Isolation of bacteria-containing phagosomes by magnetic selection.
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.


CONCLUSIONS
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.
Lönnbro
Per
P
Division of Infection Medicine, Department of Clinical Sciences, Lund University, SE-221 84 Lund, Sweden. per.lonnbro@med.lu.se
Nordenfelt
Pontus
P
Tapper
Hans
H
D001431:Bacteriological Techniques; D035843:Biomedical Research / Q000379:methods*; D002458:Cell Fractionation / Q000379:methods*; D016130:Immunophenotyping; D008280:Magnetics; D010588:Phagosomes / Q000382:microbiology*