Dissection of key factors correlating with H5N1 avian influenza virus driven inflammatory lung injury of chicken identified by single-cell analysis
Fig 5
The key immune cells and genes contribute to the H5N1 AIV-driven pneumonia.
(A) The normalized expression (UMI counts) of pro-inflammatory genes including IL6, CCL19, CCL4, CXCL13, IL1β, IL8, and TNFAIP3 in different cell populations. (B) Pearson’s Correlation analysis between the SMART-Seq2 data of macrophages (KUL01+CLASS II+) and lung clusters in scRNA-seq based on levels of gene expression. (C) Analysis expression of 21 differentially expressed genes (DEGs) in macrophages of single-cell lung suspensions from H9N2 AIV infected chickens at 3DPI by qRT-PCR. (D) Analysis expression of 21 DEGs in macrophages of single-cell lung suspensions from H5N1 AIV infected chickens at 1DPI by qRT-PCR. Total RNA of sorted macrophages was extracted from three chickens of the two infection groups and control groups, respectively. The data were collected from three biological samples, and each sample was tested in triplicate. The results are presented as mean ± SEM. Statistical comparisons were performed with paired t-test, and significance was assessed as P-values using GraphPad Prism. *P < 0.05, **P < 0.01, ***P < 0.001. (E) Predicted interaction map through CCL4, CCL19, and CXCL13 after H5N1 AIV challenge, respectively. Purple line indicates interactions emerging only after H5N1 AIV challenge.