Extended Data Fig. 8: ALOX15-deficient mice are more susceptible to a low dose of IAV.
From: Neonatal imprinting of alveolar macrophages via neutrophil-derived 12-HETE
(a-c) WT and Alox15−/− animals were infected with IAV (50 pfu, in). (a) At various days post-infection, expression of ALOX15 was evaluated by flow cytometry (ICS) in CD45− cells as well as various leukocyte populations. The expression was normalized to Alox15−/− mice used as controls for gating (n = 4/5/3/5/group). (b) Levels of 12-HETE (left, n = 5/10/8/13/group) and 15-HETE (right, n = 6/12/8/13/group) in the BAL at various days post-infection. (c-i) WT and Alox15−/− animals were infected with IAV (20 pfu, in). (c) Pulmonary viral loads at day 3 post-infection (n = 4/group). (d-h) BAL levels of IFNβ (d) (0: n = 3/group, 3: n = 5/group) and PGE2 (e) (n = 5/group) at day 3 post-infection. (f) BAL levels of CCL2 (left) and CXCL1/KC (right) at various days post-infection (WT: n = 3/5/9/5/6/3/group, Alox15−/−: n = 3/5/8/6/5/3 mice/group). (g) BAL numbers of AM (left), monocytes (middle) and neutrophils (right) at various days post-infection (WT: n = 3/5/5/5/6/3/timepoint, Alox15−/−: n = 3/5/5/6/5/3/timepoint). (h) Total BAL protein levels at various days post-infection (WT: n = 3/5/9/5/6/3/group, Alox15−/−: n = 3/5/7/6/5/3/group). (i) Representative micrographs of H&E lung sections at various days post-infection. (j) Alox15−/− mice were treated daily with exogenous 12-HETE or 15-HETE or vehicle. Lungs were harvested at day 3 post-infection for assessment of pulmonary viral loads (n = 8/8/7/8/group). (k) Alox15−/− mice were treated daily with exogenous 12-HETE or 15-HETE or vehicle starting on the day of infection (IAV, 90 pfu, in) for 3 days. The survival was monitored overtime (n = 8/10/9/9/group). (l) Weight loss curves after SARS-CoV-2 infection (k18-hACE2 Alox15+/+: n = 19, k18-hACE2 Alox15−/−: n = 17). Data are presented as mean ± s.e.m and are from one (a, c-e, k) or pooled from two (b, f-h, j, l) independent experiments or representative of 5 (i) biological replicates. Data were analyzed using two-tailed unpaired t-test (c, e), one-way ANOVA followed by Tukey’s multiple comparisons test (b, j), two-way ANOVA followed by Sidak’s multiple comparisons test (d, f-h) or log-rank test (k).
