ROS AND POLY(ADP-RIBOSE)POLYMERS GENERATION DURING CELL INVASION BY Trypanosoma cruzi

SALOMÉ C. VILCHEZ LARREA; MARIA LAURA KEVORKIAN; MARIANA SCHLESINGER; MIRTHA M. FLAWIA; SILVIA FERNANDEZ VILLAMIL

Congreso; X Congreso de Protozoología y Enfermedades parasitarias; 2014

Sociedad Argentina de Protozoologia y Enfermedades Parasitarias

Poly (ADP-ribose) (pADPr) metabolism has been demonstrated to be important for the normal progression of the infection cycle of T. cruzi in different host cell lines. Based on previous results obtained by our group and others, it is possible to hypothesize that cell invasion by T. cruzi leads to the production of reactive oxygen species (ROS) and the trigger of an inflammatory response. Generated ROS also cause DNA damage, which would in turn activate PARP-1, NFkB and cytokine expression. Up to date, we have proven that infection levels in Vero cells is diminished by the presence of Olaparib, a PARP-1 inhibitor, or DEA, a PARG inhibitor and that this is observed when using two different T. cruzi strains: CL Brener, of low infectivity, and Tulahuen II, a more aggressive strain. The use of a more specific PARG inhibitor, RBPI-5, corroborated the result obtained, indicating that the observed outcome is due to the specific absence of PARG activity. Using fluorescent probes, the formation of ROS in the cytoplasm of the cell after infection by trypomastigotes was seen and the generation of pADPr, both in the nuclei of intracellular amastigotes and in the cytoplasm of the host cell in the vicinity of these replicative forms, could also be detected, probably as a response to the action of ROS. Nevertheless, both PARP-1 and PARG remained in the nucleus of the host cell during the infection process. This could indicate that the cytoplasmatic pADPr observed would be synthesized by PARPs other than PARP-1, being Tankyrase a good candidate for this.