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Exploration of the ocular surface infection by SARS-CoV-2 and implications for corneal donation: An ex vivo study.
Maurin, Corantin; He, Zhiguo; Mentek, Marielle; Verhoeven, Paul; Pillet, Sylvie; Bourlet, Thomas; Rogues, Françoise; Pugniet, Jean Loup; Peyragrosse, Thierry; Barallon, Marion; Perrache, Chantal; Aouimeur, Inès; Acquart, Sophie; Ninotta, Sandrine; Baud'huin, Marc; Vabres, Bertrand; Poinard, Sylvain; Gain, Philippe; Thuret, Gilles.
Afiliação
  • Maurin C; Laboratory "Biology, engineering and imaging of Corneal Graft" BiiGC, Faculty of Medicine, University Jean Monnet, Saint-Etienne, France.
  • He Z; Laboratory "Biology, engineering and imaging of Corneal Graft" BiiGC, Faculty of Medicine, University Jean Monnet, Saint-Etienne, France.
  • Mentek M; Laboratory "Biology, engineering and imaging of Corneal Graft" BiiGC, Faculty of Medicine, University Jean Monnet, Saint-Etienne, France.
  • Verhoeven P; CIRI, Centre International de Recherche en Infectiologie, GIMAP Team University of Lyon, University of St-Etienne, INSERM U1111, CNRS UMR5308, ENS de Lyon, UCBL1, St-Etienne, France.
  • Pillet S; Laboratory of Infectious Agents and Hygiene, University Hospital of St-Etienne, St-Etienne, France.
  • Bourlet T; CIRI, Centre International de Recherche en Infectiologie, GIMAP Team University of Lyon, University of St-Etienne, INSERM U1111, CNRS UMR5308, ENS de Lyon, UCBL1, St-Etienne, France.
  • Rogues F; Laboratory of Infectious Agents and Hygiene, University Hospital of St-Etienne, St-Etienne, France.
  • Pugniet JL; CIRI, Centre International de Recherche en Infectiologie, GIMAP Team University of Lyon, University of St-Etienne, INSERM U1111, CNRS UMR5308, ENS de Lyon, UCBL1, St-Etienne, France.
  • Peyragrosse T; Laboratory of Infectious Agents and Hygiene, University Hospital of St-Etienne, St-Etienne, France.
  • Barallon M; Hospital coordination of organ and/or tissue retrieval, University Hospital, Saint-Etienne, France.
  • Perrache C; Hospital coordination of organ and/or tissue retrieval, University Hospital, Saint-Etienne, France.
  • Aouimeur I; Hospital coordination of organ and/or tissue retrieval, University Hospital, Saint-Etienne, France.
  • Acquart S; Hospital coordination of organ and/or tissue retrieval, University Hospital, Saint-Etienne, France.
  • Ninotta S; Laboratory "Biology, engineering and imaging of Corneal Graft" BiiGC, Faculty of Medicine, University Jean Monnet, Saint-Etienne, France.
  • Baud'huin M; Laboratory "Biology, engineering and imaging of Corneal Graft" BiiGC, Faculty of Medicine, University Jean Monnet, Saint-Etienne, France.
  • Vabres B; Eye bank, French Blood Center, Saint-Etienne, France.
  • Poinard S; Eye bank, French Blood Center, Saint-Etienne, France.
  • Gain P; Nantes Université, CHU Nantes, Banque Muti-Tissus, Nantes, France.
  • Thuret G; Nantes Université, CHU Nantes, Banque Muti-Tissus, Nantes, France.
PLoS Med ; 19(3): e1003922, 2022 03.
Article em En | MEDLINE | ID: mdl-35231027
BACKGROUND: The risk of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) transmission through corneal graft is an ongoing debate and leads to strict restrictions in corneas procurement, leading to a major decrease in eye banking activity. The aims of this study are to specifically assess the capacity of human cornea to be infected by SARS-CoV-2 and promote its replication ex vivo, and to evaluate the real-life risk of corneal contamination by detecting SARS-CoV-2 RNA in corneas retrieved in donors diagnosed with Coronavirus Disease 2019 (COVID-19) and nonaffected donors. METHODS AND FINDINGS: To assess the capacity of human cornea to be infected by SARS-CoV-2, the expression pattern of SARS-CoV-2 receptor angiotensin-converting enzyme 2 (ACE-2) and activators TMPRSS2 and Cathepsins B and L in ocular surface tissues from nonaffected donors was explored by immunohistochemistry (n = 10 corneas, 78 ± 11 years, 40% female) and qPCR (n = 5 corneas, 80 ± 12 years, 40% female). Additionally, 5 freshly excised corneas (80 ± 12 years, 40% female) were infected ex vivo with highly concentrated SARS-CoV-2 solution (106 median tissue culture infectious dose (TCID50)/mL). Viral RNA was extracted from tissues and culture media and quantified by reverse transcription quantitative PCR (RT-qPCR) (viral RNA copies) 30 minutes (H0) and 24 hours (H24) after infection. To assess the risk of corneal contamination by SARS-CoV-2, viral RNA was tested by RT-qPCR (Ct value) in both corneas and organ culture media from 14 donors diagnosed with COVID-19 (74 ± 10 years, 29% female) and 26 healthy donors (79 ± 13 years, 57% female), and in organ culture media only from 133 consecutive nonaffected donors from 2 eye banks (73 ± 13 years, 29% female). The expression of receptor and activators was variable among samples at both protein and mRNA level. Based on immunohistochemistry findings, ACE-2 was localized mainly in the most superficial epithelial cells of peripheral cornea, limbus, and conjunctiva, whereas TMPRSS2 was mostly expressed in all layers of bulbar conjunctiva. A significant increase in total and positive strands of IP4 RNA sequence (RdRp viral gene) was observed from 30 minutes to 24 hours postinfection in central cornea (1.1 × 108 [95% CI: 6.4 × 107 to 2.4 × 108] to 3.0 × 109 [1.4 × 109 to 5.3 × 109], p = 0.0039 and 2.2 × 107 [1.4 × 107 to 3.6 × 107] to 5.1 × 107 [2.9 × 107 to 7.5 × 107], p = 0.0117, respectively) and in corneoscleral rim (4.5 × 109 [2.7 × 109 to 9.6 × 109] to 3.9 × 1010 [2.6 × 1010 to 4.4 × 1010], p = 0.0039 and 3.1 × 108 [1.2 × 108 to 5.3 × 108] to 7.8 × 108 [3.9 × 108 to 9.9 × 108], p = 0.0391, respectively). Viral RNA copies in ex vivo corneas were highly variable from one donor to another. Finally, viral RNA was detected in 3 out of 28 corneas (11%) from donors diagnosed with COVID-19. All samples from the 159 nonaffected donors were negative for SARS-CoV-2 RNA. The main limitation of this study relates to the limited sample size, due to limited access to donors diagnosed with COVID-19 and concomitant decrease in the procurement corneas from nonaffected donors. CONCLUSIONS: In this study, we observed the expression of SARS-CoV-2 receptors and activators at the human ocular surface and a variable increase in viral RNA copies 24 hours after experimental infection of freshly excised human corneas. We also found viral RNA only in a very limited percentage of donors with positive nasopharyngeal PCR. The low rate of positivity in donors diagnosed with COVID-19 calls into question the utility of donor selection algorithms. TRIAL REGISTRATION: Agence de la Biomédecine, PFS-20-011 https://www.agence-biomedecine.fr/.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Infecções Oculares Virais / Córnea / Doenças da Córnea / SARS-CoV-2 / COVID-19 Limite: Adult / Aged / Animals / Female / Humans / Male / Middle aged Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Infecções Oculares Virais / Córnea / Doenças da Córnea / SARS-CoV-2 / COVID-19 Limite: Adult / Aged / Animals / Female / Humans / Male / Middle aged Idioma: En Ano de publicação: 2022 Tipo de documento: Article