title: Establishment of an in vitro system to study intracellular behavior of Candida glabrata in human THP-1 macrophages.
abstract: A cell culture model system, if a close mimic of host environmental conditions, can serve as an inexpensive, reproducible and easily manipulatable alternative to animal model systems for the study of a specific step of microbial pathogen infection. A human monocytic cell line THP-1 which, upon phorbol ester treatment, is differentiated into macrophages, has previously been used to study virulence strategies of many intracellular pathogens including Mycobacterium tuberculosis. Here, we discuss a protocol to enact an in vitro cell culture model system using THP-1 macrophages to delineate the interaction of an opportunistic human yeast pathogen Candida glabrata with host phagocytic cells. This model system is simple, fast, amenable to high-throughput mutant screens, and requires no sophisticated equipment. A typical THP-1 macrophage infection experiment takes approximately 24 hr with an additional 24-48 hr to allow recovered intracellular yeast to grow on rich medium for colony forming unit-based viability analysis. Like other in vitro model systems, a possible limitation of this approach is difficulty in extrapolating the results obtained to a highly complex immune cell circuitry existing in the human host. However, despite this, the current protocol is very useful to elucidate the strategies that a fungal pathogen may employ to evade/counteract antimicrobial response and survive, adapt, and proliferate in the nutrient-poor environment of host immune cells.
authors.affiliation: Laboratory of Fungal Pathogenesis, Centre for DNA Fingerprinting and Diagnostics, Andhra Pradesh, India.
authors.forename: Maruti Nandan
authors.initials: MN
authors.lastname: Rai
authors.forename: Sapan
authors.initials: S
authors.lastname: Borah
authors.forename: Gaurav
authors.initials: G
authors.lastname: Bairwa
authors.forename: Sriram
authors.initials: S
authors.lastname: Balusu
authors.forename: Neelima
authors.initials: N
authors.lastname: Gorityala
authors.forename: Rupinder
authors.initials: R
authors.lastname: Kaur
mesh_terms: D041221:Candida glabrata / Q000276:immunology / Q000472:pathogenicity / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D054884:Host-Pathogen Interactions; D006801:Humans; D007223:Infant; D007948:Leukemia, Monocytic, Acute; D008264:Macrophages / Q000166:cytology* / Q000276:immunology / Q000382:microbiology*; D008297:Male; D009172:Mycology / Q000379:methods*; D014774:Virulence