Abstract
An immunostimulatory glycolipid molecule from the intestinal protozoan parasite Entamoeba histolytica (Eh) and its synthetic analogs derived from its phosphatidylinositol-b-anchor (EhPIb) previously showed considerable immunotherapeutic effects against Leishmania major infection in vitro and in vivo. Here, we describe a high content screening assay, based on primary murine macrophages. Parasites detection is based on a 90 kDA heat shock protein-specific staining, enabling the detection of several Leishmania species. We validated the assay using L. major, L. braziliensis, L. donovani, and L. infantum as well as investigated the anti-leishmanial activity of six immunostimulatory EhPIb-compounds (Eh-1 to Eh-6). Macrophages infected with dermotropic species were more sensitive towards treatment with the compounds as their viability showed a stronger reduction compared to macrophages infected with viscerotropic species. Most compounds caused a significant reduction of the infection rates and the parasite burdens depending on the infecting species. Only compound Eh-6 was found to have activity against all Leishmania species. Considering the challenges in anti-leishmanial drug discovery, we developed a multi-species screening assay capable of utilizing non-recombinant parasite strains, and demonstrated its usefulness by screening macrophage-targeting EhPIb-compounds showing their potential for the treatment of cutaneous and visceral leishmaniasis.
Highlights
We recently reported the immunostimulatory activity of a novel set of synthetic analogs derived from the phosphatidylinositol b anchor (PIb) of a lipopeptidephosphoglycan (LPPG) isolated from the membrane of the protozoan parasite Entamoeba histolytica (Eh)
We demonstrated its usefulness by screening the anti-leishmanial activity of a set of synthesized immunostimulatory EhPIb-compounds against intracellular amastigotes of both dermotropic and viscerotropic Leishmania species
At 24 h post infection, immunostimulatory EhPIb-compounds were added to the cell cultures for 24 and 48 h followed by fixation, immunofluorescent staining, and image acquisition using the Opera Phenix high content screening (HCS) system (Figure 1A)
Summary
The leishmaniases are an increasingly prevalent, poverty-related, and complex group of neglected diseases caused by infection with obligate intracellular parasites of the genus. 20 Leishmania (L.) species are pathogenic to humans and are transmitted by the bite of female sandflies, e.g., Phlebotomus spp. and Lutzomyia spp. Leishmaniasis affects over 12 million people worldwide and is endemic in at least 98 tropical and subtropical countries with an estimated number of 700,000 to 1 million new cases annually [1]. Clinical symptoms can vary greatly, depending on the infecting species and host immunological factors, ranging from mild self-limiting cutaneous ulcers (cutaneous 4.0/).
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
