Transcriptome Profiles of Human Lung Epithelial Cells A549 Interacting with Aspergillus fumigatus by RNA-Seq

• Published in: PloS one 2015-08, Vol.10 (8), p.e0135720-e0135720
• Main Authors: Chen, Fangyan, Zhang, Changjian, Jia, Xiaodong, Wang, Shuo, Wang, Jing, Chen, Yong, Zhao, Jingya, Tian, Shuguang, Han, Xuelin, Han, Li
• Format: Article
• Language: English
• Subjects: Alveolar Epithelial Cells - metabolism; Alveoli; Aspergillus; Aspergillus fumigatus; Aspergillus fumigatus - physiology; Cell Line; Chemotaxis; Comparative analysis; Conidia; Disease control; Disease prevention; EGR-1 protein; EGR-4 protein; Enrichment; Epithelial cells; Gene expression; Gene Expression Profiling; Gene Expression Regulation; Genes; Genomes; Gram-positive bacteria; Hemopoiesis; High-Throughput Nucleotide Sequencing; Hospitals; Human behavior; Humans; Immune response; Immune system; Immunology; Infections; Inflammation; Inflammatory response; Interleukin 6; Interleukin 8; Internalization; Ion transport; Kinases; Lung - metabolism; Lungs; MAP kinase; MAP Kinase Signaling System; Phosphatase; Proteins; Respiratory Mucosa - metabolism; Ribonucleic acid; RNA; Sepsis; Spores, Fungal; Studies; Technology assessment; Transcription; Transcriptome; Tumor necrosis factor-α
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0135720

Lung epithelial cells constitute the first defense line of host against the inhaled Aspergillus fumigatus; however, the transcriptional response of human alveolar type II epithelial cells was still unclear. Here we used RNA-Seq technology to assess the transcriptome profiles of A549 cells following direct interaction with conidia of A. fumigatus. The total number of identified genes was 19118. Compared with uninfected A549 cells, 459 genes were differentially expressed in cells co-incubated with conidia for 8 h, including 302 up-regulated genes and 157 down-regulated genes. GO and KEGG pathway enrichment analysis showed that most of the up-regulated genes were related to immune response, chemotaxis and inflammatory response and enriched in cytokine-cytokine receptor interaction, JAK-STAT and MAPK signaling pathways. The down-regulated genes were mainly enriched for terms associated with development, hemopoiesis and ion transport. Among them, EGR4 and HIST1H4J gene had the maximum of fold change in up-regulated and down-regulated genes, respectively. Fourteen up-regulated genes and three down-regulated genes were further validated and significant increase on expression of IL-6, IL-8 and TNF-α in A549 cells were confirmed by qRT-PCR during the interaction of A549 cells with A. fumigatus. Besides, western blot showed that expression of two proteins (ARC, EGR1) significantly increased in A549 cells during interaction with A. fumigatus conidia for 8h. Interference of endogenous expression of ARC or EGR1 protein in A549 cells reduced the internalization of A. fumigatus. These results provided important insights into dynamic changes of gene expression in lung epithelial cells, especially its strong immunological response against A. fumigatus infection.