Assessing the effect of antimony pressure on trypanothione reductase activity in Leishmania (Viannia) braziliensis

Abstract

Leishmania parasites have an oxidative and chemical defense mechanism called trypanothione system (T[SH]₂), the most abundant thiol system in trypanosomatids. This system has a central role in processing pentavalent antimony and resistance has been related to a better capacity to metabolize it through the activation of T[SH]₂ enzymatic cascade. A biochemical approach was applied to assess the effect of trivalent (Sb^III) and pentavalent antimony (Sb^V) on Trypanothione Reductase (TR) activity of two Leishmania (Viannia) braziliensis clinical isolates, which were labeled as responder (R) and non-responder (NR) after patient treatment with Glucantime®. Both isolates were characterized based on in vitro susceptibility to Sb^III and Sb^V and trypanothione reductase (TR) activity. Sb^III and Sb^V discriminated susceptibility profiles in all parasite forms, since isolate NR had significantly higher EC₅₀ values than isolate R. Differences were observed in TR activity between promastigotes, axenic amastigotes and intracellular amastigotes: R (0.439 ± 0.009, 0.103 ± 0.01 and 0.185 ± 0.01AU.min^-1.μg of protein^-1) and NR (1.083 ± 0.04, 0.914 ± 0.04 and 0.343 ± 0.04 AU. min^-1.μg of protein^-1), respectively. Incubation with Sb^III and Sb^V using each form EC₅₀ value caused a time-dependent differential effect on TR activity suggesting that oxidative defense is related to the antimony susceptibility phenotype. Data gathered here shows a biochemical approach able to discriminate two L. (V.) braziliensis clinical isolates measurements TR activity of promastigotes, axenic amastigotes and intracellular amastigotes.

Keywords: Antimony; Clinical isolates; Leishmania (Viannia) braziliensis; Trypanothione reductase.