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The initial stages of the interaction between the host and  Aspergillus fumigatus  at the alveolar surface of the human lung are critical in the establishment of aspergillosis. Using an  in vitro  bilayer model of the alveolus, including both the epithelium (human lung adenocarcinoma epithelial cell line, A549) and endothelium (human pulmonary artery epithelial cells, HPAEC) on transwell membranes, it was possible to closely replicate the  in vivo  conditions. Two distinct sub-groups of dendritic cells (DC), monocyte-derived DC (moDC) and myeloid DC (mDC), were included in the model to examine immune responses to fungal infection at the alveolar surface. RNA in high quantity and quality was extracted from the cell layers on the transwell membrane to allow gene expression analysis using tailored custom-made microarrays, containing probes for 117 immune-relevant genes. This microarray data indicated minimal induction of immune gene expression in A549 alveolar epithelial cells in response to germ tubes of  A. fumigatus . In contrast, the addition of DC to the system greatly increased the number of differentially expressed immune genes. moDC exhibited increased expression of genes including  CLEC7A, CD209  and  CCL18  in the absence of  A. fumigatus  compared to mDC. In the presence of  A. fumigatus , both DC subgroups exhibited up-regulation of genes identified in previous studies as being associated with the exposure of DC to  A. fumigatus  and exhibiting chemotactic properties for neutrophils, including  CXCL2, CXCL5, CCL20 , and  IL1B . This model closely approximated the human alveolus allowing for an analysis of the host pathogen interface that complements existing animal models of IA.

plos one

Gene Expression Profiles of Human Dendritic Cells Interacting with Aspergillus fumigatus in a Bilayer Model of the Alveolar Epithelium/Endothelium Interface