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Updated: Jul 4, 2026
Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages
Published on: October 25, 2017
Isolation of bacteria-containing phagosomes by magnetic selection
Per Lönnbro1, Pontus Nordenfelt, Hans Tapper
1Division of Infection Medicine, Department of Clinical Sciences, Lund University, SE-221 84 Lund, Sweden. per.lonnbro@med.lu.se
Background:
There is a growing awareness of the importance of intracellular events in determining the outcome of infectious disease. To improve the understanding of such events, like phagosome maturation, we set out to develop a versatile technique for phagosome isolation that is rapid and widely applicable to different pathogens.
Results:
We developed two different protocols to isolate phagosomes containing dead or live bacteria modified with small magnetic particles, in conjunction with a synchronized phagocytosis protocol and nitrogen cavitation. For dead bacteria, we performed analysis of the phagosome samples by microscopy and immunoblot, and demonstrated the appearance of maturation markers on isolated phagosomes.
Conclusion:
We have presented detailed protocols for phagosome isolation, which can be adapted for use with different cell types and prey. The versatility and simplicity of the approach allow better control of phagosome isolation, the parameters of which are critical in studies of host-bacteria interaction and phagosome maturation.
Insights
Researchers developed a versatile magnetic particle-based method for isolating phagosomes, crucial for studying host-bacteria interactions and phagosome maturation in infectious diseases.
Area of Science:
- Immunology
- Cell Biology
- Microbiology
Background:
- Intracellular events significantly influence infectious disease outcomes.
- Phagosome maturation is a key intracellular event requiring further study.
- A versatile and rapid phagosome isolation technique is needed.
Purpose of the Study:
- To develop a versatile, rapid, and widely applicable technique for phagosome isolation.
- To facilitate a better understanding of intracellular events in infectious diseases.
Main Methods:
- Developed two protocols for phagosome isolation using magnetic particles.
- Employed synchronized phagocytosis and nitrogen cavitation.
- Adapted for isolating phagosomes containing live or dead bacteria.
Main Results:
- Successfully isolated phagosomes containing modified bacteria.
- Demonstrated the appearance of maturation markers on isolated phagosomes (for dead bacteria) via microscopy and immunoblot.
- Validated the technique's applicability to different bacterial states.
Conclusions:
- Presented detailed, adaptable protocols for phagosome isolation.
- The method offers versatility and simplicity for controlling isolation parameters.
- Enables improved studies of host-bacteria interactions and phagosome maturation.
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