Rab32 and Rab9 are present in the trypanosoma cruzi parasitophorous vacuole and contribute to the infection process

SALASSA BN; CUETO JA; VANRELL MC; MARTINEZ SJ; FICHELE MM; ROMANO PS

Congreso; Congrès Armand-Frappier 2019; 2019

Trypanosoma cruzi, the etiological agent of Chagas disease is a protozoan parasite which infects both phagocytic and non-phagocytic mammalian cells. At early stages of infection, trypomastigotes localize in a vesicular compartment called the T. cruzi parasitophorous vacuole (TcPV) until the escape to cell cytoplasm to continue their cycle. Rab and SNAREs proteins take part of the molecular machinery of membrane traffic. Rab GTPases have emerged as central regulators of vesicle recognition and transport, whereas SNAREs are mainly involved in the fusion process between membranes.In a previous work, we proposed that the T. cruzi infection process in non-phagocytic cells occurs in two stages, the formation and the maturation of the TcPV. Indeed, we showed that VAMP7 is required for the TcPV development and for the establishment of infection. The aim of this work was to identify other molecular components of the vesicular transport pathways and their participation in the T. cruzi infection. CHO cells were transfected to overexpress GFP-Rab proteins and infected with trypomastigotes for different times. Then, cells were fixed and processed to detect intracellular parasites by indirect immunofluorescence. Similar procedures were performed in CHO cells overexpressing GFP-vector as control. By confocal microscopy, we observed that Rab9 was recruited to the membrane of the TcPV during both stages: formation and maturation of the TcPV. Likewise, we detected that Rab32 was also recruited to TcPV during its maturation, in parallel with VAMP7. Interestingly, the recruitment to TcPV of both, Rab9 and Rab32, was specific of T. cruzi due to it was not occurred on latex beads or when parasite protein synthesis was inhibited. In addition, we found that the overexpression of Rab32 significantly increased T. cruzi infection. To this moment we concluded that both Rab9 and Rab32 are important for the intracellular cycle of T. cruzi favoring the formation and /or the maturation of the TcPV that results in an increased infection.