Figure 1.
Intravital of the lungs in transgenic mice infected with SARS-CoV-2. Transgenic mice expressing human ACE2 (hACE2) driven by the keratin 18 (hACE2 K18) or CAG (hACE2 AC70) promoter were infected with NG SARS-CoV2 via different doses and routes. (A) Percentage of body weight change in hACE2 AC70 and hACE2 K18 mice infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN. (B) Percentage of body weight change in hACE2 AC70 mice infected with 2.5 × 103 or 2.5 × 102 pfu NG-SARS-CoV-2 via IN. (C) Percentage of body weight change in hACE2 AC70 mice infected with 2.5 × 104 pfu NG-SARS-CoV-2 via oropharyngeal, intratracheal, or intraperitoneal administration. (A-C) Each line represents 1 mouse. (D) Intravital microscopy (IVM) images (original magnification ×20) of the lungs of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN showing neutrophils (anti-Ly6G, red) in the vasculature (day 6 after infection). Scale bar, 12 μm. The graph represents the quantification of neutrophils per FOV in the lung after IVM. (E) Quantification of the number of neutrophils adhered in the capillaries of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN after IVM (day 6 after infection). (F) Quantification of neutrophils in the lungs of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN after flow cytometry (day 5 after infection). (G) Hematoxylin and eosin staining of lung sections from hACE2 AC70 mice infected with NG-SARS-CoV-2. (H) Quantification of neutrophils in the BAL of hACE2 AC70 mice uninfected, infected with 2.5 × 104 pfu NG-SARS-CoV-2 or influenza (500 pfu), or treated with the neutrophil chemokine KC via IN, after flow cytometry (day 5 after tinfection). (I) Quantification of inflammatory monocytes in the BAL of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 or influenza (500 pfu) via IN, after flow cytometry (day 5 after infection). Data (as applicable) are represented as mean ± SD. Unpaired Student t test was performed for graphs with 2 groups, and one-way analysis of variance test was performed for multiple comparisons in graphs with >2 groups. ∗∗P < .01; ∗∗∗P < .001. IN, intranasal administration; NG, neon-green; ns, not significant; SD, standard deviation.

Intravital of the lungs in transgenic mice infected with SARS-CoV-2. Transgenic mice expressing human ACE2 (hACE2) driven by the keratin 18 (hACE2 K18) or CAG (hACE2 AC70) promoter were infected with NG SARS-CoV2 via different doses and routes. (A) Percentage of body weight change in hACE2 AC70 and hACE2 K18 mice infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN. (B) Percentage of body weight change in hACE2 AC70 mice infected with 2.5 × 103 or 2.5 × 102 pfu NG-SARS-CoV-2 via IN. (C) Percentage of body weight change in hACE2 AC70 mice infected with 2.5 × 104 pfu NG-SARS-CoV-2 via oropharyngeal, intratracheal, or intraperitoneal administration. (A-C) Each line represents 1 mouse. (D) Intravital microscopy (IVM) images (original magnification ×20) of the lungs of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN showing neutrophils (anti-Ly6G, red) in the vasculature (day 6 after infection). Scale bar, 12 μm. The graph represents the quantification of neutrophils per FOV in the lung after IVM. (E) Quantification of the number of neutrophils adhered in the capillaries of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN after IVM (day 6 after infection). (F) Quantification of neutrophils in the lungs of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 via IN after flow cytometry (day 5 after infection). (G) Hematoxylin and eosin staining of lung sections from hACE2 AC70 mice infected with NG-SARS-CoV-2. (H) Quantification of neutrophils in the BAL of hACE2 AC70 mice uninfected, infected with 2.5 × 104 pfu NG-SARS-CoV-2 or influenza (500 pfu), or treated with the neutrophil chemokine KC via IN, after flow cytometry (day 5 after tinfection). (I) Quantification of inflammatory monocytes in the BAL of hACE2 AC70 mice uninfected or infected with 2.5 × 104 pfu NG-SARS-CoV-2 or influenza (500 pfu) via IN, after flow cytometry (day 5 after infection). Data (as applicable) are represented as mean ± SD. Unpaired Student t test was performed for graphs with 2 groups, and one-way analysis of variance test was performed for multiple comparisons in graphs with >2 groups. ∗∗P < .01; ∗∗∗P < .001. IN, intranasal administration; NG, neon-green; ns, not significant; SD, standard deviation.

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