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 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 (Life4oC, Numedis, Inc.). Inoculated media was stored at 4oC for 14 days and assayed over time for changes in infectious viral titers (TCID50). 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..
| Medienart: |
|
|---|
Preprint| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
ResearchSquare.com - (2023) vom: 16. Okt. Zur Gesamtaufnahme - year:2023 |
|---|
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Krishna, Venkatramana D. [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
|---|
| doi: |
10.21203/rs.3.rs-1735962/v1 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
XRA03675496X |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | XRA03675496X | ||
| 003 | DE-627 | ||
| 005 | 20231017132543.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 220803s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.21203/rs.3.rs-1735962/v1 |2 doi | |
| 035 | |a (DE-627)XRA03675496X | ||
| 035 | |a (ResearchSquare)10.21203/rs.3.rs-1735962/v1 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Krishna, Venkatramana D. |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a In vitro infection of human ocular tissues by SARS-CoV-2 lineage A isolates |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 520 | |a 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 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 (Life4oC, Numedis, Inc.). Inoculated media was stored at 4oC for 14 days and assayed over time for changes in infectious viral titers (TCID50). 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. | ||
| 650 | 4 | |a Biology |7 (dpeaa)DE-84 | |
| 650 | 4 | |a 570 |7 (dpeaa)DE-84 | |
| 700 | 1 | |a Roehrich, Heidi |4 aut | |
| 700 | 1 | |a Schroeder, Declan C. |4 aut | |
| 700 | 1 | |a Cheeran, Maxim C-J. |4 aut | |
| 700 | 1 | |a Yuan, Ching |4 aut | |
| 700 | 1 | |a Hou, Joshua H |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t ResearchSquare.com |g (2023) vom: 16. Okt. |
| 773 | 1 | 8 | |g year:2023 |g day:16 |g month:10 |
| 856 | 4 | 0 | |u https://doi.org/10.1186/s12886-022-02728-w |z lizenzpflichtig |3 Volltext |
| 856 | 4 | 0 | |u http://dx.doi.org/10.21203/rs.3.rs-1735962/v1 |z kostenfrei |3 Volltext |
| 912 | |a GBV_XRA | ||
| 951 | |a AR | ||
| 952 | |j 2023 |b 16 |c 10 | ||