Fig. 4: Short ACE2 is upregulated in response to IFN and RV16 infection but not SARS-CoV-2 infection.

a–f, Box (median and quartiles) and whisker (5th to 95th percentiles) plots plus individual data points of relative expression (\(2^{-\Delta \Delta {\rm{C}}_{{\rm{t}}}}\)) of transcripts/genes measured by RT–qPCR. a, Relative expression of total (left), long (middle) and short (right) ACE2 transcripts in undifferentiated PBECs treated with IFN-α (n = 3 donors), IFN-β (n = 7 donors), IFN-γ (n = 4 donors) or IFN-λ (n = 4 donors) for 24 h at indicated doses. Data were analyzed using a two-tailed, Mann–Whitney U-test, compared with control. Total ACE2: IFN-α 1,000: P = 0.0083; IFN-β 100: P = 0.0006; IFN-β 1,000: P = 0.0006; IFN-γ 100: P = 0.030; IFN-γ 1,000: P = 0.0030; IFN-λ 100: P = 0.0030. Long ACE2: IFN-α 100: P = 0.0083; IFN-α 1,000: P = 0.0083; IFN-γ 1,000: P = 0.0030; IFN-λ 100: P = 0.0030. Short ACE2: IFN-β 100: P = 0.0006; IFN-γ 1,000: P = 0.030; IFN-λ 100: P = 0.0030. b,c, Relative expression of total, long and short ACE2 in in vitro differentiated NECs (n = 11 individual donors) (b) and PBECs (n = 13 individual donors) (c) 24 h after infection with RV16 or UV-RV16. Data were analyzed using a two-tailed Wilcoxon’s test, compared with UV-RV16. NECS: total ACE2: P = 0.0010; long ACE2: P = 0.0010; short ACE2: P = 0.0010. PBECs: total ACE2: P = 0.0010; long ACE2: P = 0.2510; short ACE2: P = 0.0007. d–f, Normalized expression of CoV-N1 (d), relative expression of ACE2 transcripts (e) and MX1 and IP10 (f) in differentiated BCi-NS1.1 cells 1 and 72 h after infection with SARS-CoV-2 strain nCoV/Victoria/1/2020 (n = 4 independent experiments). CoV-N1, MX1 and IP10 expression data were analyzed using paired, two-tailed Student’s t-test (CoV-N1, P = 0.0852). ACE2 data were analyzed using nonparametric, two-tailed, Wilcoxon’s test. g, Dot plot showing ratio of short to long ACE2 expression (Δshort–long −ΔC_t) in in vitro differentiated PBECs from healthy (n = 13 donors) or severely asthmatic (n = 12 donors) donors 24 h after infection with RV16 or UV-RV16. The two-tailed Wilcoxon’s test was used for paired and the two-tailed Mann–Whitney U-test for unpaired samples. UV-RV16 versus RV16 in healthy controls: P = 0.0005; in severe asthma: P = 0.0020. UV-RV16 in healthy controls versus severe asthma: P = 0.0138; RV16 in healthy controls versus severe asthma: P = 0.0114. h, Dot plot showing correlation of relative expression of IFNB1 and short ACE2 (left) or long ACE2 (right) in response to RV16 infection. Data were analyzed using two-tailed Spearman’s rank correlation coefficient: short ACE2: r = 0.5765; P = 0.0032; long ACE2: r = 0.05391; P = 0.8024.