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Quantifying the Cytotoxicity of Staphylococcus aureus Against Human Polymorphonuclear Leukocytes
Published on: January 3, 2020
Recognition of apoptotic cells by human peripheral blood monocytes does not alter their ability to phagocytize and
Ewa Zuba1, Kazimierz Weglarczyk, Katarzyna Barczyk
1Department of Immunology, Faculty of Biotechnology, Jagiellonian University, Cracow, Poland.
Introduction:
During acute inflammation, leukocyte infiltration is mostly neutrophilic, but later monocytes prevail. The majority of inflammatory cells, particularly neutrophilic polymorphonuclear leukocytes (PMNs), become apoptotic at later stages of inflammation and are phagocytosed by neighboring cells, mostly by macrophages. Recently, it has been found that human peripheral blood monocytes also recognize apoptotic cells, which primes them to increased production of interleukin (IL)-10--a cytokine known to reduce phagocytes' ability to engulf and kill pathogens. Based on the above, we studied monocytes' ability to phagocytose and kill Staphylococcus aureus while in contact with apoptotic cells.
Materials And Methods:
Monocytes isolated by elutriation were co-cultured with apoptotic PMNs or Jurkat cells and exposed to viable, human serum-opsonized S. aureus. To induce apoptosis PMNs were cultured overnight while Jurkat cells were UV-treated. Apoptosis, phagocytosis of bacteria and intracellular superoxide production were measured by flow cytometry. Production of reactive oxygen species was also followed by measurement of chemiluminescence. The bactericidal effect was determined by standard colony forming units method.
Results:
Data presented show that contact of monocytes with apoptotic neutrophils and Jurkat cells had no influence on monocyte phagocytosis of S. aureus, the generation of reactive oxygen species, or the killing of bacteria.
Conclusion:
The data obtained suggest that monocytes attracted to the inflammatory site are not deficient in their ability to cope with pathogens after contact with apoptotic cells despite increased production of IL-10.
Insights
Contact with apoptotic cells does not impair monocyte function. Monocytes maintain their ability to phagocytose and kill Staphylococcus aureus, even after encountering apoptotic neutrophils and Jurkat cells.
Area of Science:
- Immunology
- Cell Biology
Background:
- Leukocyte infiltration shifts from neutrophils to monocytes during inflammation.
- Apoptotic inflammatory cells, primarily neutrophils, are phagocytosed by macrophages.
- Monocyte exposure to apoptotic cells increases IL-10 production, potentially impairing pathogen defense.
Purpose of the Study:
- To investigate if monocyte interaction with apoptotic cells affects their phagocytosis and killing of Staphylococcus aureus.
- To determine the functional consequences of monocyte-apoptotic cell contact on immune response.
Main Methods:
- Human peripheral blood monocytes were co-cultured with apoptotic neutrophils or Jurkat cells.
- Cells were exposed to opsonized S. aureus.
- Phagocytosis, reactive oxygen species production, and bacterial killing were assessed using flow cytometry and colony-forming unit assays.
Main Results:
- Monocyte phagocytosis of S. aureus was unaffected by co-culture with apoptotic cells.
- Bacterial killing capacity and reactive oxygen species generation remained unchanged.
- Contact with apoptotic neutrophils or Jurkat cells did not impair monocyte antimicrobial functions.
Conclusions:
- Monocytes retain their ability to combat S. aureus following interaction with apoptotic cells.
- Despite potential IL-10 priming, monocytes are not deficient in handling bacterial pathogens at inflammatory sites.
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