Leishmania are obligate intracellular parasites known to have developed successful ways of efficient immunity evasion. Because of this, leishmaniasis, a disease caused by these flagellated protists, is ranked as one of the most serious tropical infections worldwide. Neither prophylactic medication, nor vaccination has been developed thus far, even though the infection has usually led to strong and long-lasting immunity. In this paper, we describe a "suicidal" system established in Leishmaniamexicana, a human pathogen causing cutaneous leishmaniasis. This system is based on the expression and (de)stabilization of a basic phospholipase A2 toxin from the Bothropspauloensis snake venom, which leads to the inducible cell death of the parasites in vitro. Furthermore, the suicidal strain was highly attenuated during macrophage infection, regardless of the toxin stabilization. Such a deliberately weakened parasite could be used to vaccinate the host, as its viability is regulated by the toxin stabilization, causing a profoundly reduced pathogenesis.

• Publication type
• Journal Article MeSH

Targeted regulation of protein levels is an important tool to investigate the role of proteins essential for cell function and development. In recent years, methods based on the Escherichia coli dihydrofolate reductase destabilization domain (ecDHFR DD) have been established and used in various cell types. ecDHFR DD destabilizes the fused protein of interest and causes its degradation by proteasomes, unless it is stabilized by a specific ligand, trimethoprim. In this work we developed an inducible protein stabilization system in Leishmania mexicana based on ecDHFR DD.

• MeSH
• Transcriptional Activation * MeSH
• Tetrahydrofolate Dehydrogenase genetics   metabolism   MeSH
• Escherichia coli enzymology   genetics   MeSH
• Leishmania mexicana genetics   metabolism   MeSH
• Molecular Biology methods   MeSH
• Parasitology methods   MeSH
• Gene Expression Regulation * MeSH
• Recombinant Proteins genetics   metabolism   MeSH
• Trimethoprim metabolism   MeSH
• Publication type
• Journal Article MeSH