In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates
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, conj...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-12-01
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| Series: | BMC Ophthalmology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12886-022-02728-w |
| _version_ | 1797973703413202944 |
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| author | Venkatramana D. Krishna Heidi Roehrich Declan C. Schroeder Maxim C.-J. Cheeran Ching Yuan Joshua H. Hou |
| author_facet | Venkatramana D. Krishna Heidi Roehrich Declan C. Schroeder Maxim C.-J. Cheeran Ching Yuan Joshua H. Hou |
| author_sort | Venkatramana D. Krishna |
| collection | DOAJ |
| description | 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 4oC 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. |
| first_indexed | 2024-04-11T04:08:33Z |
| format | Article |
| id | doaj.art-711dc278b83f4478bb597d62835a58b1 |
| institution | Directory Open Access Journal |
| issn | 1471-2415 |
| language | English |
| last_indexed | 2024-04-11T04:08:33Z |
| publishDate | 2022-12-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Ophthalmology |
| spelling | doaj.art-711dc278b83f4478bb597d62835a58b12023-01-01T12:15:32ZengBMCBMC Ophthalmology1471-24152022-12-012211910.1186/s12886-022-02728-wIn vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolatesVenkatramana D. Krishna0Heidi Roehrich1Declan C. Schroeder2Maxim C.-J. Cheeran3Ching Yuan4Joshua H. Hou5Department of Veterinary Population Medicine, University of MinnesotaDepartment of Ophthalmology and Visual Neurosciences, University of MinnesotaDepartment of Veterinary Population Medicine, University of MinnesotaDepartment of Veterinary Population Medicine, University of MinnesotaDepartment of Ophthalmology and Visual Neurosciences, University of MinnesotaDepartment of Ophthalmology and Visual Neurosciences, University of MinnesotaAbstract 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 4oC 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.https://doi.org/10.1186/s12886-022-02728-wSARS-CoV-2COVID-19Corneal epitheliumCorneal endothelium |
| spellingShingle | Venkatramana D. Krishna Heidi Roehrich Declan C. Schroeder Maxim C.-J. Cheeran Ching Yuan Joshua H. Hou In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates BMC Ophthalmology SARS-CoV-2 COVID-19 Corneal epithelium Corneal endothelium |
| title | In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates |
| title_full | In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates |
| title_fullStr | In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates |
| title_full_unstemmed | In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates |
| title_short | In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates |
| title_sort | in vitro infection of human ocular tissues by sars cov 2 lineage a isolates |
| topic | SARS-CoV-2 COVID-19 Corneal epithelium Corneal endothelium |
| url | https://doi.org/10.1186/s12886-022-02728-w |
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