showed immunoglobulin G migrating as two to four distinct bands in all non-reduced samples. In the reduced state, 1-5 unidentified low MW bands (10-19 kD) were seen. Two bands at 19 and 10 kD appeared to be specific for the uveitis group, and a band at 11 kD was present in 76% of uveitis and 19% of FHC patients. None of these three bands were detected in any of the control group. As yet the identity of these low MW proteins and their possible significance in the pathogenesis of intraocular inflammation are unknown.


Subject(s)
Aqueous Humor/chemistry , Eye Proteins/analysis , Iridocyclitis/metabolism , Uveitis/metabolism , Adult , Aged , Aged, 80 and over , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel , Female , Humans , Iridocyclitis/etiology , Male , Middle Aged , Sodium Dodecyl Sulfate , Uveitis/etiology
13.
Invest Ophthalmol Vis Sci ; 31(10): 2129-35, 1990 Oct.
Article in English | MEDLINE | ID: mdl-2211009

ABSTRACT

Immunoglobulin G (IgG) in aqueous humor from patients with various uveitis syndromes was analyzed using a number of immunologic techniques. Sixty-five percent of patients with Fuchs' heterochromic cyclitis (FHC), 70% of patients with other forms of uveitis, and 44% of controls showed local synthesis of IgG, as demonstrated by an elevated IgG:albumin relative concentration ratio. Using an enzyme-linked immunosorbent assay to measure the concentration of IgG subclasses 1-4, a relative excess of IgG1 was found in the aqueous compared with the serum in FHC. Isoelectric focusing and immunoblotting studies revealed oligoclonal IgG bands in the aqueous of 13 of 23 (57%) patients with FHC, most being of the IgG1 subclass. Oligoclonal bands were not found in 18 patients with other types of uveitis or 13 patients undergoing surgery for senile cataract. These findings indicate intraocular production of IgG of restricted specificity in FHC, providing further evidence for local immune dysfunction in this condition. As yet the antigenic stimulus for this oligoclonal B-cell response has not been identified.


Subject(s)
Aqueous Humor/metabolism , Immunoglobulin G/metabolism , Uveitis/metabolism , Adult , Aged , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoblotting , Iridocyclitis/metabolism , Isoelectric Focusing , Male , Middle Aged , Reference Values , Serum Albumin/metabolism
14.
Invest Ophthalmol Vis Sci ; 31(5): 917-20, 1990 May.
Article in English | MEDLINE | ID: mdl-2335453

ABSTRACT

The level of Interleukin-6 (IL-6) in the aqueous humor of 24 patients with 2 types of uveitis was measured with a specific bioassay using the murine hybridoma cell line B9. Sixteen patients had Fuchs' heterochromic cyclitis (FHC) and 8 had toxoplasma uveitis (TU). Sixty-three percent of each of the FHC and TU groups had raised levels of IL-6 in their aqueous (mean: 543 and 19,228 units/ml respectively). Thirteen control aqueous samples, obtained at surgery for senile cataract, showed IL-6 levels of less than 10 units/ml. Serum obtained at the same time as each aqueous humor sample also showed IL-6 levels of less than 10 units/ml, indicating that the raised levels of IL-6 found in the aqueous of uveitis patients did not result from serum leakage, but from local production. This is the first report on intraocular IL-6 levels, and indicates that IL-6 may play a role as an inflammatory mediator in uveitis.


Subject(s)
Aqueous Humor/metabolism , Interleukin-6/metabolism , Iridocyclitis/metabolism , Uveitis/metabolism , Adolescent , Adult , Aged , Biological Assay , Female , Humans , Male , Middle Aged , Toxoplasmosis, Ocular/complications , Uveitis/etiology , Uveitis, Posterior/metabolism
15.
Curr Eye Res ; 9 Suppl: 53-7, 1990.
Article in English | MEDLINE | ID: mdl-2384014

ABSTRACT

Aqueous humor from 23 patients with Fuchs' heterochromic cyclitis (FHC) was analysed by a number of immunological methods. Intraocular IgG synthesis was found in 65% of patients and oligoclonal IgG bands, mainly of the IgG1 subclass, identified in 57%. There was a relative increase in IgG1 (P less than 0.01) as compared to patients with senile cataract. Local production of the cytokine Interleukin-6 was demonstrated in 63% of patients (P less than 0.01). Analysis of aqueous by HPLC and SDS-PAGE failed to reveal any abnormalities specific for FHC. These findings add further evidence to the theory of immune dysregulation in this condition.


Subject(s)
Aqueous Humor/metabolism , Iridocyclitis/metabolism , Adult , Aged , Aged, 80 and over , Albumins/metabolism , Aqueous Humor/immunology , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel , Female , Humans , Immunoblotting , Immunoglobulin G/metabolism , Immunoglobulin Isotypes , Interleukin-6/metabolism , Iridocyclitis/immunology , Male , Middle Aged , Serum Albumin/metabolism
16.
Vestn Oftalmol ; 105(5): 22-3, 1989.
Article in Russian | MEDLINE | ID: mdl-2595893

ABSTRACT

Measurements of unsaturated fatty acid diene conjugates in the anterior chamber humor are recommended to predict postoperative inflammatory complications. If their levels are increased, antioxidants should be included in the therapeutic scheme along with antibiotics and corticosteroids.


Subject(s)
Iridocyclitis/prevention & control , Postoperative Complications/prevention & control , Aqueous Humor/analysis , Cataract Extraction , Fatty Acids, Unsaturated/analysis , Humans , Iridocyclitis/metabolism , Postoperative Care/methods , Postoperative Complications/metabolism , Prognosis
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