Fig. 2: Establishing severe influenza on chip.
From: An immune-competent lung-on-a-chip for modelling the human severe influenza infection response
a, Representative RNA-FISH images of the epithelial layer of the device, showing nuclei in blue and viral H1N1 RNA in yellow for the LOC and IC-LOC at MOI = 0, 0.1, 1 and 10. b, Quantification of the viral RNA counts per cell, as determined via RNA-FISH. N = 3 independent devices. Statistical significance determined via one-way ANOVA with Kruskal–Wallis test, and P values are shown. c, Relative quantification of infectious viral titre via a ViralTox Glo assay. Relative luminescence (RLU) corresponds to ATP activity of H1N1-susceptible MDCK cells cultured with the cell lysis of uninfected devices or LOC, Mϕ-LOC or IC-LOC devices infected at MOI = 10. N = 10 independent devices. d, Quantification of antigen (HA) concentration in the cell lysis of LOC, Mϕ-LOC or IC-LOC devices infected at MOI = 10. N = 6 independent devices. e, Quantification of epithelial damage via LDH activity and airway permeability in the LOC in response to H1N1 infection at MOI. N = 8 independent devices. f, Quantification of epithelial damage via LDH activity and airway permeability in the IC-LOC in response to H1N1 infection at varying MOIs. N = 3 independent devices. g, LEGENDplex analysis of the cytokine release profiles in the LOC and IC-LOC 24 hpi at varying MOIs. Heat map shows the fold change from time zero (pre-infection). N = 4 independent devices. h,i, Simple linear regression correlation of airway and vascular levels of CXCL10 (h) and IFN-λ1 (i) in the IC-LOC. N = 14 independent devices. j,k, Comparison of cytokine release profiles in the airway layer of the IC-LOC (j) and LOC (k) to that of BAL fluid from patients with moderate and severe influenza. Fold change from uninfected/healthy is shown. Purple bands denote the range of patient data with severe influenza, while teal bands denote the range of patients with moderate influenza. N = 3 independent devices. All immune-competent devices use cells from the same immune cell donor. All statistical significance determined via one-way ANOVA followed by Tukey’s multiple comparisons test unless otherwise noted. Data are presented as mean values ± s.d.
