Journal: ImmunoHorizons

Article Title: IL-13 protects epithelial cells from SARS-CoV-2 infection by inhibiting ACE2-mediated virus binding and cell entry

doi: 10.1093/immhor/vlaf056

Figure Lengend Snippet: IL-13 inhibits ACE2 transcription in epithelial cells. ACE2 mRNA levels were measured by RT-qPCR in Vero E6 epithelial cells treated with IL-13 (10 ng/ml) or PBS for 24, 48, or 72 h (A), or in Calu3 cells treated with IL-13 (10 ng/ml) or PBS for 24 h (B), or in human primary nasal (C) and bronchial (D) epithelial cells treated with IL-13 (10 ng/ml) or PBS for 24 h. The mRNA levels at each time point were normalized by GAPDH (A, B) or PPIA (C, D) and are shown relative to PBS. Shown are means ± SEM from 1 representative experiment out of 2. Each data point represents a replicate culture [ n = 5–6/condition (A), n = 3/condition (B)]. In (C, D) each data point represents a single donor ( n = 8–12) each tested in triplicate. An unpaired 2-tailed t test was used for statistical analysis. An unpaired 2-tailed t test was used for statistical analysis.

Article Snippet: HEK293T/ACE2 cells stably transduced with human ACE2 were purchased from GeneCopoeia (SL221, Rockville, Maryland, USA).

Techniques: Quantitative RT-PCR

Journal: ImmunoHorizons

Article Title: IL-13 protects epithelial cells from SARS-CoV-2 infection by inhibiting ACE2-mediated virus binding and cell entry

doi: 10.1093/immhor/vlaf056

Figure Lengend Snippet: IL-13 downregulates surface ACE2 expression in epithelial cells. Vero (A, B) or Calu-3 (C) cells were treated with IL-13 (2 or 10 ng/ml) for 72 h and assessed for ACE2 protein expression by flow cytometry using ACE2-specific monoclonal antibodies (clone 36 from Sino Biological in A and B, clone 535919 from R&D systems in C) and relevant isotype controls. Shown are a representative flow cytometry histogram for Vero cells (A) (shaded area: isotype control) and proportions of ACE2 + cells in IL-13-treated cells relative to PBS (B, C). The data are from 1 representative experiment out of 3 and show means ± SEM. Data points represent replicate cultures ( n = 4/condition). An unpaired 2-tailed t test was used for statistical analysis.

Article Snippet: HEK293T/ACE2 cells stably transduced with human ACE2 were purchased from GeneCopoeia (SL221, Rockville, Maryland, USA).

Techniques: Expressing, Flow Cytometry, Bioprocessing, Control

Journal: ImmunoHorizons

Article Title: IL-13 protects epithelial cells from SARS-CoV-2 infection by inhibiting ACE2-mediated virus binding and cell entry

doi: 10.1093/immhor/vlaf056

Figure Lengend Snippet: IL-13 does not affect ACE2 expression and S protein-pseudotyped lentiviral entry into HEK293T/ACE2 cell transfectants. HEK293T/ACE2 cells were treated with 10 ng/ml IL-13 for 72 h and surface ACE2 expression was evaluated by flow cytometry with ACE2-specific monoclonal antibodies. Shown are representative flow cytometry histograms (A), proportions (B) and MFI (C) of ACE2+ cells in cultures treated with IL-13 or PBS. Data are from 1 experiment ( n = 5–7 replicates per condition). An unpaired, two-tailed t test was used for statistical analysis. ACE2 -transfected HEK293T/ACE2 cells were treated with IL-13 (10 ng/ml) (E, H) or PBS (D, G) for 48 h, washed and transduced with lentiviral particles pseudotyped with SARS-CoV-2 S protein (0.2 × 10 4 TU: D, E) or VSV-G protein (2 × 10 4 TU: G, H). Shown are frames representative of GFP fluorescence under the indicated experimental conditions. GFP fluorescence was assessed under a microscope (10× magnification) 72 h post transduction and quantified using Slide Analyzer (F, I). The scale bars represent 200 µm. Results represent the mean ± SEM of 20 (SARS-CoV-2-S-pseudotyped virus) or 10 (VSV-G-pseudotyped virus) random frames from 1 out of 2 representative experiments. A Wilcoxon 2-sample test was used for statistical analysis.

Article Snippet: HEK293T/ACE2 cells stably transduced with human ACE2 were purchased from GeneCopoeia (SL221, Rockville, Maryland, USA).

Techniques: Expressing, Flow Cytometry, Bioprocessing, Two Tailed Test, Transfection, Transduction, Fluorescence, Microscopy, Virus