In vivo study of inactive trans-sialidase expression in naturally lacking parasites

JUAN BURGOS; MARÍA S. LEGUIZAMÓN; MIRIAM POSTAM; CARLA A. PASCUALE; OSCAR CAMPETELLA

Congreso; Reunión conjunta de sociedades de biociencias; 2017

Trypanosoma cruzi trans-sialidase (TS) has arised as a relevant parasite virulence factor. TS family contain active (aTS) and inactive (iTS) isoforms with a His342Tyr conserved mutation. We have reported that, in contrast to high-virulence strains (DTUs II, V and VI), lower-virulence strains (DTUs I, III and IV) do not contain iTS encoding genes. Previous in vitro results using recombinant iTS suggest that iTS could play a different or complementary pathogenic role to that of aTS protein. We have constructed a recombinant iTS-encoding gene where a 3XFLAG epitope was inserted (T-iTS) and used to transfect null-parasites (K98 DTU I). Flow Citometry, Confocal Microscopy, Western Blot and TS activity evaluation showed the adecuate expression and localization. Our in vitro assay showed that T-iTS transfected clones were able, at low interaction time of interaction (1 h), to infect significant more cells (Vero, Cardiocytes and fibroblast, p