Destruction ofLeishmania mexicana amazonensis amastigotesby leucine methyl ester: protection by other amino acid esters

Abstract

SUMMARY: l-Amino acid esters, such as leucine methyl ester (Leu-OMe) can destroy intracellular as well as isolated amastigotesof Leishmania mexicana amazonensisby a mechanism which may involve ester hydrolysis by parasite enzymes. We show here that several other esters prevented the killing of the amastigotes by Leu-OMe. Destruction ofLeishmaniawithin macrophages in culture was assessed microscopically and viability of isolated parasites was monitored by reduction of the tetrazolium MTT. The main features of the protective effect were similar for intracellular and for isolated amastigotes. Thus, (i) effective prevention of parasite killing required that the protective ester be present in the medium prior to and during exposure of infected cells or parasites to Leu-OMe; (ii) the same esters protected intracellular and isolatedLeishmaniaagainst damage by Leu-OMe. Ranks of protective activity, as determined on isolated amastigotes were: Gly-OBz > Tyr-OMe > Ile-OMe > Met-OMe > Val-OMe > Ala-OMe > Gly-OMe > D-Leu-OMe; (iii) several esters were inactive in both systems (Leu-OBz, Trp-OMe and Phe-OMe). Protective activity was associated with leishmanicidal (e.g. Gly-OBz, Tyr-OMe) as well as with non-leishmanicidal (e.g. Ile-OMe, Val-OMe) esters. The results are compatible with the hypothesis that protective esters inhibit the activity of parasite enzyme(s) which hydrolyse Leu-OMe.