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Abstract

FURTHER STUDIES OF DEATH AND PROGRESSIVE ELIMINATION OF INTRACELLULAR TOXOPLASMA GONDII IN PRESENCE OF HYDROXYUREA

Edésio José Tenório of Melo*, Laís Pessanha de Carvalho and Cristiane de Souza Carvalho Wodarz

ABSTRACT

Toxoplasma gondii tachyzoite infects and resides within a special parasitophorous vacuole in eukaryotic cells. Viable parasites manipulate the vacuole allowing the uptake of nutrients from the host cell while avoid parasitophorous vacuole fusion with the host cell endocytic pathway. We have shown that hydroxyurea arrests parasite multiplication, leading to parasitophorous vacuole acidification and elimination of the parasite. However, cellular and kinetic events associated with this were not described. In the present study we investigate the successive steps of Toxoplasma destruction in presence of hydroxyurea. For these purposes, Vero cells were infected with tachyzoite of Toxoplasma gondii and treated with increasing amounts of hydroxyurea for several times. We showed that the arrest of tachyzoite multiplication occurred after 5 hours of treatment, coinciding with the time required for parasite division cycle. After 8 hours of incubation, treated cultures had amorphous parasites-containing parasitophorous vacuole, which fused with host cell lysosome. After 12 h, disrupted parasites-containing autophagic vacuoles were also visible. After 24 h, parasite DNA fragmentation also occurred. Once the tachyzoites had been eliminated, treated cells recovered mitochondrial functionality and distribution on the host cell cytoplasm. These assays suggested that the main target of hydroxyurea is the intravacuolar proliferative tachyzoites and host cells recovered their defense against intracellular T. gondii during hydroxyurea treatment. The observations reported in this paper also imply that hydroxyurea would be a useful tool in the study of host–pathogen interactions, mainly in the study of Apicomplexa cell biology and in the development of more specific therapeutic agents.

Keywords: apoptosis, cell cycle, hydroxyurea, parasitophorous vacuole-lysosome fusion, Toxoplasma gondii.


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