title: Genome-wide fitness profiling reveals adaptations required by Haemophilus in coinfection with influenza A virus in the murine lung.
abstract: Bacterial coinfection represents a major cause of morbidity and mortality in epidemics of influenza A virus (IAV). The bacterium Haemophilus influenzae typically colonizes the human upper respiratory tract without causing disease, and yet in individuals infected with IAV, it can cause debilitating or lethal secondary pneumonia. Studies in murine models have detected immune components involved in susceptibility and pathology, and yet few studies have examined bacterial factors contributing to coinfection. We conducted genome-wide profiling of the H. influenzae genes that promote its fitness in a murine model of coinfection with IAV. Application of direct, high-throughput sequencing of transposon insertion sites revealed fitness phenotypes of a bank of H. influenzae mutants in viral coinfection in comparison with bacterial infection alone. One set of virulence genes was required in nonvirally infected mice but not in coinfection, consistent with a defect in anti-bacterial defenses during coinfection. Nevertheless, a core set of genes required in both in vivo conditions indicated that many bacterial countermeasures against host defenses remain critical for coinfection. The results also revealed a subset of genes required in coinfection but not in bacterial infection alone, including the iron-sulfur cluster regulator gene, iscR, which was required for oxidative stress resistance. Overexpression of the antioxidant protein Dps in the iscR mutant restored oxidative stress resistance and ability to colonize in coinfection. The results identify bacterial stress and metabolic adaptations required in an IAV coinfection model, revealing potential targets for treatment or prevention of secondary bacterial pneumonia after viral infection.
authors.affiliation: Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605.
authors.forename: Sandy M
authors.initials: SM
authors.lastname: Wong
authors.forename: Mariana
authors.initials: M
authors.lastname: Bernui
authors.forename: Hao
authors.initials: H
authors.lastname: Shen
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Akerley
mesh_terms: D000220:Adaptation, Biological / Q000235:genetics*; D000818:Animals; D060085:Coinfection / Q000382:microbiology* / Q000821:virology; D004251:DNA Transposable Elements / Q000235:genetics; D056084:Genetic Fitness / Q000235:genetics*; D006190:Haemophilus / Q000235:genetics*; D006192:Haemophilus Infections / Q000235:genetics / Q000503:physiopathology*; D059014:High-Throughput Nucleotide Sequencing; D009980:Influenza A virus; D008168:Lung / Q000382:microbiology / Q000821:virology; D051379:Mice; D009976:Orthomyxoviridae Infections / Q000382:microbiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D014157:Transcription Factors
keywords: functional genomics; transposon-insertion sequencing