AbstractThe gram‐negative bacterium Coxiella burnetii causes human Q fever, a disease characterized by a debilitating flu‐like illness in acute cases and endocarditis in chronic patients. An obligate intracellular pathogen, Coxiella burnetii survives within a large, lysosome‐like vacuole inside the host cell. A unique feature of the Coxiella parasitophorous vacuole (PV) is high levels of fusion with the host endocytic pathway, with PV‐endosome fusion critical for Coxiella survival within the host cell. This unit describes quantitating PV‐endosome fusion by measuring delivery of the fluid phase endosome marker dextran to the PV using live cell imaging. To study the effect of host cell proteins involved in PV‐endosome fusion, details are provided for using siRNA knockdown host cells. This method is a powerful tool for understanding mechanisms underlying Coxiella’s ability to manipulate host cell trafficking pathways. © 2017 by John Wiley & Sons, Inc.
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