Clathrin coated pit dependent pathway for Trypanosoma cruzi internalization into host cells.

ABSTRACT

A number of intracellular pathogens are internalized by host cells via multiple endocytic pathways, including Trypanosoma cruzi, the etiological agent of Chagas disease. Clathrin-mediated endocytosis is the most characterized endocytic pathway in mammalian cells. Its machinery was described as being required in mammalian cells for the internalization of large particles, including pathogenic bacteria, fungi, and large virus. To investigate whether T. cruzi entry into host cells can also take advantage of the clathrin-coated vesicle-dependent process, we utilized well-known inhibitors of clathrin-coated vesicle formation (sucrose hypertonic medium, chlorpromazine hydrochloride and pitstop 2) and small interference RNA (siRNA). All treatments drastically reduced the internalization of infective trypomastigotes and amastigotes of T. cruzi by phagocytic (macrophages) and epithelial cells. Clathrin labeling, as observed by fluorescence and electron microscopy, was also observed around the parasites from the initial stages of infection until the complete formation of the parasitophorous vacuole. These unexpected observations suggest the participation of the clathrin pathway in the T. cruzi entry process.