Cytokine analysis of aqueous humor in patients with cytomegalovirus corneal endotheliitis.

PURPOSE: To evaluate cytokine levels of aqueous humor in patients with cytomegalovirus (CMV) corneal endotheliitis and their relationships with CMV DNA load. METHODS: 44 aqueous humor samples were obtained from 26 patients with CMV corneal endotheliitis at various stages of treatment. 33 samples obtained from cataract patients during the same period were selected as a control group. Each sample was used to measure the concentration of the CMV DNA load using real-time quantitative polymerase chain reaction, and to examine the levels of IL-6, IL-8, IL-10, MCP-1, VCAM-1, VEGF, IP-10, G-CSF, ICAM-1 and IFN-γ using a cytometric bead array. RESULTS: All 10 cytokines were found to have statistically significant differences between the CMV endotheliitis and cataract groups. The Spearman correlation test showed that the concentration of CMV DNA load was significantly associated with the levels of IL-6 (P = 0.005, r = 0.417), IL-8 (P < 0.001, r = 0.514), IL-10 (P < 0.001, r = 0.700), MCP-1 (P = 0.001, r = 0.487), VEGF (P < 0.001, r = 0.690), IP-10 (P = 0.001, r = 0.469), G-CSF (P < 0.001, r = 0.554) and ICAM-1 (P < 0.001, r = 0.635), but not significantly associated with VCAM-1 (P = 0.056) and IFN-γ (P = 0.219). CONCLUSIONS: There was a combined innate and adaptive immune response in aqueous humor in patients with CMV endotheliitis. Levels of multiple cytokines were significantly correlated with viral particle. Cytokines are potential indicators to help diagnose CMV endotheliitis, evaluate disease activity and assess treatment response.

Role Played by Receptors for Advanced Glycosylation End Products in Corneal Endothelial Cells after HSV-1 Infection.

Senescence, sterile inflammation, and infection cause dysfunction of corneal endothelial cells, leading to visual morbidity that may require corneal transplantation. With increasing age, the extracellular matrix is modified by non-enzymatic glycation forming advanced glycation end products (AGEs). The modifications are primarily sensed by the receptors for the AGEs (RAGE) and are manifested as a type I interferon response. Interestingly, in our study, human corneal endothelial cells (HCEn) cells did not respond to the typical RAGE ligands, including the AGEs, high mobility group box 1 (HMGB1), and serum amyloid-A (SAA). Instead, HCEn cells responded exclusively to the CpG DNA, which is possessed by typical corneal pathogen, herpes simplex virus-1 (HSV-1). Upon HSV-1 infection, the surface expression of RAGE was increased, and endocytosed HSV-1 was associated with RAGE and CpG DNA receptor, TLR9. RAGE DNA transfection markedly increased interferon-ß secretion by CpG DNA or HSV-1 infection. HSV-1 infection-induced interferon-ß secretion was abolished by TLR9 inhibition and partially by RAGE inhibition. Global transcriptional response analysis confirmed that RAGE and TLR9 were both significantly involved in type I interferon responses. We conclude that RAGE is a sensor of HSV-1 infection and provokes a type I interferon response.

Lassa Virus Targeting of Anterior Uvea and Endothelium of Cornea and Conjunctiva in Eye of Guinea Pig Model.

Lassa virus (LASV), a hemorrhagic fever virus endemic to West Africa, causes conjunctivitis in patients with acute disease. To examine ocular manifestations of LASV, we histologically examined eyes from infected guinea pigs. In fatal disease, LASV immunostaining was most prominent in the anterior uvea, especially in the filtration angle, ciliary body, and iris and in and around vessels in the bulbar conjunctiva and peripheral cornea, where it co-localized with an endothelial marker (platelet endothelial cell adhesion molecule). Antigen was primarily associated with infiltration of T-lymphocytes around vessels in the anterior uvea and with new vessel formation at the peripheral cornea. In animals that exhibited clinical signs but survived infection, eyes had little to no inflammation and no LASV immunostaining 6 weeks after infection. Overall, in this model, LASV antigen was restricted to the anterior uvea and was associated with mild chronic inflammation in animals with severe disease but was not detected in survivors.

Pathogenesis, Transmissibility, and Tropism of a Highly Pathogenic Avian Influenza A(H7N7) Virus Associated With Human Conjunctivitis in Italy, 2013.

H7 subtype influenza viruses represent a persistent public health threat because of their continued detection in poultry and ability to cause human infection. An outbreak of highly pathogenic avian influenza H7N7 virus in Italy during 2013 resulted in 3 cases of human conjunctivitis. We determined the pathogenicity and transmissibility of influenza A/Italy/3/2013 virus in mouse and ferret models and examined the replication kinetics of this virus in several human epithelial cell types. The moderate virulence observed in mammalian models and the capacity for transmission in a direct contact model underscore the need for continued study of H7 subtype viruses.

Corneal endothelial cells activate innate and acquired arm of anti-viral responses after cytomegalovirus infection.

Infection of the corneal endothelial cells by human cytomegalovirus (CMV) is an important cause of corneal endotheliitis. CMV endotheliitis is difficult to completely cure and relapses are frequent. This can cause blinding corneal bullous keratopathy. However, the pathogenesis of CMV endotheliitis remains undetermined. To understand the immunopathology of endotheliitis, we examined how corneal endothelial cells prime the anti-viral immunity after CMV infection based on global transcriptional responses. To accomplish this, human corneal endothelial (HCEn) cells were infected with CMV, and the global transcriptional responses were determined by microarray analyses for primary anti-viral responses using network analysis. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and protein array analyses were used to examine whether anti-viral cytokines were induced, i.e., to determine whether innate immune responses were activated. To examine whether priming of acquired immune response was activated, CMV-infected HCEn cells were co-cultured with allogeneic CD8+ T cells from CMV seropositive donors and tested for priming activity for the CD8+ effector T cells by measuring interferon-γ secretion. The CMV-induced responses of HCEn cells were characterized by type I interferon and pattern recognition receptor pathways which represent innate immune priming. The global transcriptional activation was specifically associated with antigen presentation with the antimicrobial response functions. Protein array analyses indicated a significant increase in the secretion of anti-viral inflammatory cytokines including CXCL10 as innate immune responses. When HCEn cells were examined to determine whether CMV infection activated anti-viral acquired immunity, CMV-infected HCEn cells directly stimulated the proliferation of CD8+ T cells from CMV-seropositive donors, and pp65 viral epitope induced interferon-γ secretion from the CD8+ T cells. We conclude that CMV-infected HCEn cells induce innate immune priming along with provisions of acquired immune priming of CD8+ effector T cells. This information should help in the development of useful diagnostic procedures and efficacious therapeutic strategy to treat refractory corneal endotheliitis.

Overview of Cytomegalovirus Ocular Diseases: Retinitis, Corneal Endotheliitis, and Iridocyclitis.

Cytomegalovirus (CMV) infection is a significant clinical concern in newborns, immunocompromised patients with acquired immunodeficiency syndrome (AIDS), and patients undergoing immunosuppressive therapy or chemotherapy. CMV infection affects many organs, such as the lungs, digestive organs, the central nerve system, and eyes. In addition, CMV infection sometimes occurs in immunocompetent individuals. CMV ocular diseases includes retinitis, corneal endotheliitis, and iridocyclitis. CMV retinitis often develops in infected newborns and immunocompromised patients. CMV corneal endotheliitis and iridocyclitis sometimes develop in immunocompetent individuals. Systemic infections and CMV ocular diseases often require systemic treatment in addition to topical treatment.

Topical ganciclovir treatment in patients with cytomegalovirus endotheliitis receiving penetrating keratoplasty.

BACKGROUND: To report seven cases diagnosed as cytomegalovirus endotheliitis and treated with topical 2% ganciclovir following penetrating keratoplasty. DESIGN: A retrospectively comparative case series. PARTICIPANTS: A retrospective interventional case series, including seven eyes of seven patients with cytomegalovirus endotheliitis after penetrating keratoplasty. METHODS: Clinical and immunological characteristics were studied in seven penetrating keratoplasty cases with positive quantitative polymerase chain reaction results for cytomegalovirus DNA from aqueous taps and treated with topical 2% ganciclovir. MAIN OUTCOME MEASURES: Clinical features and responses to topical 2% ganciclovir. RESULTS: Seven immunocompetent patients experienced acute anterior inflammation with graft oedema and pigmented keratic precipitates after penetrating keratoplasty. Their immunological profiles showed immunoglobulin G cytomegalovirus (+) and immunoglobulin M cytomegalovirus (-) in all cases. Topical 2% ganciclovir was prescribed every 2 to 3 h daily as induction therapy and every 4 h as long-term maintenance therapy. All cases had undetectable cytomegalovirus DNA after follow-up aqueous taps. Topical 2% ganciclovir preserved endothelium of cytomegalovirus-infected grafts at early stage and also provided a steady anticytomegalovirus environment for further regrafting in failed grafts at late stage. Acute inflammation reactivated in two cases and was suppressible by steroid under topical ganciclovir. No delayed re-epithelialization and any toxicity were observed. To date, no case treated in this way had displayed cytomegalovirus recurrence. CONCLUSIONS: Continuous topical 2% ganciclovir and a topical steroid adjusted by anterior inflammation are suggested after penetrating keratoplasty in all cases with cytomegalovirus endotheliitis to prevent cytomegalovirus recurrence.

Treatment outcome and risk factors for visual loss in Cytomegalovirus endotheliitis.

BACKGROUND: To determine treatment outcome and risk factors for visual loss in Cytomegalovirus (CMV) endotheliitis. METHODS: Retrospective case-note review of all CMV positive endotheliitis patients seen at the Singapore National Eye Center, for demographics, visual acuity (VA), extent of corneal edema, anterior chamber (AC) activity, ocular history, glaucomatous optic neuropathy (GON), and ganciclovir therapy. Outcome measures were VA, corneal edema, and AC activity. RESULTS: Median age at diagnosis of the 19 patients (21 eyes) was 57 years. Median duration of follow up was 37 months. Sixteen eyes received systemic ganciclovir, and four eyes received ganciclovir gel. The AC inflammation resolved in 19 eyes. The corneal edema resolved in eight eyes, but persisted in 12 eyes. One patient resolved spontaneously. Pre-treatment corneal edema exceeding 75%, older age, GON, and previous corneal graft were risk factors for persistent corneal edema post treatment (P = < 0.001, 0.001, 0.02 and 0.02 respectively, Fisher's exact test), and VA worse than 6/60. CONCLUSIONS: Anterior chamber inflammation resolves with ganciclovir therapy, but severe pre-treatment corneal edema, older age, previous corneal graft, and GON are associated with a poor visual outcome.

Multiple parallel-line endotheliitis--a form of herpes simplex keratitis?

PURPOSE: To report a case of multiple parallel-line endotheliitis with a possible link to herpes simplex virus. CASE REPORT: An 89-year-old woman presented with new onset reduced vision in her left eye. Her visual acuity with pinhole was 20/120 with the affected eye and 20/30 in the right. She had a white left eye with significant corneal edema and keratic precipitates. These were arranged in a striking pattern of multiple parallel lines resembling railroad tracks. She responded very well to topical steroid and was found to have positive immunoglobulin G and immunoglobulin M titers for herpes simplex virus. CONCLUSIONS: Multiple parallel-line endotheliitis may represent a mild expression of herpes simplex keratitis. It appears to be exquisitely sensitive to topical steroid on its own suggesting that immune response has a dominant role in its pathogenesis.

[Strengthen the study of several issues on diagnosis and treatment of the corneal herpetic endotheliitis].

Herpes simplex keratitis (HSK), including epithelial keratitis, stromal keratitis, and endotheliitis, is one of the major blinding eye diseases in China. Corneal herpetic endotheliitis has a variety of clinical manifestations, which causes much difficulty for diagnosis and treatment. This paper suggests a uniform classification of corneal herpetic endotheliitis, which helps to the set up and wide application of a standard treatment protocol. As a result, the cure rate would be increased significantly, corneal endothelium loss due to misdiagnosis would be reduced, and finally the blindness rate of HSK would be lowered.

Establishing an Animal Model of Cytomegalovirus Keratouveitis in Rats: Broad Infection of Anterior Segment Tissue by Cytomegalovirus.

Purpose: Considering the difficulty of obtaining adequate biological tissue in clinical practice, we established an animal model of cytomegalovirus (CMV) keratouveitis in rats and investigated the viral infection sites and corresponding imaging and histopathological features. Methods: Subconjunctival injection and topical use of dexamethasone were used to induce ocular immunosuppression in rats followed by intracameral inoculation of murine cytomegalovirus (MCMV). The clinical manifestations, intraocular pressure (IOP) and imaging changes were observed. Infected eyes were further examined by immunofluorescence, light microscopy, and electron microscopy. MCMV RNA was detected by reverse transcription-polymerase chain reaction. Results: Typical keratouveitis occurred in the experimental rats and was characterized by corneal edema, keratic precipitates, and iridocyclitis with increased IOP. Corneal endothelial lesions displayed as "black holes," enlarged intercellular gaps, and high-intensity cellular infiltration by confocal microscopy, consistent with the pathological changes of "ballooning degeneration," endothelial cell detachment, and inflammatory cell infiltration. Mitochondrial edema was the most prominent organelle lesion in endothelial cells. Trabeculitis, mechanical obstruction of Schlemm's canal, and anterior chamber angle stenosis accounted for elevated IOP. Inflammation of the iris and ciliary body tended to transform into a chronic form. Immunofluorescence revealed that corneal endothelial cells, iris cells, trabecular meshwork cells, and monocytes could be infected by MCMV. MCMV RNA was found in the anterior segments after infection. Conclusions: CMV can widely infect anterior segment tissue, including the corneal endothelium, iris, and trabecular meshwork, in vivo, inducing the corresponding clinical manifestations. Corneal endotheliitis and hypertensive anterior uveitis could be the specific stage of anterior segment infection of CMV.

Development of Cytomegalovirus Corneal Endotheliitis During Long-Term Topical Tacrolimus and Steroid Treatment for Chronic Ocular Surface Inflammatory Diseases.

PURPOSE: We report 3 cases of patients with chronic ocular surface inflammatory disease who developed cytomegalovirus (CMV) corneal endotheliitis during immunosuppressant and steroid treatment. PATIENTS AND METHODS: This is a retrospective observational study analyzing the clinical characteristics and outcomes of 3 patients with ocular surface inflammatory diseases (2 with Mooren ulcer and 1 with idiopathic scleritis) who developed CMV corneal endotheliitis. All patients developed CMV corneal endotheliitis between 8 and 14 months of starting steroid and immunosuppressant treatment, including topical 0.1% tacrolimus. Decimal visual acuity, endothelial counts, and intraocular pressure were analyzed. RESULTS: All patients received topical 0.5% ganciclovir after the diagnosis of CMV corneal endotheliitis, which improved endothelial inflammation. However, all patients developed irreversible mydriasis and required additional surgeries, including endothelial keratoplasty, cataract surgery, and glaucoma surgery. At the final follow-up (14-46 months post-CMV corneal endotheliitis onset), fair outcomes were achieved, as demonstrated by a mean decimal best-corrected visual acuity of 0.3 and a well-controlled intraocular pressure. CONCLUSIONS: Topical steroids and immunosuppressants can induce fulminant CMV corneal endotheliitis with cataract progression and irreversible mydriasis. In these cases, early diagnosis and treatment, including topical 0.5% ganciclovir, glaucoma surgery, cataract surgery, and endothelial keratoplasty, are necessary for preserving the patient's vision.

Characteristics of Corneal Endotheliitis among Different Viruses by in Vivo Confocal Microscopy.

Objectives: To explore the cellular morphological characteristics and changes in corneal endotheliitis among different viruses by in vivo confocal microscopy (IVCM).Methods: Corneal confocal images of 44 eyes of 44 patients with HSV, VZV, CMV and EBV corneal endotheliitis were studied retrospectively. Corneal confocal images of 44 normal eyes were used as controls.Results: The pathogens included cytomegalovirus (n = 20), herpes simplex virus (n = 8), varicella zoster virus (n = 10), and Epstein Barr virus (n = 6). There were no differences in the evaluated structures among the different viruses except for the lengths of the subbasal nerves and Langerhans cell densities. Deviations in endothelial cell layers were not significant among different viruses except for owl's eye morphology.Conclusion: ICVM can assist in diagnosing endotheliitis. The results demonstrate that changes in the cornea were not different among the various viruses except for owl's eye morphology, the lengths of the subbasal nerves and Langerhans cell densities.

Cytomegalovirus endotheliitis with recurrent anterior uveitis and secondary glaucoma misdiagnosed as HLA-B27 uveitis.

A 61-year-old male patient presented with decreased vision and recurrent redness in his right eye since the past 4 years. He had been diagnosed elsewhere as HLA-B27 positive anterior uveitis and was on oral methotrexate and topical corticosteroids for recurrent disease. He was on maximal medical therapy for glaucoma. Examination showed prominent inferior corneal oedema with pigmented keratic precipitates and elevated intraocular pressure. He underwent combined trabeculectomy with mitomycin C and cataract surgery. The aqueous sample tested positive for cytomegalovirus. He responded well to oral valganciclovir with resolution of uveitis, the intraocular pressure was well controlled and the corneal oedema resolved completely.

Prevalence and features of keratitis with quantitative polymerase chain reaction positive for cytomegalovirus.

PURPOSE: To assess corneal scrapings and aqueous humor samples analyzed by polymerase chain reaction (PCR) that were positive for cytomegalovirus (CMV) in patients with keratitis of unknown origin and to investigate their clinical manifestations. DESIGN: Retrospective, interventional case series. PARTICIPANTS: Seventy-eight patients with epithelial (n=37), stromal (n=12), or endothelial keratitis (n=29) of unknown origin examined at the Osaka University Medical Hospital. METHODS: Clinical examination and tears, corneal scrapings, and aqueous humor specimens were evaluated by real-time PCR for CMV. MAIN OUTCOME MEASURES: Quantification of CMV DNA at the diagnosis of each type of keratitis with unknown origin and monitoring during the therapeutic course for CMV-positive cases. RESULTS: No cases of epithelial or stromal keratitis had CMV DNA. Seven of 29 corneal endotheliitis cases (24.1%) were positive for CMV. Cytomegalovirus-positive cases of corneal endotheliitis characterized by localized corneal edema and keratic precipitates included 4 patients who had undergone penetrating keratoplasty and were refractory to the treatment for graft rejection and 3 patients with idiopathic endotheliitis. Cytomegalovirus DNA copy numbers were estimated and ranged from 6.3x10(4) to 3.6x10(6)/ml. In all positive cases, the numbers of CMV DNA copies decreased within weeks during treatment with systemic and topical ganciclovir (GCV) combined with a topical steroid. Five eyes (62.5%) had clinical improvement. In cases of endothelial keratitis, diabetes mellitus was significantly higher in patients positive for CMV (71.4%) than in patients negative for CMV (18.2%, P=0.016, chi-square test). CONCLUSIONS: A total of 24.1% of cases with corneal edema of unknown origin were CMV positive and should be included in the differential diagnosis of idiopathic corneal endotheliitis or graft edema after penetrating keratoplasty, especially for bullous keratopathy. Real-time PCR for CMV, based on the diagnosis and monitoring of the clinical course, may be useful. Cytomegalovirus corneal endotheliitis requires early appropriate treatment using GCV. Because clinical remission after GCV may depend on the area of normal endothelium, early diagnosis and therapy are important for CMV corneal endotheliitis.

Recurrent cytomegalovirus corneal endotheliitis after penetrating keratoplasty.

PURPOSE: To report a case of recurrent cytomegalovirus (CMV) corneal endotheliitis after penetrating keratoplasty. METHODS: Penetrating keratoplasty (PK) was performed in a 49-year-old man with bullous keratopathy. Pigmented keratic precipitates (KPs) were found in the corneal endothelium of the graft 13 days after surgery, with the subsequent appearance of coin-shaped lesions. Confocal microscopy was performed on the corneal endothelium. Aqueous humor was analyzed for viral DNA by polymerase chain reaction (PCR). RESULTS: CMV DNA was detected from the excised corneal button, and aqueous humor revealed positive results for CMV and HSV1 by PCR. Confocal microscopy showed large corneal endothelial cells, consistent with the typical owl's eye morphology of CMV endotheliitis. After systemic ganciclovir was administered, the pigmented KPs and coin-shaped lesions gradually decreased. CONCLUSIONS: Stress from surgery and corticosteroid usage can revitalize CMV activity. PCR and confocal microscopy are valuable for the diagnosis of CMV corneal endotheliitis.

Clinical Evaluation of Intravitreal Injection of Ganciclovir in Refractory Corneal Endotheliitis.

Purpose: To evaluate the efficacy and safety of intravitreal ganciclovir (GCV) injection in refractory endotheliitis.Methods: Retrospectively recruited 25 eyes with endotheliitis, proved by clinical manifestations, positive PCR for viral DNA and responded poor to topical and systemic antiviral medications. All patients received additional continued intravitreal GCV injections.Results: Cytomegalovirus (CMV), varicella zoster virus (VZV), and herpes simplex virus (HSV) DNA were detected in 64.0%, 28.0%, and 8.0% of the eyes, respectively. Within 2 weeks after the last injection, 16/25 eyes recovered corneal clarity; active keratic precipitates (KPs) were eliminated in 21/25 eyes; intraocular pressure (IOP) was controlled in 12/15 eyes with elevated IOP on study entry. Best-corrected visual acuity increased at the last follow-up (p = 0.016). Clinical recurrence occurred in three patients. No complications were detected.Conclusions: CMV endotheliitis was the main type of refractory endotheliitis. Despite its invasive nature, intravitreal GCV injection appears to be an effective method for refractory endotheliitis.

Cytomegalovirus Endotheliitis After Penetrating Keratoplasty.

Cytomegalovirus (CMV)-related corneal endotheliitis is an inflammation of the corneal endothelium caused by CMV. It may occur de novo or after ocular surgery in otherwise healthy individuals. In patients who have undergone keratoplasty, the differential diagnosis of viral endotheliitis and immune-related graft rejection is challenging due to the similar clinical findings. Here we report a patient who underwent penetrating keratoplasty and was using local and systemic immunosuppressive agents due to previous history of graft rejection. At postoperative year 4, ophthalmologic examination revealed localized corneal edema, coin-shaped keratic precipitates, and increased intraocular pressure, consistent with viral endotheliitis. Polymerase chain reaction revealed CMV-DNA amplification in the aqueous humor sample. Valganciclovir treatment was started and the symptoms improved in 2 months. It should be kept in mind that local or systemic immunosuppressants used after keratoplasty may trigger CMV reactivation. Anti-CMV treatment should be initiated immediately in patients with coin-shaped keratic precipitates.

Metagenomic Deep Sequencing to Investigate for an Infectious Etiology of Iridocorneal Endothelial Syndrome.

PURPOSE: Iridocorneal endothelial (ICE) syndrome is a group of rare ocular conditions that result from abnormal corneal endothelial cells, leading to secondary glaucoma, iris distortions, and corneal edema. The etiology of ICE is unknown, although it has been associated with viral infections, such as herpes simplex virus. In this study, we sought to identify an infectious etiology for ICE using advanced molecular techniques. METHODS: Metagenomic RNA sequencing (MDS) is a high-throughput sequencing approach that can identify all pathogens in any clinical sample, including RNA viruses. Descemet membrane and aqueous fluid from patients with ICE syndrome were subjected to MDS testing. RESULTS: Samples from 3 patients with ICE were analyzed. MDS was performed on the aqueous fluid of 3 patients and Descemet membrane and endothelial cell tissue from 1 patient. Viral pathogens were not identified in any of the samples. CONCLUSIONS: We were unable to identify a viral etiology in the tissues of patients with the Chandler variant of ICE syndrome, although this study was limited by sample size.

Effects of ROCK Inhibitors on Apoptosis of Corneal Endothelial Cells in CMV-Positive Posner-Schlossman Syndrome Patients.

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.