Figure 5.
Effect of prior RV infection on ISG induction and SARS-CoV-2 replication in human airway epithelial organoids.(A) Timing of infection of epithelial organoids with RV followed by SARS-CoV-2. (B) Expression of ISGs in airway epithelial organoids 3 d after RV infection, relative to mRNA for the housekeeping gene HPRT. (C) SARS-CoV-2 viral RNA at 24, 48, and 72 h after infection, with or without RV preinfection. P value represents difference between exponential growth curves fit to the data. (D) ELISA for IFNλ1 in basolateral media from uninfected organoid cultures or cultures infected with SARS-CoV-2 with or without RV preinfection. (E) ELISA for IFNβ in basolateral media from uninfected organoid cultures or cultures infected with SARS-CoV-2 with or without RV preinfection. (F) Expression of ISGs at 24, 48, and 72 h after SARS-CoV-2 infection, with or without RV preinfection, expressed as FC from uninfected cells. (G) Single-cell sequencing of human airway epithelial cell organoids, mock or 5 d after RV infection. Red and orange dots indicate 70/4,200 cells with detectable viral RNA at this time point in RV-infected cultures. tSNE plots show expression of mRNA for ISGs in mock and infected cultures at the same time point. Both conditions are from a single experiment using organoids from the same donor, with data from 8,711 cells (mock, n = 4,200 cells; infected, n = 4,511 cells). PNEC, pulmonary neuroendocrine cell. For B, bars show mean and SEM of four replicates per condition. Data are representative of three independent experiments with organoids from different primary cell donors. For C–F, bars show mean and SEM of four to six biological replicates per condition. Data are representative of two independent experiments with organoids from different primary cell donors. For B–F, significant differences between conditions were evaluated by the Mann–Whitney test (*, P = 0.0317; #, P = 0.0286; ##, P = 0.0159; **, P = 0.0079).

Effect of prior RV infection on ISG induction and SARS-CoV-2 replication in human airway epithelial organoids. (A) Timing of infection of epithelial organoids with RV followed by SARS-CoV-2. (B) Expression of ISGs in airway epithelial organoids 3 d after RV infection, relative to mRNA for the housekeeping gene HPRT. (C) SARS-CoV-2 viral RNA at 24, 48, and 72 h after infection, with or without RV preinfection. P value represents difference between exponential growth curves fit to the data. (D) ELISA for IFNλ1 in basolateral media from uninfected organoid cultures or cultures infected with SARS-CoV-2 with or without RV preinfection. (E) ELISA for IFNβ in basolateral media from uninfected organoid cultures or cultures infected with SARS-CoV-2 with or without RV preinfection. (F) Expression of ISGs at 24, 48, and 72 h after SARS-CoV-2 infection, with or without RV preinfection, expressed as FC from uninfected cells. (G) Single-cell sequencing of human airway epithelial cell organoids, mock or 5 d after RV infection. Red and orange dots indicate 70/4,200 cells with detectable viral RNA at this time point in RV-infected cultures. tSNE plots show expression of mRNA for ISGs in mock and infected cultures at the same time point. Both conditions are from a single experiment using organoids from the same donor, with data from 8,711 cells (mock, n = 4,200 cells; infected, n = 4,511 cells). PNEC, pulmonary neuroendocrine cell. For B, bars show mean and SEM of four replicates per condition. Data are representative of three independent experiments with organoids from different primary cell donors. For C–F, bars show mean and SEM of four to six biological replicates per condition. Data are representative of two independent experiments with organoids from different primary cell donors. For B–F, significant differences between conditions were evaluated by the Mann–Whitney test (*, P = 0.0317; #, P = 0.0286; ##, P = 0.0159; **, P = 0.0079).

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