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

Background: 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 variant in corneal preservation medium. Methods 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 surface to infection. Cultured human corneal endothelium and flat-mounted 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 o C, Numedis, Inc.). Inoculated media was stored at 4 o C for 14 days and assayed over time for changes in infectious viral titers (TCID 50 ). Results 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 < 1 log decline in viral titers out to 14 days in corneal preservation media. Conclusions SARS-CoV-2 lineage A variants can infect corneal, limbal, and conjunctival epithelium, as well as corneal endothelium. There was no statistical difference in infectivity between different lineage A variants. SARS-CoV-2 lineage A can survive and remain infectious in corneal preservation media out to 14 days in cold storage.