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[Preprint]. 2024 Mar 27:2024.03.27.586885. [Version 1] doi: 10.1101/2024.03.27.586885

Figure 5: Recent IFN-I dependent and - independent inflammatory conditioning of the lung promotes SCV2 replication control at the tissue level.

Figure 5:

(A) Heatmap of fold change in the geometric mean fluorescence intensity (gMFI) of IFN-inducible surface marker (ISM) expression of Sca-1 and CD317 measured by flow cytometry on lung epithelial cell (EC) subsets from lungs of mice treated with various inflammatory or infectious stimuli compared to those from PBS control animals at the indicated time points without SCV2 infection, n=5-14, data is pooled from 2 – 4 independent experiments, for all conditions except OVA/Alum which was done once. (B) Ifnar1−/−, (C) Tnfrsf1a−/−, or (D) Il1r1−/− mice were i.ph. treated with PBS, 10μg CpG or 50μg Pm3 one week before SCV2 (B.1.351) infection. Viral loads in lungs were quantified by TCID50 or sub-gRNA SCV2 E qPCR, n=10-26, data are combined from 2 – 3 independent experiments each (E) Schematic of K18-hACE2 Tg mice administered 5μg recombinant mouse TNFα (rmTNFα) or 2.0x104U recombinant mouse IFNβ (rmIFNβ) once, one week before infection with SCV2 (USA-WA1/2020) and lung viral titers, n=7-10, two independent experiments (F) Fold change in gMFI of Sca-1 and CD317 measured by flow cytometry on lung EC subsets of mice treated one week prior with rmTNFα or rmIFNβ, data is pooled from three independent experiments, n= 15-22 , *= p<0.05, **=p<0.01, ***=p<0.001, if not indicated, differences were not significant. (G) WT or Il1r1−/− mice given 200U recombinant mouse IL-1α (rmIL-1α) or IL-1β (rmIL-1β) i.ph. one week before SCV2 (B.1.351) infection and ung viral titers, n=8-10, data are combined from two independent experiments. (H) Fold change in gMFI of Sca-1 and CD317 measured by flow cytometry on EC subsets from lungs of mice treated one week prior with 200U rmIL-1α + rmIL-1β (100U each), n= 9-10, data is pooled from two independent experiments. Geometric mean, statistical significance determined by two-tailed Mann-Whitney test, LD= limit of detection, *= p<0.05, **=p<0.01, ***=p<0.001, n.s.= not significant, if not indicated differences were not significant, white arrows indicate direction of fold change.