Host-pathogen interaction profiling of nontypeable Haemophilus influenzae and Moraxella catarrhalis coinfection of bronchial epithelial cells.
Chronic obstructive pulmonary disease (COPD) is a progressive inflammatory lung disease and the third leading cause of death globally. Nontypeable Haemophilus influenzae (NTHi) and Moraxella catarrhalis (Mcat) are common pathogens in individuals with COPD. Acquisition of NTHi or Mcat can cause acute exacerbations of COPD. NTHi and/or Mcat can also persist for months in the lower airways and lead to chronic inflammation. We hypothesized that infection by NTHi or Mcat, singly or during coinfection, requires regulation of specific bacterial and host cell pathways. We investigated this phenomenon using an in vitro cell culture model consisting of lung carcinoma H292 cell lines, infected with NTHi, Mcat, or both species. Samples were fractionated into “apical fluid”, containing free floating bacteria, and adhered/invaded bacteria on or within H292 cells. We used transcriptomic profiling with RNA-seq and various bioinformatic analyses to disentangle host-pathogen interactions in epithelial cell infection from the perspective of each species. Several biological pathways were differentially regulated across all conditions (31, NTHi; 22, Mcat; and 169, Human). NTHi transcriptomic profiles differed during single and coinfection; examples included downregulation of iron-sulfur metabolism (IscR regulon) and differential regulation of quorum sensing in coinfection compared to single infection.  Mcat was comparatively less affected by the presence of NTHi during coinfection.  H292 epithelial cells responded broadly to all infections with distinct responses to single and coinfection. Enriched host pathways included influenza/interferon/Wnt and proinflammatory responses. These findings suggest common and distinct processes involved in NTHi and/or Mcat induced COPD pathogenesis and have implications for therapeutic intervention. Overall design: To investigate the host-pathogen interactions of COPD pathogens, nontypable Haemophilus influenzae (NTHi) and Moraxella catarrhalis, with human epithelial cells NCI-H292. We generated single infections and coinfections of H292 cells, fractioned samples into apical fluid or adherent/invaded cells, as well as bacterial controls. We then performed comparative gene expression profiling analysis using multi-species RNA-seq.