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Globally, there are greater than 700,000 deaths per year associated with diarrheal disease. The flagellated intestinal parasite,  Giardia lamblia , is one of the most common intestinal pathogens in both humans and animals throughout the world. While attached to the gastrointestinal epithelium,  Giardia  induces epithelial cell apoptosis, disrupts tight junctions, and increases intestinal permeability. The underlying cellular and molecular mechanisms of giardiasis, including the role lamina propria immune cells, such as macrophages, play in parasite control or clearance are poorly understood. Thus far, one of the major obstacles in ascertaining the mechanisms of  Giardia  pathology is the lack of a functionally relevant model for the long-term study of the parasite  in vitro . Here we report on the development of an  in vitro  co-culture model which maintains the basolateral-apical architecture of the small intestine and allows for long-term survival of the parasite. Using transwell inserts, Caco-2 intestinal epithelial cells and IC-21 macrophages are co-cultured in the presence of  Giardia  trophozoites. Using the developed model, we show that  Giardia  trophozoites survive over 21 days and proliferate in a combination media of Caco-2 cell and  Giardia  medium.  Giardia  induces apoptosis of epithelial cells through caspase-3 activation and macrophages do not abrogate this response. Additionally, macrophages induce Caco-2 cells to secrete the pro-inflammatory cytokines, GRO and IL-8, a response abolished by  Giardia  indicating parasite induced suppression of the host immune response. The co-culture model provides additional complexity and information when compared to a single-cell model. This model will be a valuable tool for answering long-standing questions on host-parasite biology that may lead to discovery of new therapeutic interventions.

Modeling Long-Term Host Cell-Giardia lamblia Interactions in an In Vitro Co-Culture System