Transposon Mutagenesis Screen of Klebsiella pneumoniae Identifies Multiple Genes Important for Resisting Antimicrobial Activities of Neutrophils in Mice

is a Gram-negative bacterial pathogen that causes a range of infections, including pneumonias, urinary tract infections, and septicemia, in otherwise healthy and immunocompromised patients. has become an increasing concern due to the rise and spread of antibiotic-resistant and hypervirulent strains. However, its virulence determinants remain understudied. To identify novel virulence factors needed to cause pneumonia, a high-throughput screen was performed with an arrayed library of over 13,000 transposon insertion mutants in the lungs of wild-type (WT) and neutropenic mice using transposon sequencing (Tn-seq). Insertions in 166 genes resulted in mutants that were significantly less fit in the lungs of WT mice than in those of neutropenic mice. Of these, mutants with insertions in 51 genes still had significant defects in neutropenic mice, while mutants with insertions in 52 genes recovered significantly. screens using a minilibrary of transposon mutants identified putative functions for a subset of these genes, including in capsule content and resistance to reactive oxygen and nitrogen species. Lung infections in mice confirmed roles in virulence for the Δ , Δ , Δ , Δ , Δ , and Δ mutants, all of which were defective in either capsule content or growth in reactive oxygen or nitrogen species. The fitness of the Δ , Δ , Δ , Δ , and Δ mutants was higher in neutropenic mouse lungs, indicating that these genes encode proteins that protect against neutrophil-related effector functions.