In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates

• Published in: BMC ophthalmology 2022-12, Vol.22 (1), p.518-518, Article 518
• Main Authors: Krishna, Venkatramana D, Roehrich, Heidi, Schroeder, Declan C, Cheeran, Maxim C-J, Yuan, Ching, Hou, Joshua H
• Format: Article
• Language: English
• Subjects: Conjunctiva; Cornea - surgery; Corneal endothelium; Corneal epithelium; COVID-19; Disease susceptibility; Endothelium; Endothelium, Corneal - transplantation; Health aspects; Humans; Infection; SARS-CoV-2
• ISSN: 1471-2415; 1471-2415
• DOI: 10.1186/s12886-022-02728-w

The purpose of this study was: [1] to evaluate the infectivity of two SARS-CoV-2 lineage A variants on human ocular tissues in vitro, and [2] to evaluate the stability of SARS-CoV-2 lineage A variants in corneal preservation medium. Primary cultures of donor corneal, conjunctival, and limbal epithelium were inoculated with two lineage A, GISAID clade S isolates of SARS-CoV-2 (Hong Kong/VM20001061/2020, USA-WA1/2020), to evaluate the susceptibility of the ocular tissue to infection. Flat-mounted Descemet's Stripping Automated Endothelial Keratoplasty (DSAEK) grafts were inoculated with SARS-CoV-2 to evaluate the susceptibility of the endothelium to infection. All inoculated samples were immunostained for SARS-CoV-2 nucleocapsid (N)-protein expression to confirm positive infection. SARS-CoV-2 Hong Kong was then inoculated into cornea preservation media (Life4°C, Numedis, Inc.). Inoculated media was stored at 4 C for 14 days and assayed over time for changes in infectious viral titers. Corneal, conjunctival, and limbal epithelial cells all demonstrated susceptibility to infection by SARS-CoV-2 lineage A variants. Conjunctiva demonstrated the highest infection rate (78% of samples infected [14/18]); however, infection rates did not differ statistically between cell types and viral isolates. After inoculation, 40% (4/10) of DSAEK grafts had active infection in the endothelium. SARS-CoV-2 lineage A demonstrated