Immediate myeloid depot for SARS-CoV-2 in the human lung
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Abstract
In the pathogenesis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, epithelial populations in the distal lung expressing Angiotensin-converting enzyme 2 (ACE2) are infrequent, and therefore, the model of viral expansion and immune cell engagement remains incompletely understood. Using human lungs to investigate early host-viral pathogenesis, we found that SARS-CoV-2 had a rapid and specific tropism for myeloid populations. Human alveolar macrophages (AMs) reliably expressed ACE2 allowing both spike-ACE2–dependent viral entry and infection. In contrast to Influenza A virus, SARS-CoV-2 infection of AMs was productive, amplifying viral titers. While AMs generated new viruses, the interferon responses to SARS-CoV-2 were muted, hiding the viral dissemination from specific antiviral immune responses. The reliable and veiled viral depot in myeloid cells in the very early phases of SARS-CoV-2 infection of human lungs enables viral expansion in the distal lung and potentially licenses subsequent immune pathologies.
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SciScore for 10.1101/2022.04.28.489942: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: , Zuckerberg San Francisco General Hospital) under research protocol 20-30497 approved by the University of California San Francisco Institutional Review Board. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources For selected experiments, BAL cells were treated for 2h before infection with an ACE2 blocking antibody at 10 µg/ml (AF933, R&D systems). ACE2suggested: NoneExperimental Models: Cell Lines Sentences Resources SARS-CoV-2 infections: Vero E6 and Vero-TMPRSS2 cells (gift from Dr. Melanie Ott) were cultured in DMEM supplemented with 10% FBS, … SciScore for 10.1101/2022.04.28.489942: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: , Zuckerberg San Francisco General Hospital) under research protocol 20-30497 approved by the University of California San Francisco Institutional Review Board. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources For selected experiments, BAL cells were treated for 2h before infection with an ACE2 blocking antibody at 10 µg/ml (AF933, R&D systems). ACE2suggested: NoneExperimental Models: Cell Lines Sentences Resources SARS-CoV-2 infections: Vero E6 and Vero-TMPRSS2 cells (gift from Dr. Melanie Ott) were cultured in DMEM supplemented with 10% FBS, penicillin/streptomycin, and L-glutamine (Corning) in a humidified incubator at 37°C and 5% CO2. Vero E6suggested: NoneVero-TMPRSS2suggested: JCRB Cat# JCRB1818, RRID:CVCL_YQ48)Briefly, 10-fold dilutions of the virus stock were added to Vero cells in a 12-well plate for 1 hour, after which an overlay of 1.2% Avicel RC-581 in DMEM was added. Verosuggested: NoneThis solution was used as inoculum for Vero E6 cells (SARS-CoV-2) or MDCK cells (IAV-Venus). MDCKsuggested: CLS Cat# 602280/p823_MDCK_(NBL-2, RRID:CVCL_0422)Experimental Models: Organisms/Strains Sentences Resources SARS-CoV-2 B.1.617.2 (delta) variant was acquired from the California Department of Public Health, cultured in Vero-TMPRSS2 cells. SARS-CoV-2 B.1.617.2suggested: NoneSoftware and Algorithms Sentences Resources Confocal imaging was performed using a Nikon A1R laser scanning confocal microscope with NIS-Elements software and a 16X LWD water dipping objective. NIS-Elementssuggested: None50 – 100 µm-thick images with a z-step of 1.5 µm were taken and analyzed using Imaris (Bitplane). Imarissuggested: NoneData were collected using the BD LSRII Cytometer and analyzed using FlowJo version 10 (BD Biosciences). FlowJosuggested: (FlowJo, RRID:SCR_008520)Briefly, raw gene-expression fastqs were aligned to the GRCh38 reference genome annotated with Ensembl v85, and Lipid Hashtag fastqs were processed to count the incidences of each expected index per cell. Ensemblsuggested: NoneData quality control and normalization: The gene expression count matrices were normalized, and variance stabilized using negative binomial regression using the scTransform algorithm21 in the Seurat package. Seuratsuggested: (SEURAT, RRID:SCR_007322)Statistical analyses: Statistical analysis was performed using GraphPad Prism v7.0e. 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.
Results from scite Reference Check: We found no unreliable references.
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