Profibrotic phenotype of conjunctival fibroblasts from mucous membrane pemphigoid.

Ocular mucous membrane pemphigoid is an immunobullous disease in which excessive conjunctival fibrosis causes blindness, and the pathogenesis of scarring is incompletely understood. To establish whether profibrotic fibroblasts with an altered phenotype exist in ocular mucous membrane pemphigoid, we compared the functional characteristics of pemphigoid conjunctival fibroblasts to normal conjunctival fibroblasts with respect to cell division; migration; collagen contraction; matrix metalloproteinase, secretion of collagen and chemokines; and myofibroblast differentiation. We found that pemphigoid fibroblasts showed increased cell division (P = 0.01), increased migration in serum-free medium (72 ± 18 migrated cells versus 33 ± 11, P = 0.04), increased collagen contraction in the presence of 10 ng/ml tumor necrosis factor-α, increased collagen type I secretion (P = 0.03), increased secretion of matrix metalloproteinase-3 (P = 0.03), and increased secretion of eotaxin in response to interleukin-13 (P = 0.04). Differences between pemphigoid and normal conjunctival fibroblasts with respect to collagen contraction and MMP secretion in the presence of interleukin-13 were also observed. Together, these findings indicate that pemphigoid conjunctival fibroblasts have a profibrotic phenotype that is maintained in vitro. No differences between pemphigoid fibroblasts obtained from acutely inflamed versus clinically uninflamed conjunctiva were observed. Developing effective antifibrotic therapies will require understanding of the mechanisms that both induce and maintain the profibrotic phenotype.

Conjunctival interleukin-13 expression in mucous membrane pemphigoid and functional effects of interleukin-13 on conjunctival fibroblasts in vitro.

Interleukin-13 (IL-13) is the dominant effector cytokine of fibrosis in pulmonary and liver disease. Excessive conjunctival fibrosis in the immunobullous disease ocular mucous membrane pemphigoid (MMP) causes blindness; the pathogenesis of scarring in this disease is incompletely understood. To determine whether IL-13 is involved in conjunctival fibrosis in MMP, we studied the expression of IL-13 in ocular MMP patients before and after systemic immunosuppression and examined the effects of IL-13 on normal human conjunctival fibroblasts. We found high stromal cell expression of IL-13 in active ocular MMP by immunohistochemistry; 80% of these cells were CD3-positive T cells. Following immunosuppression, in clinically uninflamed, treated, ocular MMP patients, the number of IL-13 positive cells was significantly reduced, but this was still fourfold greater than in normal conjunctiva. IL-13 stimulated collagen lattice contraction and migration, and decreased production of mmp-3 and mmp-10 by human conjunctival fibroblasts. The addition of T cell culture supernatant to IL-13 synergistically augmented fibroblast migration. IL-13 also up-regulated surface expression of HLA-DR, CD80, CD40, and CD154 by conjunctival fibroblasts, suggesting a potential mechanism for fibroblast-T cell cross talk, via which fibroblasts may actively engage in perpetuating chronic inflammation and continued fibrosis. Together, these findings suggest that IL-13 is involved in conjunctival fibrosis in MMP, and that IL-13 has both profibrotic and pro-inflammatory effects on human conjunctival fibroblasts.

Visual and autorefraction outcomes following toric intraocular lens insertion without calculation of posterior corneal astigmatism in the UK National Health Service.

OBJECTIVE: To ascertain visual and refractive outcomes following toric intraocular lens (IOL) implantation in the UK National Health Service (NHS) without posterior corneal astigmatism calculation, with multiple surgeons of different grades, pooled input and output pathways and autorefraction as the refractive outcome measure. METHODS: Preoperative and 1-month post-operative data were analysed retrospectively in 114 eyes (95 patients) receiving a toric IOL between 2014 and 2016 at Imperial College NHS Trust. Preoperative keratometric astigmatism was ≥2 dioptres (D). RESULTS: Mean preoperative best-corrected visual acuity (BCVA) was 0.50 logMAR (±0.46), improving to a mean uncorrected VA (UCVA) of 0.35 logMAR (±0.36) postoperatively (p < 0.001) with 65% of eyes attaining a UCVA ≤ 0.30 logMAR. Excluding 33 eyes with pre-existing visual comorbidities and one targeting monovision, mean post-operative UCVA was 0.24 logMAR (±0.29) (p < 0.001), and 85% had UCVA ≤ 0.30 logMAR, 62% UCVA ≤ 0.20 logMAR. Mean refractive astigmatism improved from 3.04 D (±1.46) to 1.36 D (±1.13) (p < 0.001). In total, 52% of eyes had post-operative refractive astigmatism ≤1.00 D. The Alpins correction index was 1.05 (±0.22), indicating a tendency to overcorrect. Toric IOL misalignment was noted in two eyes, and two cases of posterior capsule rupture were converted to a non-toric IOL. CONCLUSIONS: Visual outcomes of toric IOL implantation in our pooled pathway are comparable to single-surgeon case series where posterior corneal astigmatism has not been accounted for. However, with 1-month post-operative autorefraction, only 52% of eyes had ≤1 D refractive astigmatism, which is lower than previously published series, but may be standard for 1-month autorefraction outcomes.

Immunosuppressive therapy for ocular mucous membrane pemphigoid strategies and outcomes.

PURPOSE: To evaluate the effectiveness and toxicity of a stepladder immunosuppression strategy, including the use of mycophenolate mofetil and combination therapy, in the treatment of ocular mucous membrane pemphigoid. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Two hundred twenty-three eyes of 115 patients. METHODS: Patients with a diagnosis of ocular mucous membrane pemphigoid commencing immunosuppression between January 1994 and July 2005 were identified. A treatment episode was defined by the use of a particular therapy or combination of therapies. MAIN OUTCOME MEASURES: For each treatment episode, success of immunosuppressive therapy in controlling ocular inflammation was graded as a success (S), qualified success (QS), or failure (F). Initial and final visual acuities (VAs), stage of cicatrization (Foster, Mondino), grade of conjunctival inflammation, and side effects were recorded. RESULTS: In 70% (80/115) of patients, inflammation was controlled by the end of the study. At least 6 months remission off treatment occurred in 16 patients (14%). Of the 388 treatment episodes, 50% were classified as S; 27%, QS; and 23%, F. The most successful therapies were based on cyclophosphamide (S, 69%; QS, 21%; F, 10%), followed by mycophenolate (S, 59%; QS, 22%; F, 19%), azathioprine (S, 47%; QS, 24%; F, 29%), dapsone (S, 47%; QS, 30%; F, 23%), and sulfapyridine (S, 38%; QS, 27%; F, 35%). Combination sulfa-steroid-myelosuppressive agent therapy increased the response from 73% with single-agent therapy to 87%. Side effects were the reason for 29% of changes in therapy. These were most prominent with azathioprine (40%) and least with mycophenolate (15%). Initial best-corrected VA (BCVA) was 6/60 or less in 17% (37/223) of eyes, pemphigoid being the cause in 13% (29/223). Final BCVA was 6/60 or less in 34% (76/223) of eyes, pemphigoid being the cause in 26% (57/223). By the end of the study, Mondino stage cicatrization had progressed in 41% (92/223) of eyes and 53% (61/115) of patients. CONCLUSIONS: Mycophenolate mofetil seems to be an effective and well-tolerated immunosuppressant for moderately active ocular mucous membrane pemphigoid. Combination sulfa-steroid-myelosuppressive agent therapy in a stepladder regimen is a useful strategy to improve disease control. Cicatrization and VA may still progress and worsen despite adequate control of inflammation.

Ocular mucous membrane pemphigoid: diagnosis and management strategies.

Ocular mucous membrane pemphigoid presents some of the most challenging dilemmas in anterior segment management. Diagnosis is made difficult by the insensitivity of immunopathological investigations and the differential diagnosis of other scarring conjunctival disorders. The management of the associated ocular surface disease involves control of blepharitis, dry eye, filamentary keratitis, keratinization, lid malposition, and persistent epithelial defect, as well as the identification and avoidance of toxicity. Inflammation associated with the underlying disorder demands the use of systemic immunosuppressive therapy in many patients. New biological immunotherapies have been used when conventional immunosuppressive therapies fail. Ophthalmic plastic surgery is essential for the management of lid malposition and corneal exposure. Improving vision may require the use of specialized contact lenses, cataract surgery, and corneal and ocular surface reconstructive surgery. All surgery must be integrated with ocular surface treatment and immunosuppressive treatment to avoid disease exacerbations.

The natural history of Stevens Johnson syndrome: patterns of chronic ocular disease and the role of systemic immunosuppressive therapy.

OBJECTIVE: To characterize patterns of chronic ocular disease in patients with Stevens-Johnson syndrome (SJS) and its variant toxic epidermal necrolysis (TEN), and to describe their response to treatment. METHODS: Retrospective case series. A review of hospital records of 30 patients (60 eyes) with ocular manifestations of SJS or TEN was carried out. The principal outcome measure was to identify and classify the patterns of chronic ocular disease in SJS and TEN. The secondary outcome measure was the response to treatment. RESULTS: Patterns of chronic ocular disease observed after the acute episode included: mild/moderate SJS, severe SJS, ocular surface failure (SJS-OSF), recurrent episodic inflammation (SJS-RI), scleritis (SJS-S) and progressive conjunctival cicatrisation resembling mucous membrane pemphigoid (SJS-MMP). The median follow-up was 5 years (range 0-29). 19 patients (29 eyes (48%)) developed SJS-OSF, SJS-RI, SJS-S or SJS-MMP during follow-up. SJS-OSF was present in 12 patients (18 eyes (30%)). In 5 patients (eight eyes) this developed 1 year after the acute illness, without any further inflammatory episodes; it was associated with SJS-RI in 1 patient (2 eyes), with SJS-RI and SJS-S in 1 patient (1 eye), with SJS-S in 1 patient (1 eye) and with SJS-MMP in 4 patients (6 eyes). Episodes of SJS-RI occurred in 4 patients (7 eyes (12%)). The median time from acute disease to the first episode of SJS-RI was 8.5 years (range 5-63). SJS-S developed in 2 patients (4 eyes (7%)), of which 2 eyes subsequently developed SJS-OSF. SJS-MMP developed in 5 patients (10 eyes (16.6%)). The median duration from the acute stage to the diagnosis of SJS-MMP was 2 years (range 1-14). Immunosuppressive therapy successfully controlled inflammation in 10/10 patients with SJS-MMP, SJS-RI or SJS-S. CONCLUSION: Ocular disease in SJS/TEN is not limited solely to the sequelae of the acute phase illness. Patients and physicians need to know that ocular disease progression, due to surface failure and/or acute inflammatory conditions, may occur at variable periods following the acute disease episode. Recognition of this, and prompt access to specialist services, may optimise management of these uncommon patterns of disease in SJS.

Amniotic membrane transplantation for ocular disease: a review of the first 233 cases from the UK user group.

BACKGROUND: Amniotic membrane transplantation (AMT), as a new tool in the armamentarium of therapies available for ocular surface problems, became widely available in the UK in 1998. This study evaluates the indications for treatment, the surgical procedures used, and the results of a subset of the first AMT cases carried out by the group using this nationally available supply. This user group model provides data which is different from that obtained from uncontrolled case series, or clinical trials, and may be more representative of the outcomes that can be expected when a procedure becomes widely available. METHODS: The first 233 AMTs, performed by the UK user group, were evaluated by audit and outcomes were assessed at 3 months. RESULTS: Of the 233 transplants, there were 126 (54.1%) valid outcome returns: the outcome for persistent epithelial defects was a healed and stable surface in 11/35 (31.4%, 95% CI 16.9 to 49.3); for chemical/thermal injuries, a healed uninflamed eye with clear cornea in 5/18 (27.8%, 95% CI 9.7 to 53.4); for bullous keratopathy a pain-free, stable surface without bullae in 4/18 (22.2%, 95% CI 6.4 to 47.6); for ocular surface reconstruction, an epithelialised uninflamed conjunctiva without scarring in 12/23 (52.2%, 95% CI 30.6 to 73.2); and for limbal stem cell deficiency, a corneal phenotype in 4/7 (57.1%). The operative technique least associated with failure was use of a bandage contact lens at the end of the procedure (OR 0.19, 95% CI 0.06 to 0.59, p = 0.004). Previous treatment with topical steroids was significantly associated with failure (OR 5.70, 95% CI 1.77 to 18.43, p = 0.004). CONCLUSION: Although the outcome criteria used in this study were stringent, and the follow-up duration was short, the results of AMT by this user group were generally less favourable than those of previously reported case series. Controlled clinical trials would improve the quality of evidence for use of amniotic membrane in ocular disease.

Deep anterior lamellar keratoplasty using the manual dissection technique of Melles: a histopathologic correlation.

PURPOSE: To report the histopathologic findings in the host tissue of 2 human keratoconic corneas undergoing maximum-depth anterior lamellar keratoplasty (MDALK) using the manual dissection technique described by Melles. METHODS: Corneal buttons were examined from 2 patients with keratoconus who underwent MDALK using the manual dissection technique of Melles and converted to penetrating keratoplasty after rupture of the lamellar bed. Manual dissection was performed in 1 patient, and combined manual and viscoelastic dissection of Descemet membrane (DM) was performed in the other. RESULTS: Light microscopy of the corneal buttons showed a deep pre-Descemet dissection plane with minimal residual stroma. DM appeared to be thinned in both eyes and measured 3 to 8 microm in thickness. CONCLUSION: By using the manual dissection technique of Melles, LK can be performed exposing the smooth DM of the recipient bed. We confirmed exposure of DM in patient corneas, using this technique by light microscopy. There may be an increased risk of rupture of DM during surgery when this membrane is thinned, particularly in patients with keratoconus. This surgical technique allows conversion to penetrating keratoplasty after rupture of DM.

Validation of a fornix depth measurer: a putative tool for the assessment of progressive cicatrising conjunctivitis.

BACKGROUND/AIMS: Documentation of conjunctival forniceal foreshortening in cases of progressive cicatrising conjunctivitis (PCC) is important in ascertaining disease stage and progression. Lower fornix shortening is often documented subjectively or semi-objectively, whereas upper forniceal obliteration is seldom quantified. Although tools such as fornix depth measurers (FDMs) have been described, their designs limit upper fornix measurement. The purpose of this study was to custom-design a FDM to evaluate the upper fornix and to assess variability in gauging fornix depth. METHODS: A polymethylmethacrylate FDM was constructed using industry-standard jewellery computer software and machinery. Two observers undertook a prospective independent evaluation of central lower fornix depth in a heterogeneous cohort of patients with clinically normal and abnormal conjunctival fornices both subjectively and by using the FDM (in mm). Upper central fornix depth was also measured. Agreement was assessed using Bland-Altman plots. RESULTS: Fifty-one eyes were evaluated. There was 100% intraobserver agreement to within 1 mm for each observer for lower fornix measurement. The mean difference in fornix depth loss using the FDM between observer 1 and 2 was 1.19%, with 95% confidence of agreement (±2SD) of -15% to +20%. In total, 86% (44/51) of measurements taken by the two observers agreed to within 10% of total lower fornix depth (ie, ±1 mm) versus only 63% (32/51) of the subjective measurements. Mean upper fornix difference was 0.57 mm, with 95% confidence of agreement of between -2 and +3 mm. CONCLUSIONS: This custom-designed FDM is well tolerated by patients and shows low intraobserver and interobserver variability. This enables repeatable and reproducible measurement of upper and lower fornix depths, facilitating improved rates of detection and better monitoring of progression of conjunctival scarring.

Acanthamoeba keratitis: diagnosis and treatment update 2009.

PURPOSE: To describe the current management of Acanthamoeba keratitis (AK). DESIGN: A perspective based on the literature and author experience. RESULTS: Early diagnosis and appropriate therapy are key to a good prognosis. A provisional diagnosis of AK can be made using the clinical features and confocal microscopy, although a definitive diagnosis requires culture, histology, or identification of Acanthamoeba deoxyribonucleic acid by polymerase chain reaction. Routine use of tissue diagnosis is recommended, particularly for patients unresponsive to treatment for AK. Topical biguanides are the only effective therapy for the resistant encysted form of the organism in vitro, if not always in vivo. None of the other drugs that have been used meet the requirements of consistent cysticidal activity and may have no therapeutic role. The use of topical steroids is controversial, but probably beneficial, for the management of severe corneal inflammatory complications that have not responded to topical biguanides alone. The scleritis associated with AK is rarely associated with extracorneal invasion and usually responds to systemic anti-inflammatory treatment combined with topical biguanides. Therapeutic keratoplasty retains a role for therapy of some severe complications of AK but not for initial treatment. With modern management, 90% of patients can expect to retain visual acuity of 6/12 or better and fewer than 2% become blind, although treatment may take 6 months or more. CONCLUSIONS: Better understanding of the pathogenesis of the extracorneal complications, the availability of polymerase chain reaction for tissue diagnosis, and effective licensed topical anti-amoebics would substantially benefit patients with AK.

Tumor necrosis factor-alpha in ocular mucous membrane pemphigoid and its effect on conjunctival fibroblasts.

PURPOSE: First, to determine whether tumor necrosis factor-(TNF)-alpha is expressed in the conjunctiva of ocular mucous membrane pemphigoid (MMP) and the consequences of systemic immunosuppressive treatment on this expression. Second, to investigate the in vitro effects of TNFalpha on human conjunctival fibroblasts. METHODS: The expression of TNFalpha in conjunctival tissues of patients with actively inflamed ocular MMP (n = 10), patients with clinically noninflamed ocular MMP after systemic immunosuppressive treatment (n = 10), and normal subjects (n = 10) was studied by immunohistochemistry. The effect of TNFalpha on functional assays of human conjunctival fibroblast activity were investigated, including migration, collagen lattice contraction, matrix metalloproteinase (mmp), and tissue inhibitor of matrix metalloproteinase (timp) secretion, proliferation, and surface expression of HLA-DR, ICAM, CD80, CD86, CD40, CD40-ligand. RESULTS: In active ocular MMP, TNFalpha is expressed by a large number of stromal infiltrating cells (234 cells/mm(2)), and although the level of stromal TNFalpha expression is significantly reduced after immunosuppressive treatment (90 cells/mm(2)), these levels are still significantly elevated compared with normal conjunctiva (10 cells/mm(2), P < 0.05). TNFalpha stimulates increased migration by conjunctival fibroblasts (P < 0.001), increased production of mmp-9 (P = 0.01), decreased production of timp-2 (P = 0.01) and timp-4 (P = 0.04), and upregulated expression of CD40 and ICAM (P = 0.04). No significant effects of TNFalpha on fibroblast proliferation or collagen lattice contraction were detected. CONCLUSIONS: Increased conjunctival expression of TNFalpha in ocular MMP suggests that systemic TNFalpha antagonists are likely to be effective in controlling severe disease unresponsive to conventional systemic immunosuppression. Residual TNFalpha expression persists in clinically noninflamed disease. TNFalpha appears to have profibrotic and proinflammatory effects on human conjunctival fibroblasts.