Absence of detectable SARS-CoV-2 replication in ex vivo cultured cornea and cornea-derived epithelial cells
© 2022. The Author(s)..
PURPOSE: To study the possibility of SARS-CoV-2 to infect human corneal cells and tissues under standard corneal culture conditions using explants of COVID-19 donors and primary cornea-derived epithelial cells.
METHODS: Cornea isolated from deceased COVID-19 donors was cultured for 4 weeks, and SARS-CoV-2 replication was monitored by qRT-PCR. Furthermore, primary corneal epithelial cells from healthy donors were cultured ex vivo and infected with SARS-CoV-2 and human cytomegalovirus (HCMV) as a control. Infection status was assessed by western blotting and reporter gene expression using green fluorescent protein-expressing viral strains. ACE2 and TMPRSS2 receptor expression levels in cornea and epithelial cells were assessed by qRT-PCR.
RESULTS: We did not detect SARS-CoV-2 replication in 10 corneas isolated from deceased COVID-19 patients and cultured for 4 weeks, indicating absence of infection under natural conditions. Furthermore, high-titer SARS-CoV-2 infection of ex vivo cultured cornea-derived epithelial cells did not result in productive virus replication. In contrast, the same cells were highly permissive for HCMV. This phenotype could potentially be explained by low ACE2 and TMPRSS2 transcriptional activity in cornea and cornea-derived epithelial cells.
CONCLUSIONS: Our data suggest that cornea and limbal epithelial cells are refractory to productive SARS-CoV-2 infection. This could be due to the absence of robust receptor expression levels necessary for viral entry. This study adds further evidence to support the very low possibility of transmission of SARS-CoV-2 from an infected corneal transplant donor to a recipient in corneal organ cultures.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:261 |
|---|---|
| Enthalten in: |
Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie - 261(2023), 2 vom: 16. Feb., Seite 435-446 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Bayyoud, Tarek [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 23.01.2023 Date Revised 23.01.2023 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1007/s00417-022-05776-6 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM344404714 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM344404714 | ||
| 003 | DE-627 | ||
| 005 | 20231226022706.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1007/s00417-022-05776-6 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1147.xml |
| 035 | |a (DE-627)NLM344404714 | ||
| 035 | |a (NLM)35920896 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Bayyoud, Tarek |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Absence of detectable SARS-CoV-2 replication in ex vivo cultured cornea and cornea-derived epithelial cells |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 23.01.2023 | ||
| 500 | |a Date Revised 23.01.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2022. The Author(s). | ||
| 520 | |a PURPOSE: To study the possibility of SARS-CoV-2 to infect human corneal cells and tissues under standard corneal culture conditions using explants of COVID-19 donors and primary cornea-derived epithelial cells | ||
| 520 | |a METHODS: Cornea isolated from deceased COVID-19 donors was cultured for 4 weeks, and SARS-CoV-2 replication was monitored by qRT-PCR. Furthermore, primary corneal epithelial cells from healthy donors were cultured ex vivo and infected with SARS-CoV-2 and human cytomegalovirus (HCMV) as a control. Infection status was assessed by western blotting and reporter gene expression using green fluorescent protein-expressing viral strains. ACE2 and TMPRSS2 receptor expression levels in cornea and epithelial cells were assessed by qRT-PCR | ||
| 520 | |a RESULTS: We did not detect SARS-CoV-2 replication in 10 corneas isolated from deceased COVID-19 patients and cultured for 4 weeks, indicating absence of infection under natural conditions. Furthermore, high-titer SARS-CoV-2 infection of ex vivo cultured cornea-derived epithelial cells did not result in productive virus replication. In contrast, the same cells were highly permissive for HCMV. This phenotype could potentially be explained by low ACE2 and TMPRSS2 transcriptional activity in cornea and cornea-derived epithelial cells | ||
| 520 | |a CONCLUSIONS: Our data suggest that cornea and limbal epithelial cells are refractory to productive SARS-CoV-2 infection. This could be due to the absence of robust receptor expression levels necessary for viral entry. This study adds further evidence to support the very low possibility of transmission of SARS-CoV-2 from an infected corneal transplant donor to a recipient in corneal organ cultures | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a COVID-19 | |
| 650 | 4 | |a Cornea organ transplantation | |
| 650 | 4 | |a HCMV | |
| 650 | 4 | |a Human cytomegalovirus | |
| 650 | 4 | |a Ocular surface | |
| 650 | 4 | |a SARS-CoV-2 | |
| 650 | 7 | |a Angiotensin-Converting Enzyme 2 |2 NLM | |
| 650 | 7 | |a EC 3.4.17.23 |2 NLM | |
| 650 | 7 | |a TMPRSS2 protein, human |2 NLM | |
| 650 | 7 | |a EC 3.4.21.- |2 NLM | |
| 650 | 7 | |a ACE2 protein, human |2 NLM | |
| 650 | 7 | |a EC 3.4.17.23 |2 NLM | |
| 700 | 1 | |a Vavouras Syrigos, Georgios |e verfasserin |4 aut | |
| 700 | 1 | |a Ruetalo Buschinger, Natalia |e verfasserin |4 aut | |
| 700 | 1 | |a Wude, Johanna |e verfasserin |4 aut | |
| 700 | 1 | |a Businger, Ramona |e verfasserin |4 aut | |
| 700 | 1 | |a Hu, Dan |e verfasserin |4 aut | |
| 700 | 1 | |a Iftner, Angelika |e verfasserin |4 aut | |
| 700 | 1 | |a Thaler, Sebastian |e verfasserin |4 aut | |
| 700 | 1 | |a Schindler, Michael |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie |d 1985 |g 261(2023), 2 vom: 16. Feb., Seite 435-446 |w (DE-627)NLM01296803X |x 1435-702X |7 nnns |
| 773 | 1 | 8 | |g volume:261 |g year:2023 |g number:2 |g day:16 |g month:02 |g pages:435-446 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1007/s00417-022-05776-6 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 261 |j 2023 |e 2 |b 16 |c 02 |h 435-446 | ||