Stochastic induction of Theileria annulata merogony in vitro by chloramphenicol

Theileria annulata inhabits the cytoplasm of bovine leukocytes where it can be found as a multinucleated schizont. The schizont is the pathogenic stage of the life cycle and by interfering with host signalling pathways, it induces unlimited host cell proliferation and protection against apoptosis. In the infected animal, the schizont differentiates to the merozoite life cycle stage in a process called merogony. This takes place within the host leukocyte, resulting in the production of merozoites that are subsequently released by leukocyte lysis. In established cultures of T. annulata-transformed cells, merogony does not spontaneously occur, but the process can be activated by a shift in temperature. In this study we show that chloramphenicol induces schizont differentiation in proliferating T. annulata-transformed cells. We demonstrate that chloramphenicol-induced merogony is inherently asynchronous and has a quantitative basis. The process is accompanied by the down-regulation of schizont-specific surface proteins, de novo expression of merozoite-specific markers such as Tamr1 and Tams1 and the morphological hallmarks of merogony. Chloramphenicol-induced parasite differentiation was found to be associated with diminished proliferation potential and extensive morphological changes of the host cell, including increased numbers of pseudopodia. Significantly, chloramphenicol treatment can accelerate merogony induced by elevated temperature, supporting postulation that the differentiation event is a stochastic process that can be manipulated to alter the outcome of parasitic infection.