LSC - 2021 - Use of differentiated primary human airway epithelial models to study SARS-CoV-2 infection biology and pathogenesis
W. Ying, M. Thaler (M.Thaler@lumc.nl / Leiden University Medical Center,Leiden), D. K. Ninaber (D.K.Ninaber@lumc.nl / Leiden University Medical Center,Leiden), A. M. Van Der Does (A.van_der_Does@lumc.nl / Leiden University Medical Center,Leiden), N. S. Ogando (N.S.Lima_Leite_Ogando@lumc.nl / Leiden University Medical Center,Leiden), H. Beckert (hendrik.beckert@rlk.uk-essen.de / University Medical Center Essen – Ruhrlandklinik,Essen), C. Taube (christian.taube@rlk.uk-essen.de / University Medical Center Essen – Ruhrlandklinik,Essen), C. Salgado Benvindo Da Silva (C.Salgado_Benvindo_da_Silva@lumc.nl / Leiden University Medical Center,Leiden), P. J. Bredenbeek (p.j.bredenbeek@lumc.nl / Leiden University Medical Center,Leiden), P. S. Hiemstra (P.S.Hiemstra@lumc.nl / Leiden University Medical Center,Leiden), M. J. Van Hemert (M.J.van_Hemert@lumc.nl / Leiden University Medical Center,Leiden)
Source: Virtual Congress 2021 – Emerging respiratory diseases: state-of-the-art studies of SARS-CoV-2 infection
Abstract The coronavirus disease 2019 (COVID-19) global pandemic, which so far has resulted in about 1 million deaths, is caused by severe acute respiratory coronavirus 2 (SARS-CoV-2). At present, infection dynamics and pathogenesis of the disease are incompletely understood. Here, we used well-differentiated primary human tracheal and bronchial epithelial cells (PTEC/PBEC) cultured at the air-liquid interface (ALI) to study the infection kinetics and subsequent epithelial response. ALI-PTEC and PBEC were differentiated for 3, 4 or 5 weeks, and next infected with SARS-CoV-2 to assess the impact of differentiation status on replication dynamics. We observed a gradual increase in viral load with prolonged differentiation time, and therefore focused on 5-weeks differentiated cultures. Next, we assessed the epithelial response to infection, identified the epithelial cell types that are predominantly infected and studied expression of SARS-CoV-2 entry-related host factors by using various methods (PCR, viral plaque assay and immunofluorescence [IF]). We found that ALI-PTEC cultures contained more goblet cells and less ciliated cells and showed higher expression of angiotensin-converting enzyme 2 (ACE2) compared to ALI-PBEC. Despite these differences, both cultures were efficiently infected by SARS-CoV-2. IF analysis showed that SARS-CoV-2 targets both ciliated and goblet cells. Expression of SARS-CoV-2 entry-related factors (ACE2 and TMPRSS2), IL-8 and IL-6 as well as type I and III interferons were increased after infection. These findings provide a foundation to further study virus-airway epithelial cell interactions in relation to host susceptibility to COVID-19.
This study was supported by a grant from ZonMw and Proefdiervrij (# 114025007).
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W. Ying, M. Thaler (M.Thaler@lumc.nl / Leiden University Medical Center,Leiden), D. K. Ninaber (D.K.Ninaber@lumc.nl / Leiden University Medical Center,Leiden), A. M. Van Der Does (A.van_der_Does@lumc.nl / Leiden University Medical Center,Leiden), N. S. Ogando (N.S.Lima_Leite_Ogando@lumc.nl / Leiden University Medical Center,Leiden), H. Beckert (hendrik.beckert@rlk.uk-essen.de / University Medical Center Essen – Ruhrlandklinik,Essen), C. Taube (christian.taube@rlk.uk-essen.de / University Medical Center Essen – Ruhrlandklinik,Essen), C. Salgado Benvindo Da Silva (C.Salgado_Benvindo_da_Silva@lumc.nl / Leiden University Medical Center,Leiden), P. J. Bredenbeek (p.j.bredenbeek@lumc.nl / Leiden University Medical Center,Leiden), P. S. Hiemstra (P.S.Hiemstra@lumc.nl / Leiden University Medical Center,Leiden), M. J. Van Hemert (M.J.van_Hemert@lumc.nl / Leiden University Medical Center,Leiden). LSC - 2021 - Use of differentiated primary human airway epithelial models to study SARS-CoV-2 infection biology and pathogenesis. 4311
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