Critical role of type III interferon in controlling SARS-CoV-2 infection, replication and spread in primary human intestinal epithelial cells
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Abstract
SARS-CoV-2 is an unprecedented worldwide health problem that requires concerted and global approaches to better understand the virus in order to develop novel therapeutic approaches to stop the COVID-19 pandemic and to better prepare against potential future emergence of novel pandemic viruses. Although SARS-CoV-2 primarily targets cells of the lung epithelium causing respiratory infection and pathologies, there is growing evidence that the intestinal epithelium is also infected. However, the importance of the enteric phase of SARS-CoV-2 for virus-induced pathologies, spreading and prognosis remains unknown. Here, using both colon-derived cell lines and primary non-transformed colon organoids, we engage in the first comprehensive analysis of SARS-CoV-2 lifecycle in human intestinal epithelial cells. Our results demonstrate that human intestinal epithelial cells fully support SARS-CoV-2 infection, replication and production of infectious de-novo virus particles. Importantly, we identified intestinal epithelial cells as the best culture model to propagate SARS-CoV-2. We found that viral infection elicited an extremely robust intrinsic immune response where, interestingly, type III interferon mediated response was significantly more efficient at controlling SARS-CoV-2 replication and spread compared to type I interferon. Taken together, our data demonstrate that human intestinal epithelial cells are a productive site of SARS-CoV-2 replication and suggest that the enteric phase of SARS-CoV-2 may participate in the pathologies observed in COVID-19 patients by contributing in increasing patient viremia and by fueling an exacerbated cytokine response.
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SciScore for 10.1101/2020.04.24.059667: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: This study was carried out in accordance with the recommendations of the University Hospital Heidelberg with informed written consent from all subjects in accordance with the Declaration of Helsinki. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibodies and Inhibitors: Mouse monoclonal antibody against SARS-CoV NP (Sino biologicals MM05), mouse monoclonal against J2 (scions), mouse monoclonal against E-cadherin (BD Transductions #610181), and rabbit polyclonal anti-Mucin-2 (Santa Graz Biotechnology# … SciScore for 10.1101/2020.04.24.059667: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: This study was carried out in accordance with the recommendations of the University Hospital Heidelberg with informed written consent from all subjects in accordance with the Declaration of Helsinki. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibodies and Inhibitors: Mouse monoclonal antibody against SARS-CoV NP (Sino biologicals MM05), mouse monoclonal against J2 (scions), mouse monoclonal against E-cadherin (BD Transductions #610181), and rabbit polyclonal anti-Mucin-2 (Santa Graz Biotechnology# sc-15334) were used at 1:500 for immunofluorescence. SARS-CoV NPsuggested: (Sino Biological Cat# 40143-MM05, RRID:AB_2827977)J2suggested: NoneE-cadherinsuggested: (BD Biosciences Cat# 610181, RRID:AB_397580)anti-Mucin-2suggested: (Acris Antibodies GmbH Cat# SM1709, RRID:AB_1619486)Secondary antibody (anti-mouse CW800) and DNA dye Draq5 (Abcam) were diluted 1/10,000 in blocking buffer and incubated for 1h at RT. anti-mouse CW800suggested: NoneDraq5suggested: (Biostatus Cat# DR50050, RRID:AB_2314341)Experimental Models: Cell Lines Sentences Resources Caco-2 human colorectal adenocarcinoma (ATCC HTB-37) and Vero E6 (ATCC CRL 1586) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (GibCo) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (Gibco). Caco-2suggested: NoneIn-cell Western: 20,000 Vero E6 cells were seeded per well into a 96-well dish 24h prior to infection. Vero E6suggested: NoneSoftware and Algorithms Sentences Resources Statistics: Unless otherwise stated, statistical analysis was performed by a two-tailed unpaired t test using the GraphPad Prism software package. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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