Age-Related Maculopathy Susceptibility 2 and Complement Factor H Polymorphism and Intraocular Complement Activation in Neovascular Age-Related Macular Degeneration.

Purpose: To investigate the association of risk alleles in complement factor H (CFH) and age-related maculopathy susceptibility 2 (ARMS2) with complement activation products in the aqueous humor in eyes with neovascular age-related macular degeneration (nAMD) including polypoidal choroidal vasculopathy (PCV), retinal angiomatous proliferation (RAP), and pachychoroid neovasculopathy (PNV). Design: Prospective, comparative, observational study. Participants: Treatment-naïve patients with nAMD and cataract patients as controls. Methods: The study included 236 eyes of 236 patients with nAMD and 49 control eyes of 49 patients. Aqueous humor samples were collected from 67 eyes with drusen-associated nAMD, 72 eyes with PCV, 26 eyes with RAP, and 71 eyes with PNV before intravitreal anti-VEGF injection and cataract surgery in the 49 control eyes. Clinical samples were measured for complement component 3a (C3a), C4a, and C5a using a bead-based immunoassay. Genotyping of the ARMS2 A69S (rs10490924), CFH I62V (rs800292), and CFH Y402H (rs1061170) was performed using TaqMan genotyping. Main Outcome Measures: The levels of complement activation products (C3a, C4a, and C5a) in the aqueous humor in each genotype of ARMS2 and CFH. Results: The C3a level in the aqueous humor was significantly elevated (P = 0.006) in patients with nAMD and the ARMS2 A69S risk allele, whereas the levels of the complement activation products were not associated with CFH I62V and Y402H genotypes. Among the control eyes, no significant differences were seen in any complement activation products for all genetic polymorphisms. The levels of the complement activation products in the aqueous humor of eyes with the nAMD subtypes for each genetic polymorphism did not show significant differences. Conclusions: The C3a concentration in the aqueous humor was significantly higher in Japanese nAMD patients with the ARMS2 A69S risk allele, whereas it was not elevated in the patients with CFH I62V. Age-related maculopathy susceptibility 2 A69S polymorphism is strongly associated with local complement activation in nAMD patients.

Changes in complement activation products after anti-VEGF injection for choroidal neovascularization in age-related macular degeneration and pachychoroid disease.

We evaluated changes in the complement system resulting from anti-vascular endothelial growth factor (VEGF) in eyes with age-related choroidal neovascularization (CNV) including neovascular age-related macular degeneration, pachychoroid neovasculopathy, and polypoidal choroidal neovasculopathy. We measured the concentrations of the complement activation products (C3a, C4a), VEGF, and monocyte chemotactic protein-1 in the aqueous humor during intravitreal anti-VEGF injections for CNV. The VEGF level decreased significantly (P < 0.001), while the C3a and C4a levels increased significantly (P < 0.001 for both comparisons) 1 month after two monthly anti-VEGF injections. The VEGF level was correlated with the C3a (R = 0.328, P = 0.007) and C4a (R = - 0.237, P = 0.055) levels at baseline, but the correlation between the VEGF and C3a levels (R = - 0.148, P = 0.242) changed significantly (P = 0.028 by analysis of covariance) after anti-VEGF treatment. The C3a increase after anti-VEGF therapy did not change the visual outcomes in eyes with CNV for 1 year. Dysregulation of the complement system can be induced after anti-VEGF therapy.

Complement Activation Products and Cytokines in Pachychoroid Neovasculopathy and Neovascular Age-Related Macular Degeneration.

Purpose: To investigate the characteristics of complement activation products and angiogenic cytokines in the aqueous humor in eyes with pachychoroid neovasculopathy (PNV) and neovascular age-related macular degeneration (nAMD). Methods: This was a prospective, comparative, observational study. All patients with choroidal neovascularization were classified as PNV without polyps, PNV with polyps (polypoidal choroidal vasculopathy [PCV]), or drusen-associated nAMD according to the presence or absence of pachychoroid features and soft drusen. This study included a total of 105 eyes. Aqueous humor samples were collected from 25 eyes with PNV without polyps, 23 eyes with PCV, and 24 eyes with drusen-associated nAMD before intravitreal anti-vascular endothelial growth factor (VEGF) injection and cataract surgery in 33 control eyes. Clinical samples were measured for complement component 3a (C3a), C4a, C5a, VEGF, and macrophage chemoattractant protein 1 (MCP-1) using a bead-based immunoassay. Results: C3a and MCP-1 levels were significantly higher in PCV (P = 0.032 and P = 0.039, respectively) and drusen-associated nAMD (P = 0.01 for both comparisons) than in controls, and no difference was seen in C3a and MCP-1 levels between PNV and controls (P = 0.747 and P = 0.294, respectively). VEGF levels were significantly higher in PNV (P = 0.016), PCV (P = 0.009), and drusen-associated nAMD (P = 0.043) than in controls. In PNV, the VEGF levels elevated without elevated C3a and MCP-1. Conclusions: PNV, PCV, and drusen-associated nAMD had significantly distinct profiles of complement activation products and cytokines in the aqueous humor.

Anaphylatoxin concentration in aqueous and vitreous humor in the eyes with vitreoretinal interface abnormalities.

The complement system may be activated in the posterior segment of the eye with chorioretinal disease, which may be reflected to the concentration of anaphylatoxins in the aqueous humor. Little is known about the distribution of anaphylatoxins in the aqueous and vitreous humor. The aim of the present study was to investigate the distribution of anaphylatoxin concentration in the aqueous and vitreous humor of the eyes with idiopathic epiretinal membrane or idiopathic macular hole. This was an experimental, observational case series. This study included 43 eyes from 43 patients; 29 eyes with idiopathic epiretinal membrane, and 14 eyes with idiopathic macular hole. All 43 eyes underwent cataract surgery and vitrectomy. The aqueous and vitreous humor were collected at the surgery. The anaphylatoxin concentrations were measured by using a cytometric beads array, and the respective C3a, C4a, and C5a concentrations were 2.003 ± 0.679 (mean ± standard deviation) ng/ml, 1.389 ± 0.419 ng/ml, and 0.003 ± 0.004 ng/ml in the aqueous humor, and 1.236 ± 0.642 ng/ml, 1.250 ± 0.542 ng/ml, and 0.048 ± 0.069 ng/ml in the vitreous humor. The mean C3a concentration in the aqueous humor was significantly higher than in the vitreous humor in 43 eyes of iMH and iERM (P < 0.001). The mean C4a concentration showed no significant difference between the aqueous humor and vitreous humor (P = 0.282), and the mean C5a in the aqueous humor was significantly lower than in the vitreous humor overall (P < 0.001). The C3a concentration in the aqueous humor strongly correlated with that in the vitreous humor (R = 0.510, P < 0.001). The concentrations of C4a and C5a in the aqueous humor moderately correlated with those in the vitreous humor (C4a; R = 0.356, P = 0.019, C5a; R = 0.464, P = 0.022). In conclusion, the anaphylatoxin concentrations measured by cytometric beads array in the aqueous humor may be associated with those measured in the vitreous humor.

Evidence for Activation of Lectin and Classical Pathway Complement Components in Aqueous Humor of Neovascular Age-Related Macular Degeneration.

PURPOSE: The complement system is activated via 3 different pathways; the lectin pathway (LP), classical pathway (CP), and alternative pathway. To investigate the possible roles for the LP or CP in the development of neovascular age-related macular degeneration (nAMD), we compared aqueous humor levels of complement proteins of the LP and CP between eyes with nAMD and those with cataract as controls. METHODS: Seventeen eyes from 17 patients with treatment-naïve nAMD and 9 eyes from 9 patients with cataract were studied. Aqueous humor samples were collected before intravitreal aflibercept or ranibizumab injection for the nAMD patients and before cataract surgery for the cataract patients. Aqueous humor levels of complement C4 of the LP and CP, complement C3 of all 3 complement pathways, and mannose-binding lectin-associated serine protease (MASP)-2 of the LP were measured by enzyme-linked immunosorbent assay. Aqueous humor levels of C4a and C3a, the activation products of C4 and C3, respectively, were measured by a bead-based immunoassay. The ratios of C4a to C4 and C3a to C3, representing the degree of C4 and C3 activation, respectively, were calculated in individual patients. RESULTS: The aqueous humor levels of C4, C3, and MASP-2 were significantly lower in the nAMD eyes compared to the controls (p = 0.008, p = 0.011, and p = 0.018, respectively). In contrast, the aqueous humor levels of C4a and C3a, as well as the C4a/C4 and C3a/C3 ratios, were significantly higher in the nAMD eyes compared to the controls (p = 0.039, p = 0.003, p < 0.001, and p < 0.001, respectively). CONCLUSIONS: This study provides evidence for significant intraocular activation of either or both of the LP and CP in nAMD eyes that might be involved in the development of nAMD. The significantly lower levels of MASP-2 in the aqueous humor of the nAMD eyes were likely due to MASP-2 consumption by activation of the LP.

Near-infrared and short-wave autofluorescence in ocular specimens.

PURPOSE: To determine histopathologic characteristics of near-infrared autofluorescence (NIR-AF) and short-wave autofluorescence (SW-AF) in ocular tissue. STUDY DESIGN: Retrospective study. METHODS: Unstained specimens from four enucleated eyes with uveal melanoma were prepared for evaluation by fluorescence microscopy. The filter settings for SW-AF were 450-490 nm for excitation, 500-550 nm for emission and for NIR-AF 672.5-747.5 nm and 765-855 nm respectively. RESULTS: Hyper-SW-AF was detected in the cornea, crystalline lens, anterior border layer of the iris, basement membrane of the iris posterior epithelium, retinal pigment epithelium (RPE), Bruch's membrane, and sclera. Hyper-NIR-AF was detected in pigmented tissues, i.e., iris anterior border layer, iris posterior epithelium, ciliary pigmented epithelium, RPE, pigmented cells in the choroid and pigmented cells in the melanoma tumoral masses. The iris anterior border layer had hyper-SW-AF and hyper-NIR-AF with low magnification. The cells on the iris surface were with hyper-SW-AF; under the iris surface cells with hyper-NIR-AF were detected with high magnification. Both hyper-SW-AF and hyper-NIR-AF were in RPE cells. Pigmented cells with hyper-NIR-AF in other uveal tissues did not have hyper-SW-AF. The pigmented cells in the melanoma tumoral masses had very weak NIR-AF. CONCLUSIONS: NIR-AF was seen in the ocular pigmented tissues. The only pigmented tissue with both hyper-SW-AF and hyper-NIR-AF was RPE, the combination of which might help interpret the cellular components of fundus lesions.

A Case of Eosinophilic Granulomatosis with Polyangiitis Complicated with A IgG4 Related Disease Like Symptoms.

We report a case of eosinophilic granulomatosis with polyangiitis (EGPA) complicated with a IgG4 related disease like symptoms presenting as eyelid swellings. In the present case, the serum level of IgG4 and the ratio of IgG4 to IgG were generally increased by the disease course of EGPA. Considering the course of clinical symptoms, there is a possibility that orbital manifestations were one of the clinical features during the disease course of EGPA while the histological features of right eyelid tissue and other ocular manifestations were consistent with the diagnosis of IgG4 related disease.

Foveal structure during the induction phase of anti-vascular endothelial growth factor therapy for occult choroidal neovascularization in age-related macular degeneration.

PURPOSE: To evaluate the efficacy of monthly injections of aflibercept and ranibizumab on foveal structure after three months, for the treatment of occult choroidal neovascularization (CNV) in age-related macular degeneration (AMD). METHODS: We retrospectively studied 103 eyes with treatment-naïve neovascular AMD with occult and no classic CNV. Seventy-four of 103 eyes were treated with ranibizumab (intravitreal ranibizumab injection [IVR] group); 29 eyes were treated with aflibercept (intravitreal aflibercept injection [IAI] group). The best-corrected visual acuity and the retinal and choroidal structure at the fovea were evaluated using optical coherence tomography. RESULTS: The total foveal thickness, the height of serous retinal detachments, and subfoveal choroidal thickness were compared with baseline, and the incidence of retinal pigment epithelial elevation significantly decreased in the IAI group compared with the IVR group. In contrast, the thickness of the sensory retina at the fovea significantly decreased in the IVR group when compared with the IAI group. The logarithm of the minimum angle of resolution (logMAR) best-corrected visual acuity improved more significantly in the IVR group (-0.085±0.164) than in the IAI group (-0.020±0.125) at 3 months (P=0.017). CONCLUSION: After intravitreal injection, aflibercept more rapidly reduced subretinal fluid and subfoveal choroidal thickness. In contrast, ranibizumab decreased the sensory retinal thickness compared with aflibercept. The responses of the retinal and choroidal tissue to these anti-VEGF agents may be different during the induction phase for eyes with occult CNV secondary to neovascular AMD.

Switching to intravitreal aflibercept injection for polypoidal choroidal vasculopathy refractory to ranibizumab.

PURPOSE: To clarify the efficacy of switching to intravitreal aflibercept injection to treat polypoidal choroidal vasculopathy refractory to ranibizumab. METHODS: In this retrospective study, 43 eyes of 42 patients (mean age, 76.5 years) with polypoidal choroidal vasculopathy treated with aflibercept (2 mg/0.05 mL) were reviewed. A treatment history of 3 consecutive monthly intravitreal injections of ranibizumab as an induction phase followed by a pro re nata maintenance phase over 12 months was seen for all patients. All patients who were refractory to ranibizumab (defined as having persist subretinal or intraretinal fluid by optical coherence tomography and unchanged or decreased visual acuity compared with the time of the first ranibizumab injection, despite receiving the last 3 consecutive monthly intravitreal ranibizumab injections after 12 months). RESULTS: The mean logarithm of the minimum angle of resolution best-corrected visual acuity levels (Snellen equivalent) improved significantly (P = 0.0074) from 0.38 (20/48) at baseline to 0.34 (20/43) 3 months after switching to aflibercept (Month 3) (mean best-corrected visual acuity improvement, 0.47 line). The central retinal thickness decreased significantly (P < 0.0001) from 245 µm at baseline to 131 µm at Month 3. Of 30 eyes with polypoidal lesions at baseline, the polypoidal lesions regressed completely in 15 eyes (50%) at Month 3. CONCLUSION: Administering intravitreal aflibercept injection for patients with polypoidal choroidal vasculopathy refractory to ranibizumab maintained or improved visual acuity and reduced or eliminated exudative lesions and occluding polypoidal lesions without adverse events with short-term follow-up.

Morphologic changes of the fovea and visual acuity associated with retinal detachment secondary to circumscribed choroidal hemangioma.

PURPOSE: To clarify and review the early and late morphologic changes of the macula associating with visual loss in patients with subfoveal fluid secondary to extrafoveal circumscribed choroidal hemangiomas. METHODS: Previously six non-treated eyes of six patients with subfoveal retinal detachment secondary to extrafoveal circumscribed choroidal hemangioma were included. Visual acuity (VA), duration of visual symptoms, color fundus photography, optical coherence tomography (OCT), fundus autofluorescence, and fluorescein angiography (FA) were evaluated. RESULTS: The mean patient age was 58 years (range, 25-78). The VA and duration of symptoms in each patient was 1.2 (3 days), 0.6 (1 week), 0.4 (3 months), 0.5 (6 months), 0.02 (12 months), and 0.01 (8 years), respectively. Three patients with symptoms for less than 3 months did not have retinal pigment epithelial (RPE) alterations, retinal edema, or thinning of the retinal structure in the fovea. A patient with symptoms for 3 months had subfoveal deposits underneath the detached neurosensory retina with foveal hyperautofluorescence. Two patients with symptoms exceeding 12 months had highly affected RPE and cystoid macular degeneration. CONCLUSIONS: The VA was affected in patients with longer visual symptoms, and there are some changes in the retina and RPE in the fovea by FA and OCT. Persistent subretinal fluid secondary to choroidal hemangiomas may result in pathologic changes in the neurosensory retina.

Retinal pigment epithelium tear after intravitreal aflibercept injection.

To report a case complicated with a retinal pigment epithelium (RPE) tear after intravitreal aflibercept injection. A 78-year-old man had deteriorated visual acuity in his left eye. Fluorescein angiography showed occult choroidal neovascularization. Optical coherence tomography showed a serous retinal detachment and fibrovascular pigment epithelial detachment. He was diagnosed as typical age-related macular degeneration associated with pigment epithelial detachment and treatment consisting of three consecutive monthly intravitreal injections of aflibercept was planned. A month after the initial injection, his visual acuity had not improved. The red-free photograph showed an area of RPE defect inferior to the fovea. The fundus autofluorescence, fluorescein angiography, and optical coherence tomography clearly demonstrated the presence of an RPE tear. A second injection of aflibercept was performed due to a remaining serous retinal detachment. Although this is a single case and RPE tears may occur as a spontaneous complication of age-related macular degeneration patients, the risk of a tear should be discussed when considering aflibercept treatment for typical age-related macular degeneration patients with pigment epithelial detachment as there might be a risk for developing an RPE tear.

Autofluorescence of the cells in human subretinal fluid.

PURPOSE: The origin of autofluorescence in the subretinal space and the autofluorescence properties of the cells were investigated in surgically collected subretinal fluid. METHODS: Subretinal fluid was surgically collected from four eyes of patients with rhegmatogenous retinal detachment (three eyes) and Coats' disease (one eye). After cytocentrifuge preparation of the cells in the fluid and immunofluorescence staining, a cytologic examination was conducted by using confocal scanning laser microscopy. The autofluorescence of the cells was elucidated by measuring the fluorescence spectra with spectroscopy, to obtain different excitation laser light emission fingerprints. RESULTS: The cells from the subretinal fluid were classified into three types: CD68-negative cells containing numerous pigmented granules, CD68-positive cells containing few pigments, and CD68-negative cells with no pigmented granules. Autofluorescence was observed in the inclusions of the cells classified into the former two types. When the cells were excited by a 458- or 488-nm laser light, emission spectra in autofluorescence showed little difference between CD68-positive and -negative cells. Peak analysis confirmed that the two types of cells showed the same emission peaks within this range of excitation light. CONCLUSIONS: Autofluorescent inclusions appeared in the CD68-positive and -negative cells in the subretinal fluid. The macrophages in the subretinal fluid possess autofluorescence that is spectroscopically similar to lipofuscin. Autofluorescence of macrophages can be attributed to degenerated outer segments and debris from apoptotic photoreceptors. Clinicians should consider migration of macrophages, in addition to retinal pigment epithelium, as the possible source when abnormal fundus autofluorescence is observed using an ordinary set of fluorescence filters.