Detection and quantification of microbial manipulation of phagosomal function

Abstract

Macrophages are phagocytic cells that constitute the primary barrier against microbial infection. However, the frequency of this interaction has likely led to the selection and retention of specific strategies to subvert the antimicrobial behavior of these cells. Many intracellular pathogens manipulate their host macrophages, in order to survive, avoid degradation and immune recognition. Much of the antimicrobial defenses mobilized by the macrophage function through the lysosomal compartment and it is the subversion of these mechanisms that are the target of many pathogenic microbes. To investigate the potential manipulation of lysosomal function by infectious agents, the protocols provided here describe a wide range of techniques developed to study various lysosome functions, ranging from phagosome-lysosome fusion to a panel of hydrolytic activities. The suitability of various protocols for specific contexts is addressed. In addition to providing detailed methods for the elucidation of lysosome function and their applications to microbial infections, approaches are also discussed for developing new protocols that would complement our knowledge of microbial manipulation of lysosome functions. The continued technical evolution of these methods is central to increasing our understanding of host-pathogen interactions and cell biology in general.

Keywords: Fluorogenic sensors; Intracellular microbes; Lysosome function; Macrophage; Phagosome maturation.