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

Venkatramana D Krishna; Heidi Roehrich; Declan C Schroeder; Maxim C-J Cheeran; Ching Yuan; Joshua H Hou

doi:10.1186/s12886-022-02728-w

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

BMC Ophthalmol. 2022 Dec 30;22(1):518. doi: 10.1186/s12886-022-02728-w.

Authors

Venkatramana D Krishna¹, Heidi Roehrich², Declan C Schroeder^{1

3}, Maxim C-J Cheeran¹, Ching Yuan^{2

4}, Joshua H Hou^{5

6}

Affiliations

¹ Department of Veterinary Population Medicine, University of Minnesota, Minneapolis, MN, USA.
² Department of Ophthalmology and Visual Neurosciences, University of Minnesota, 420 Delaware St SE, MMC 493, Minneapolis, MN, 55455, USA.
³ School of Biological Sciences, University of Reading, Reading, UK.
⁴ Lions Gift of Sight Eye Bank, St. Paul, MN, USA.
⁵ Department of Ophthalmology and Visual Neurosciences, University of Minnesota, 420 Delaware St SE, MMC 493, Minneapolis, MN, 55455, USA. jhhou@umn.edu.
⁶ Lions Gift of Sight Eye Bank, St. Paul, MN, USA. jhhou@umn.edu.

Abstract

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 variants 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 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^oC for 14 days and assayed over time for changes in infectious viral titers.

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.

Keywords: COVID-19; Corneal endothelium; Corneal epithelium; SARS-CoV-2.

MeSH terms

COVID-19*
Conjunctiva
Cornea / surgery
Endothelium, Corneal / transplantation
Humans
SARS-CoV-2*