Steinhaeuser, Christine and Heigl, Ulrike and Tchikov, Vladimir and Schwarz, Jeanette and Gutsmann, Thomas and Seeger, Katrin and Brandenburg, Julius and Fritsch, Juergen and Schroeder, Josef and Wiesmueller, Karl-Heinz and Rosenkrands, Ida and Walther, Paul and Pott, Johanna and Krause, Eberhard and Ehlers, Stefan and Schneider-Brachert, Wulf and Schuetze, Stefan and Reiling, Norbert (2013) Lipid-Labeling Facilitates a Novel Magnetic Isolation Procedure to Characterize Pathogen-Containing Phagosomes. TRAFFIC, 14 (3). pp. 321-336. ISSN 1398-9219,
Full text not available from this repository. (Request a copy)Abstract
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 Mycobacterium avium, M. tuberculosis and Listeria monocytogenes and isolated magnetic bacteria-containing phagosomes using a strong magnetic field in a novel free-flow system. Magnetic labeling of M.tuberculosis did not affect the virulence characteristics of the bacteria during infection experiments addressing host cell activation, phagosome maturation delay and replication in macrophages in vitro. 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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MYCOBACTERIUM-TUBERCULOSIS PHAGOSOME; 2-DIMENSIONAL GEL-ELECTROPHORESIS; NECROSIS-FACTOR-ALPHA; INTERFERON-GAMMA; ACTIVATED MACROPHAGES; LYSOSOMAL PROTEOLYSIS; CULTURED MACROPHAGES; ELECTRON-MICROSCOPY; NITRIC-OXIDE; MATURATION; Gram-positive bacteria; infection; macrophages; magnetic isolation; mycobacteria; phagosomes; primary cells; tuberculosis |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 24 Apr 2020 04:49 |
| Last Modified: | 24 Apr 2020 04:49 |
| URI: | https://pred.uni-regensburg.de/id/eprint/17087 |
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