Mechanism of entry and development of Trypanosoma dionisii in non-phagocytic cells

Cultures of 'buffalo' (bison) lung (BL) cells were infected with epimastigotes or trypomastigotes of Trypanosoma (Schizotrypanum) dionisii derived from cultures in vitro, fixed after various periods of incubation at 37 degrees C and examined by light or electron microscopy. Few if any epimastigotes entered the BL cells, but many trypomastigotes did so; they adhered to the cell surface within 2 h and then appeared to sink into furrows on the cell surface until engulfed in parasitophorous vacuoles. Cytochalasin D (5-10 micrograms ml-1) completely, but reversibly, inhibited entry of trypomastigotes without affecting parasite motility. It was concluded that entry depended on the interaction of stage-specific components on the trypomastigote's surface with receptors on the BL cells, and that this interaction induced active uptake of the protozoa by a phagocytic process not involving pseudopod formation. Soon after entry of the trypomastigotes into BL cells, the membranes of the parasitophorous vacuoles disintegrated and the parasites, which were now lying free in the cytoplasm of the host cell, transformed into amastigotes (micromastigotes) during the next 24-48 h. Replication then occurred, followed by transformation, beginning after 3 days, through a transitional promastigote phase to small intracellular trypomastigotes at 7 days. The promastigotes had a characteristic curved protrusion extending from the lip of the flagellar pocket (or reservoir) into the host cell's cytoplasm. Trypomastigotes, released into the supernatant medium by rupture of the plasma membranes of the BL cells after 8 days, could re-invade other cells.