Eric A. Steele

IgG4 Immunostaining and Its Implications in Orbital Inflammatory Disease

Objective IgG4-related disease is an emerging clinical entity which frequently involves tissue within the orbit. In order to appreciate the implications of IgG4 immunostaining, we analyzed gene expression and the prevalence of IgG4- immunostaining among subjects with orbital inflammatory diseases. Methods We organized an international consortium to collect orbital biopsies from 108 subjects including 22 with no known orbital disease, 42 with nonspecific orbital inflammatory disease (NSOI), 26 with thyroid eye disease (TED), 12 with sarcoidosis, and 6 with granulomatosis with polyangiitis (GPA). Lacrimal gland and orbital adipose tissue biopsies were immunostained for IgG4 or IgG secreting plasma cells. RNA transcripts were quantified by Affymetrix arrays. Results None of the healthy controls or subjects with TED had substantial IgG4 staining. Among the 63 others, the prevalence of significant IgG4-immunostaining ranged from 11 to 39% depending on the definition for significant. IgG4 staining was detectable in the majority of tissues from subjects with GPA and less commonly in tissue from subjects with sarcoidosis or NSOI. The detection of IgG4+ cells correlated with inflammation in the lacrimal gland based on histology. IgG4 staining tissue expressed an increase in transcripts associated with inflammation, especially B cell-related genes. Functional annotation analysis confirmed this. Conclusion IgG4+ plasma cells are common in orbital tissue from patients with sarcoidosis, GPA, or NSOI. Even using the low threshold of 10 IgG4+ cells/high powered field, IgG4 staining correlates with increased inflammation in the lacrimal gland based on histology and gene expression.

Müller muscle-conjunctiva resection to correct ptosis in high-risk patients.

Purpose: Müller muscle-conjunctiva resection could be seen as a relative contraindication in patients with a prior history of a glaucoma filtering procedure, corneal disease, or corneal surgery. The concern centers around the theoretical risk of bleb-related complications or corneal damage from the palpebral conjunctival sutures. Our study aimed to determine whether any bleb- or cornea-related complications arose in patients who underwent Müller muscle-conjunctiva resection for ptosis correction. Methods: A retrospective chart review was performed on 2 practices of oculofacial plastic surgeons from 2000 to 2006, including patients who had ptosis correction by Müller muscle-conjunctiva resection. Patients with a prior history of a glaucoma filtering procedure, corneal disease, or corneal surgery were identified. Each case was reviewed to determine whether any bleb- or cornea-related complications occurred. The postoperative improvement of ptosis measured by interpalpebral distance or margin reflex distance-1 also was noted. Results: Forty-three patients and 55 eyes with a history of a glaucoma filtering procedure (13 patients/15 eyes), corneal disease (1 patient/1 eye), or corneal surgery (29 patients/39 eyes) who underwent Müller muscle-conjunctiva resection were identified. The average follow-up time was 212.4 days. No bleb-related complications occurred. One patient with a history of Reis-Bücklers dystrophy experienced a corneal abrasion. Fifty-two of 55 patients had objective improvement of their ptosis. Conclusions: Müller muscle-conjunctiva resection can provide an effective means for ptosis repair in patients with a prior history of a glaucoma filtering procedure, corneal disease, or corneal surgery. One temporary postoperative corneal complication occurred in our series.

Comparison of injection pain of articaine and lidocaine in eyelid surgery

Purpose: To compare the pain induced by tissue infiltration of lidocaine 2% with epinephrine 1:100,000 versus articaine 4% with epinephrine 1:100,000 for eyelid surgery. Methods: Thirty patients undergoing bilateral eyelid surgery were enrolled in a prospective, randomized, double-masked study. Each subject received injections of lidocaine 2% with epinephrine 1:100,000 (Xylocaine) on one side and articaine 4% with epinephrine 1:100,000 (Septocaine) on the other for surgical anesthesia. The patients rated the pain of infiltration using a 100-mm visual analogue scale immediately after receiving each injection. The pain scores were compared using the paired t test. Results: Twenty-two of the 30 patients (73.3%) rated the articaine injection as less painful than the lidocaine injection. The mean pain score for lidocaine was 42.60 ± 24.74 and the pain score for articaine was 31.85 ± 20.28 (p = 0.011). Conclusions: In this study, infiltration of articaine was less painful than lidocaine for eyelid surgery, making articaine an attractive alternative for local anesthesia.

Conjunctivodacryocystorhinostomy with the frosted jones pyrex tube.

Purpose: To report the results of conjunctivodacryocystorhinostomy with primary placement of a frosted Jones Pyrex tube in the treatment of epiphora from upper lacrimal dysfunction. Methods: A retrospective chart review was performed for patients who had undergone conjunctivodacryocystorhinostomy with primary placement of a frosted Jones Pyrex tube performed by a single surgeon (R.A.D.). All patients with at least 6 months of follow-up were included in the study. Efficacy was judged by patient report of resolution of tearing and charts were reviewed for complications. Results: Five conjunctivodacryocystorhinostomy procedures were performed with primary placement of a frosted Jones Pyrex tube. Four of the surgeries were performed for a diagnosis of flaccid canaliculi and one for a congenital upper lacrimal obstruction that had been unsuccessfully treated with previous surgery at another institution. Three of the surgeries were performed with an endoscopic approach and 2 were performed with an external approach. Follow-up ranged from 29 to 34 weeks (mean, 31.2 weeks). All patients reported complete resolution of tearing and no complications were noted, including no evidence of tube migration or extrusion. Conclusions: Primary placement of frosted Jones Pyrex tubes in patients undergoing conjunctivodacryocystorhinostomy seems to retain the efficacy of a standard Jones Pyrex tube while reducing the likelihood of tube extrusion, which is the main complication of this surgery.

Orbital pseudotumor can be a localized form of granulomatosis with polyangiitis as revealed by gene expression profiling.

Biopsies and ANCA testing for limited forms of granulomatosis with polyangiitis (GPA) are frequently non-diagnostic. We characterized gene expression in GPA and other causes of orbital inflammation. We tested the hypothesis that a sub-set of patients with non-specific orbital inflammation (NSOI, also known as pseudotumor) mimics a limited form of GPA. Formalin-fixed, paraffin-embedded orbital biopsies were obtained from controls (n=20) and patients with GPA (n=6), NSOI (n=25), sarcoidosis (n=7), or thyroid eye disease (TED) (n=20) and were divided into discovery and validation sets. Transcripts in the tissues were quantified using Affymetrix U133 Plus 2.0 microarrays. Distinct gene expression profiles for controls and subjects with GPA, TED, or sarcoidosis were evident by principal coordinate analyses. Compared with healthy controls, 285 probe sets had elevated signals in subjects with GPA and 1472 were decreased (>1.5-fold difference, false discovery rate adjusted p<0.05). The immunoglobulin family of genes had the most dramatic increase in expression. Although gene expression in GPA could be readily distinguished from gene expression in TED, sarcoidosis, or controls, a comparison of gene expression in GPA versus NSOI found no statistically significant differences. Thus, forms of orbital inflammation can be distinguished based on gene expression. NSOI/pseudotumor is heterogeneous but often may be an unrecognized, localized form of GPA.

Dermis fat graft implantation after unilateral enucleation for retinoblastoma in pediatric patients.

Purpose: To evaluate the efficacy of dermis fat graft (DFG) as a primary implant technique in pediatric patients requiring unilateral enucleation due to retinoblastoma. Methods: A retrospective chart review of 14 consecutive pediatric patients who underwent dermis fat graft implantation after unilateral enucleation for retinoblastoma by 1 surgeon (E.A.S.) was performed to evaluate graft efficacy with regard to orbital volume growth and any associated morbidity. Patients who received chemotherapy or external beam radiation were excluded. Demographic information was recorded. Serial MRIs were used to measure orbital volumes to compare the surgical and contralateral orbits over time. The main outcome measure was the difference in bony orbital volume between enucleated and contralateral, uninvolved orbits. Mann-Whitney U test was used to compare orbital volume measurements between surgical and nonsurgical orbits. Correlation testing was performed to determine the effect of age, sex, and follow-up time on the orbital volume changes. Results: There was no statistical difference between the MRI volume measured for surgical and nonsurgical orbits over time. This was the case at all measured time points and for all ages and genders. All patients were under the age of 4 years at the time of surgery. The median difference in orbital volumes between surgical and nonsurgical orbits was −0.095 cm3 (range −1.26 to 1.01 cm3; quartiles −0.32 to 0.07 cm3; mean ± SD, −0.144 ± 0.0522 cm3; 95% confidence interval, −0.247 to −0.0419 cm3). The median follow-up time from surgery date to the most recent clinical examination was 38.5 months (range, 13 to 70 months; quartiles, 28.75 to 45.5 months; mean ± standard deviation [SD], 38.43 ± 17.21 months; 95% confidence interval, 29.41 to 47.45 months). Conclusions: In pediatric patients below 4 years of age with unilateral retinoblastoma treated with enucleation and primary dermis fat graft implantation, there was no statistically significant difference in bony orbital volume between the surgical and nonsurgical orbits during the follow-up period.

Parallel Gene Expression Changes in Sarcoidosis Involving the Lacrimal Gland, Orbital Tissue, or Blood

IMPORTANCE Sarcoidosis is a major cause of ocular or periocular inflammation. The pathogenesis of sarcoidosis is incompletely understood and diagnosis often requires a biopsy. OBJECTIVE To determine how gene expression in either orbital adipose tissue or the lacrimal gland affected by sarcoidosis compares with gene expression in other causes of orbital disease and how gene expression in tissue affected by sarcoidosis compares with gene expression in peripheral blood samples obtained from patients with sarcoidosis. DESIGN, SETTING, AND PARTICIPANTS In a multicenter, international, observational study, gene expression profiling of formalin-fixed biopsy specimens, using GeneChipp U133 Plus 2 microarrays (Affymetrix), was conducted between October 2012 and January 2014 on tissues biopsied from January 2000 through June 2013. Participants included 12 patients with orbital sarcoidosis (7 in adipose tissue; 5 affecting the lacrimal gland) as well as comparable tissue from 6 healthy individuals serving as controls or patients with thyroid eye disease, nonspecific orbital inflammation, or granulomatosis with polyangiitis. In addition, results were compared with gene expression in peripheral blood samples obtained from 12 historical individuals with sarcoidosis. MAIN OUTCOMES AND MEASURES Significantly differentially expressed transcripts defined as a minimum of a 1.5-fold increase or a comparable decrease and a false discovery rate of P < .05. RESULTS Signals from 2449 probe sets (transcripts from approximately 1522 genes) were significantly increased in the orbital adipose tissue from patients with sarcoidosis. Signals from 4050 probe sets (approximately 2619 genes) were significantly decreased. Signals from 3069 probe sets (approximately 2001 genes) were significantly higher and 3320 (approximately 2283 genes) were significantly lower in the lacrimal gland for patients with sarcoidosis. Ninety-two probe sets (approximately 69 genes) had significantly elevated signals and 67 probe sets (approximately 56 genes) had significantly lower signals in both orbital tissues and in peripheral blood from patients with sarcoidosis. The transcription factors, interferon-response factor 1, interferon-response factor 2, and nuclear factor κB, were strongly implicated in the expression of messenger RNA upregulated in common in the 3 tissues. CONCLUSIONS AND RELEVANCE Gene expression in sarcoidosis involving the orbit or lacrimal gland can be distinguished from gene expression patterns in control tissue and overlaps with many transcripts upregulated or downregulated in the peripheral blood of patients with sarcoidosis. These observations suggest that common pathogenic mechanisms contribute to sarcoidosis in different sites. The observations support the hypothesis that a pattern of gene expression profiles could provide diagnostic information in patients with sarcoidosis.

Fibrosis, gene expression and orbital inflammatory disease

Background/aims To clarify the pathogenesis of fibrosis in inflammatory orbital diseases, we analysed the gene expression in orbital biopsies and compared our results with those reported for idiopathic pulmonary fibrosis. Methods We collected 140 biopsies from 138 patients (58 lacrimal glands; 82 orbital fat). Diagnoses included healthy controls (n=27), non-specific orbital inflammation (NSOI) (n=61), thyroid eye disease (TED) (n=29), sarcoidosis (n=14) and granulomatosis with polyangiitis (GPA) (n=7). Fibrosis was scored on a 0–3 scale by two experts, ophthalmic pathologists. Gene expression was quantified using Affymetrix U133 plus 2.0 microarray. Results Within orbital fat, fibrosis was greatest among subjects with GPA (2.75±0.46) and significantly increased in tissue from subjects with GPA, NSOI or sarcoidosis (p<0.01), but not for TED, compared with healthy controls (1.13±0.69). For lacrimal gland, the average score among controls (1.36±0.48) did not differ statistically from any of the four disease groups. Seventy-three probe sets identified transcripts correlating with fibrosis in orbital fat (false discovery rate <0.05) after accounting for batch effects, disease type, age and sex. Transcripts with increased expression included fibronectin, lumican, thrombospondin and collagen types I and VIII, each of which has been reported upregulated in pulmonary fibrosis. Conclusions A pathologists recognition of fibrosis in orbital tissue correlates well with increased expression of transcripts that are considered essential in fibrosis. Many transcripts implicated in orbital fibrosis have been previously implicated in pulmonary fibrosis. TED differs from other causes of orbital fat inflammation because fibrosis is not a major component. Marked fibrosis is less common in the lacrimal gland compared with orbital adipose tissue.

The Role of the Immune Response in the Pathogenesis of Thyroid Eye Disease: A Reassessment.

Background Although thyroid eye disease is a common complication of Graves’ disease, the pathogenesis of the orbital disease is poorly understood. Most authorities implicate the immune response as an important causal factor. We sought to clarify pathogenesis by using gene expression microarray. Methods An international consortium of ocular pathologists and orbital surgeons contributed formalin fixed orbital biopsies. RNA was extracted from orbital tissue from 20 healthy controls, 25 patients with thyroid eye disease (TED), 25 patients with nonspecific orbital inflammation (NSOI), 7 patients with sarcoidosis and 6 patients with granulomatosis with polyangiitis (GPA). Tissue was divided into a discovery set and a validation set. Gene expression was quantified using Affymetrix U133 Plus 2.0 microarrays which include 54,000 probe sets. Results Principal component analysis showed that gene expression from tissue from patients with TED more closely resembled gene expression from healthy control tissue in comparison to gene expression characteristic of sarcoidosis, NSOI, or granulomatosis with polyangiitis. Unsupervised cluster dendrograms further indicated the similarity between TED and healthy controls. Heat maps based on gene expression for cytokines, chemokines, or their receptors showed that these inflammatory markers were associated with NSOI, sarcoidosis, or GPA much more frequently than with TED. Conclusion This is the first study to compare gene expression in TED to gene expression associated with other causes of exophthalmos. The juxtaposition shows that inflammatory markers are far less characteristic of TED relative to other orbital inflammatory diseases.

Conjunctivodacryocystorhinostomy with Jones tube: a history and update

Purpose of review Current opinions and trends in the management of upper lacrimal obstruction include design variations on the original Lester Jones tube and updated awareness and management of the problems associated with the tubes. This article includes a brief review of the fascinating history of the development of the Jones tube, which sets the perspective for the current scientific dialog. Recent findings First, many design modifications have been proposed to reduce the risk of tube migration and extrusion, with no consensus on the best tube. Second, the issue of retrograde airflow through the Jones tube with the use of continuous positive airway pressure is an increasingly common and challenging problem. Third, bacterial biofilms on the surface of the Jones tube can play a role in recalcitrant infections. Jones tubes can be cleaned or replaced in the office setting with topical anesthesia. Summary Conjunctivodacryocystorhinostomy (CDCR) with placement of a Jones tube remains the gold standard for management of upper lacrimal obstruction. This article provides an updated perspective on issues with extrusion or migration of the tube, bothersome retrograde airflow with the use of a continuous positive airway pressure device, and management of crusting and possible infectious biofilms on the tube.