Growing evidence demonstrates a criticalrole of purinergic system in the modulation of the immune response and tissuerepair. Damaged cells release the pro-inflammatory molecule ATP, which is metabolizedby the ectonucleotidases CD39 and CD73 to the anti-inflammatory mediatoradenosine (ADO). Thus, the balance between ATP/ADO is known to determine the outcomeof inflammation/infection. Here, we explored the cellular source of CD73 ectoenzymeexpression in different Trypanosoma cruziinfected tissues and the impact of its activity in local parasitereplication. Through confocal analysis of immunofluorescence assays we foundthat immune cells exhibited significantly higher levels of CD73 expression thanparenchymal cells at 7 days post-infection (dpi), while cardiomyocytes,adipocytes and hepatocytes expressed lower amount of this ectoenzyme (p<0.05immune vs each parenchymal cells). Deficiency of CD73 activity (CD73KO) led to a diminishedcardiac parasite burden, measured by real-time PCR, compared to its wild-type(WT) counterpart (p<0.001 at 21days post-infection(dpi) and p<0.05 at28dpi). Unexpectedly, parasitemia was substantially raised in CD73KO mice incomparison with WT mice (p<0.05 at 14dpi and 21dpi), suggesting theexistence of tissue reservoir/s outside myocardium. Indeed, CD73KO liver andvisceral adipose tissue (VAT) showed increased parasite burden (liver:p<0.001 at 21dpi and VAT: p<0.01 at 21 and 28dpi) associated with areduced ATP/ADO ratio and the lack of substantial microbicidal immune response.These data reveal that the purinergic system has a tissue-dependent impact on parasitereplication and persistence.