a549 human type i alveolar cell line  (ATCC)

Journal: Cells

Article Title: Eosinophil Responses at the Airway Epithelial Barrier during the Early Phase of Influenza a Virus Infection in C57BL/6 Mice

doi: 10.3390/cells10030509

Figure Lengend Snippet: Eosinophils protect the epithelial cell barrier during influenza virus infection. ( A ) Representative images of A549 cell monolayers in each culture condition. ( B ) Percentage of dead A549 cells in the culture conditions. ( C ) Influenza internal protein (PB1) expressing A549 cells and ( D ) bone marrow-derived eosinophils in each culture condition. ( E ) Influenza A virus (pH1N1) titer in the supernatants. Experiments independently repeated three times for rigor and reproducibility. Data are represented as the mean and standard deviation of n = 5–6 samples analyzed by two-way ANOVA with Sidak’s multiple comparisons test. Differences are significant ( p < 0.05) when letters above bars are dissimilar. ** p < 0.01 and *** p < 0.001. BMdEos—bone marrow-derived eosinophils; d/D—direct contact; id/ID—indirect contact.

Article Snippet: Owing to expression of both α-2,6 and α-2,3 linked sialic acid residues, A549 human type I alveolar cell line derived from a carcinoma patient (ATCC) is suitable and has been used to study the pathogenesis of influenza viruses for years [ , ].

Techniques: Virus, Infection, Expressing, Derivative Assay, Standard Deviation

Journal: Cells

Article Title: Eosinophil Responses at the Airway Epithelial Barrier during the Early Phase of Influenza a Virus Infection in C57BL/6 Mice

doi: 10.3390/cells10030509

Figure Lengend Snippet: Eosinophils affect the epithelial transcriptome during influenza virus infection. ( A ) Microarray analysis of the A549 cell transcriptome during mock or influenza virus (IAV) infection when in direct (d) or indirect (id) contact with bone marrow derived eosinophils (BMdEos). Heat map contains log 2 signal values for 1969 individual samples normalized to the row mean log 2 signal value for samples M1-M4 (Mock-infected, no BMdEos). Dendrogram (left side) shows hierarchical clustering of genes by complete linkage based on euclidean distance. ( B ) The number of differentially expressed genes (DEGs) in each group relative to the average expression in the M group shown as a stacked bar representing the mean and standard deviation. Up- and down-regulated genes were compared across the groups by 2-way ANOVA with Tukey’s multiple comparisons test where letters above bars represent p < 0.05 when different. Protein interaction networks generated by STRINGdb are shown for: ( C ) Top 25% of upregulated genes (281 DEGs) between mock-infected and IAV-infected epithelial cells with no BMdEos exposure; network required an edge confidence score ≥0.700 and contains an additional 20 s shell nodes (white). ( D ) Top 25% of downregulated genes (212 DEGs) between mock-infected and IAV-infected epithelial cells with no BMdEos exposure; network required an edge confidence score ≥0.400 and contains an additional 30 s shell nodes (white). ( E ) Top 50% of downregulated genes (267 DEGs) between mock-infected and IAV-infected epithelial cells with direct BMdEos exposure; network required an edge confidence score ≥0.400 and contains an additional 55 s shell nodes (white). ( F ) All upregulated genes (91 DEGs) between mock-infected and IAV-infected epithelial cells with direct BMdEos exposure; network required an edge confidence score ≥0.400 and contains an additional 65 s shell nodes (white). For closer inspection, individual PNG files of the protein interaction networks shown in C-F are provided in as . Please note that gene expression data and pathway analysis results of these data are provided in . M—mock, V—virus, d/D—direct culture, id/ID—indirect culture.

Article Snippet: Owing to expression of both α-2,6 and α-2,3 linked sialic acid residues, A549 human type I alveolar cell line derived from a carcinoma patient (ATCC) is suitable and has been used to study the pathogenesis of influenza viruses for years [ , ].

Techniques: Virus, Infection, Microarray, Derivative Assay, Expressing, Standard Deviation, Generated, Gene Expression

Journal: Cells

Article Title: Eosinophil Responses at the Airway Epithelial Barrier during the Early Phase of Influenza a Virus Infection in C57BL/6 Mice

doi: 10.3390/cells10030509

Figure Lengend Snippet: Cross activation of epithelial and eosinophils occurs in response to influenza A virus. ( A ) A549 epithelial cells and bone marrow-derived eosinophils were cultured directly/indirectly after infecting epithelial cells with pH1N1 influenza A virus and each cell type was analyzed by flow cytometry. Illustration with BioRender. ( B ) Surface expression of markers on epithelial cells and eosinophils. Data represented as the mean and standard deviation of n = 5–6 samples analyzed by two-way ANOVA with Tukey’s multiple comparisons test of one of three independent studies. Differences are significant ( p < 0.05) when letters above bars are dissimilar. BMdEos—bone marrow-derived eosinophils; d—direct contact; id—indirect contact.

Article Snippet: Owing to expression of both α-2,6 and α-2,3 linked sialic acid residues, A549 human type I alveolar cell line derived from a carcinoma patient (ATCC) is suitable and has been used to study the pathogenesis of influenza viruses for years [ , ].

Techniques: Activation Assay, Virus, Derivative Assay, Cell Culture, Flow Cytometry, Expressing, Standard Deviation