Lipid‐Labeling Facilitates a Novel Magnetic Isolation Procedure to Characterize Pathogen‐Containing Phagosomes. (28th December 2012)

• Record Type: Journal Article
• Authors: Steinhäuser, Christine; Heigl, Ulrike; Tchikov, Vladimir; Schwarz, Jeanette; Gutsmann, Thomas; Seeger, Katrin; Brandenburg, Julius; Fritsch, Jürgen; Schroeder, Josef; Wiesmüller, Karl‐Heinz; Rosenkrands, Ida; Walther, Paul; Pott, Johanna; Krause, Eberhard; Ehlers, Stefan; Schneider‐Brachert, Wulf; Schütze, Stefan; Reiling, Norbert
• Is Part Of: Traffic. Volume 14:Number 3(2013:Mar.)
• Page Start: 321
• Page End: 336
• Publication Date: 2012-12-28
• Subjects: Biological transport -- Periodicals; 571.6
• DOI: 10.1111/tra.12031
• Languages: English
• ISSNs: 1398-9219

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p> <bold>Here we describe a novel approach for the isolation and biochemical characterization of pathogen‐containing compartments from primary cells: We developed a lipid‐based procedure to magnetically label the surface of bacteria and visualized the label by scanning and transmission electron microscopy (SEM, TEM). We performed infection experiments with magnetically labeled <italic>Mycobacterium avium</italic>, <italic>M. tuberculosis</italic> and <italic>Listeria monocytogenes</italic> and isolated magnetic bacteria‐containing phagosomes using a strong magnetic field in a novel free‐flow system. Magnetic labeling of <italic>M. tuberculosis</italic> did not affect the virulence characteristics of the bacteria during infection experiments addressing host cell activation, phagosome maturation delay and replication in macrophages <italic>in vitro</italic>. Biochemical analyses of the magnetic phagosome‐containing fractions provided evidence of an enhanced presence of bacterial antigens and a differential distribution of proteins involved in the endocytic pathway over time as well as cytokine‐dependent changes in the phagosomal protein composition. The newly developed method represents a useful approach to characterize and compare pathogen‐containing compartments, in order to identify microbial and host cell targets for novel anti‐infective strategies.</bold> </p> </abstract>