Journal
VIRULENCE
Volume 12, Issue 1, Pages 415-429Publisher
TAYLOR & FRANCIS INC
DOI: 10.1080/21505594.2020.1870336
Keywords
Cronobacter sakazakii; lipoprotein; membrane integrity; acid resistance
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Funding
- National Natural Science Foundation of China [31772095]
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Cronobacter sakazakii, an emerging opportunistic pathogen, is associated with severe foodborne outbreak infections in both premature and full-term infants. This study identified the nlpD gene as an acid-resistance factor in C. sakazakii, playing a vital role in maintaining membrane integrity and virulence. The findings suggest that targeting NlpD could be a potential strategy for drug development against C. sakazakii.
Cronobacter sakazakii, an emerging opportunistic pathogen, is implicated in severe foodborne outbreak infections in premature and full-term infants. Generally, acid tolerance is vital for the pathogenesis of foodborne pathogens; however, its role in C. sakazakii virulence remains largely unknown. To screen out acid-tolerance determinants from transposon mutants, anovel counterselection method using gentamicin and acid was developed. Using the counterselection method and growth assay, we screened several acid-sensitive mutants and found that nlpD encodes an acid-resistance factor in C. sakazakii. Compared to the wild-type strain, the nlpD mutant exhibited attenuated virulence in a rat model. Using macrophage THP-1 cells and a pH probe, we verified that nlpD enables bacteria to resist macrophages by resisting acidification. Finally, we confirmed that nlpD maintains C. sakazakii membrane integrity in acid using propidium iodide permeabilization assays via flow cytometry. Our results confirm that nlpD is a novel virulence factor that permits C. sakazakii to survive under acid stress conditions. Considering that NlpD is a conserved lipoprotein located in the bacterial outer membrane, NlpD could be used as a target for drug development.
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