Varicella zoster virus-induced autophagy in human neuronal and hematopoietic cells exerts antiviral activity.

Autophagy is a degradational pathway with pivotal roles in cellular homeostasis and survival, including protection of neurons in the central nervous system (CNS). The significance of autophagy as antiviral defense mechanism is recognized and some viruses hijack and modulate this process to their advantage in certain cell types. Here, we present data demonstrating that the human neurotropic herpesvirus varicella zoster virus (VZV) induces autophagy in human SH-SY5Y neuronal cells, in which the pathway exerts antiviral activity. Productively VZV-infected SH-SY5Y cells showed increased LC3-I-LC3-II conversion as well as co-localization of the viral glycoprotein E and the autophagy receptor p62. The activation of autophagy was dependent on a functional viral genome. Interestingly, inducers of autophagy reduced viral transcription, whereas inhibition of autophagy increased viral transcript expression. Finally, the genotype of patients with severe ocular and brain VZV infection were analyzed to identify potential autophagy-associated inborn errors of immunity. Two patients expressing genetic variants in the autophagy genes ULK1 and MAP1LC3B2, respectively, were identified. Notably, cells of both patients showed reduced autophagy, alongside enhanced viral replication and death of VZV-infected cells. In conclusion, these results demonstrate a neuro-protective role for autophagy in the context of VZV infection and suggest that failure to mount an autophagy response is a potential predisposing factor for development of severe VZV disease.

Risk Factors for Cytomegalovirus Retinitis in a National Survey in Sweden.

PURPOSE: To investigate predisposing immunosuppressive conditions causing cytomegalovirus retinitis (CMVr) and risk factors for delayed diagnosis in patients diagnosed between 2008 and 2018 in the Swedish population of 10 million. RESULTS: Sixty-three consecutive patients (100 eyes) were diagnosed with CMVr. The most common immunosuppressive state predisposing for CMVr was hematopoietic stem cell transplant recipients (27%) and hematological malignancy (24%). Two patients (3.2%) had no other predisposing factor than diabetes mellitus (DM). Patients with delayed diagnosis (≤30 days) were older than those with earlier diagnosis, mean age 68.7 (±9.8) and 48.8 (±17.6), respectively, p = .001. Signs of intraocular inflammation (IOI) were seen in 42 (70%) of cases and more common in delayed than early diagnosis, 16 (89%) and 21 (60%) respectively (p = .03). CONCLUSION: Delayed diagnosis was more common in older individuals with signs of IOI. DM may be a risk factor for CMVr, and clinical signs can be mistaken for diabetic retinopathy.

Whole exome sequencing of patients with varicella-zoster virus and herpes simplex virus induced acute retinal necrosis reveals rare disease-associated genetic variants.

Purpose: Herpes simplex virus (HSV) and varicella-zoster virus (VZV) are neurotropic human alphaherpesviruses endemic worldwide. Upon primary infection, both viruses establish lifelong latency in neurons and reactivate intermittently to cause a variety of mild to severe diseases. Acute retinal necrosis (ARN) is a rare, sight-threatening eye disease induced by ocular VZV or HSV infection. The virus and host factors involved in ARN pathogenesis remain incompletely described. We hypothesize an underlying genetic defect in at least part of ARN cases. Methods: We collected blood from 17 patients with HSV-or VZV-induced ARN, isolated DNA and performed Whole Exome Sequencing by Illumina followed by analysis in Varseq with criteria of CADD score > 15 and frequency in GnomAD < 0.1% combined with biological filters. Gene modifications relative to healthy control genomes were filtered according to high quality and read-depth, low frequency, high deleteriousness predictions and biological relevance. Results: We identified a total of 50 potentially disease-causing genetic variants, including missense, frameshift and splice site variants and on in-frame deletion in 16 of the 17 patients. The vast majority of these genes are involved in innate immunity, followed by adaptive immunity, autophagy, and apoptosis; in several instances variants within a given gene or pathway was identified in several patients. Discussion: We propose that the identified variants may contribute to insufficient viral control and increased necrosis ocular disease presentation in the patients and serve as a knowledge base and starting point for the development of improved diagnostic, prophylactic, and therapeutic applications.

Absence of Herpesvirus DNA in Aqueous Humor from Asymptomatic Subjects.

Purpose: To assess herpesvirus DNA detection in aqueous humor from a cohort of asymptomatic Scandinavian patients undergoing elective cataract surgery. Patients and Methods: Prospective case series. Aqueous samples were obtained from 30 patients undergoing elective cataract surgery. Polymerase chain reaction (PCR) analysis for herpes simplex virus 1 (HSV1), herpes simplex virus 2 (HSV2), cytomegalovirus (CMV), Epstein Barr virus (EBV) was performed. Toxoplasma was added to the analysis due to its role as pathogen with ocular latency. Results: Mean age of participants was 75.3 years. Sixteen subjects (53%) had ocular comorbidities. Five subjects (17%) had endothelial dysfunction without known hereditary pattern. None of the samples were positive for herpesviruses or toxoplasma. Conclusion: None of the aqueous samples were positive, suggesting shedding does not frequently occur in the aqueous humor of asymptomatic patients.

Deep Sequencing of Varicella-Zoster Virus in Aqueous Humor From a Patient With Acute Retinal Necrosis Presenting With Acute Glaucoma.

We report a case of acute retinal necrosis presenting with acute glaucoma preceding inflammatory signs by several days. High-throughput sequencing on aqueous humor revealed a low-level diversity in the viral genome comparable to diversity seen in cutaneous vesicles in contrast to high diversity in encephalitis.

Alpha herpes virus type and viral load in intraocular fluids in patients with acute retinal necrosis.

OBJECTIVES: To identify all patients tested positive for herpes viruses in intraocular samples between 2007 and 2016 in South-Western Sweden and evaluate which of these met the criteria of acute retinal necrosis (ARN). To compare viral load in intraocular samples and virus type with clinical outcome. METHOD AND ANALYSIS: Retrospective case series. Intraocular samples and serum were analysed with quantitative real-time PCR (qPCR) and presence of antibodies (IgG and IgM) were detected by ELISA in serum. RESULTS: Between 2007 and 2016, 13 patients met the clinical criteria of ARN and were PCR-positive in aqueous or vitreous for herpes simplex virus 1 (HSV1; n=4), herpes simplex virus 2 (HSV2; n=3) and varicella zoster virus (VZV; n=6). None of the patients tested positive for cytomegalovirus (n=13) or Epstein Barr virus (n=2) met the criteria of ARN. All ARN patients had specific serum IgG and three patients exhibited virus DNA in serum. There was no correlation between high viral load and worse visual outcome. However, higher viral loads were seen in samples taken earlier in the disease process. Median age was higher (p=0.049) in VZV-ARN than for HSV-ARN patients (60.5 and 45.4 years, respectively) with a tendency of worse best corrected visual acuity at presentation (1.62 and 0.79 log MAR, respectively; p=0.079). CONCLUSION: ARN is a reactivation of alpha herpes virus and presence of herpes DNA in serum may occur. VZV-ARN are older than HSV-ARN patients. High viral load does not appear to be a predictor of worse visual outcome, but rather indicates earlier sampling.

The use of autologous serum for the treatment of ocular surface disease at a Swedish tertiary referral center.

PURPOSE: The study aims to describe an intact cohort with mixed ocular surface disease (OSD) treated with autologous serum (AS) eye drops in a tertiary eye center. PATIENTS AND METHODS: All cases (n=32 eyes, 24 patients) treated with AS for OSD at the Department of Ophthalmology, Sahlgrenska University Hospital, Mölndal, between 2002 and 2013 were included and medical records were reviewed retrospectively. RESULTS: Mean duration of treatment with 20% AS was 28.3±56.1 (median: 12, range: 3-217) days. The most common indication for AS treatment was a persistent epithelial defect (PED), which was seen in 16 eyes of 14 patients. Mean duration of PED prior to treatment was 19.3±18.9 (median: 10, range: 5-68) days. Complete or partial epithelial healing occurred in nine eyes (56.2%). The remaining seven eyes (44%) did not respond to treatment or data were missing. The second group consisted of nine eyes of five patients with superficial punctate keratitis (SPK) secondary to dry eye syndrome. Complete or partial healing of the epithelium occurred in five eyes (56%), and the remaining four eyes (44%) were lost to follow-up. A third group included five eyes with AS as an adjuvant treatment after corneal perforation, whereas a fourth group consisted of one patient with dry eye after laser-assisted in situ keratomileusis (LASIK). CONCLUSION: In this cohort, patients with PED or SPK responded well to treatment with AS. Standardized preparation protocols, defined optimal serum concentrations for various indications, and large randomized clinical trials are needed to fully comprehend the role of AS in the treatment of OSD.