SKAP2 is required for defense against K. pneumoniae infection and neutrophil respiratory burst

Elife. 2020 Apr 30;9:e56656. doi: 10.7554/eLife.56656.

Abstract

Klebsiella pneumoniae is a respiratory, blood, liver, and bladder pathogen of significant clinical concern. We show that the adaptor protein, SKAP2, is required for protection against K. pneumoniae (ATCC 43816) pulmonary infections. Skap2-/- mice had 100-fold higher bacterial burden when compared to wild-type and burden was controlled by SKAP2 expression in innate immune cells. Skap2-/- neutrophils and monocytes were present in infected lungs, and the neutrophils degranulated normally in response to K. pneumoniae infection in mice; however, K. pneumoniae-stimulated reactive oxygen species (ROS) production in vitro was abolished. K. pneumoniae-induced neutrophil ROS response required the activity of SFKs, Syk, Btk, PLCγ2, and PKC. The loss of SKAP2 significantly hindered the K. pneumoniae-induced phosphorylation of SFKs, Syk, and Pyk2 implicating SKAP2 as proximal to their activation in pathogen-signaling pathways. In conclusion, SKAP2-dependent signaling in neutrophils is essential for K. pneumoniae-activated ROS production and for promoting bacterial clearance during infection.

Keywords: Hoxb8; Klebsiella pneumoniae; SKAP2; bacterial pneumonia; immunology; infectious disease; inflammation; microbiology; mouse; neutrophils; reactive oxygen species.

Plain Language Summary

Klebsiella pneumoniae is a type of bacteria that can cause life-threatening infections – including pneumonia, blood stream infections, and urinary tract infections – in hospitalized patients. These infections can be difficult to treat because some K. pneumoniae are resistant to antibiotics. The bacteria are normally found in the human intestine, and they do not usually cause infections in healthy people. This implies that healthy people’s immune systems are better able to fend off K. pneumoniae infections; learning how could help scientists develop new ways to treat or prevent infections in hospitalized patients. In healthy people, a type of immune cell called neutrophils are the first line of defense against bacterial infections. Several different proteins are needed to activate neutrophils, including a protein called SKAP2. But the role of this protein in fighting K. pneumoniae infections is not clear. To find out what role SKAP2 plays in the defense against pneumonia caused by K. pneumoniae, Nguyen et al. compared infections in mice with and without the protein. Mice lacking SKAP2 in their white blood cells had more bacteria in their lungs than normal mice. The experiments showed that neutrophils from mice with SKAP2 produce a burst of chemicals called “reactive oxygen species”, which can kill bacteria. But neutrophils without the protein do not. Without SKAP2, several proteins that help produce reactive oxygen species do not work. Understanding the role of SKAP2 in fighting infections may help scientists better understand the immune system. This could help clinicians to treat conditions that cause it to be hyperactive or ineffective. More studies are needed to determine if SKAP2 works the same way in human neutrophils and if it works against all types of K. pneumoniae. If it does, then scientists might be able use this information to develop therapies that help the immune system fight infections.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bacterial Load
  • Cell Line
  • Disease Models, Animal
  • Focal Adhesion Kinase 2 / metabolism
  • Intracellular Signaling Peptides and Proteins / deficiency
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Klebsiella Infections / genetics
  • Klebsiella Infections / immunology
  • Klebsiella Infections / metabolism*
  • Klebsiella Infections / microbiology
  • Klebsiella pneumoniae / pathogenicity*
  • Lung / immunology
  • Lung / metabolism*
  • Lung / microbiology
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Neutrophils / immunology
  • Neutrophils / metabolism*
  • Neutrophils / microbiology
  • Phosphorylation
  • Pneumonia, Bacterial / genetics
  • Pneumonia, Bacterial / immunology
  • Pneumonia, Bacterial / metabolism*
  • Pneumonia, Bacterial / microbiology
  • Reactive Oxygen Species / metabolism
  • Respiratory Burst*
  • Signal Transduction
  • Syk Kinase / metabolism

Substances

  • Intracellular Signaling Peptides and Proteins
  • Reactive Oxygen Species
  • src kinase associated phosphoprotein 2
  • Focal Adhesion Kinase 2
  • Ptk2b protein, mouse
  • Syk Kinase
  • Syk protein, mouse