Abstract
Two Leishmania infantum mimotopes (B10 and C01) identified by phage display showed to be antigenic and immunogenic for visceral (VL) and tegumentary (TL) leishmaniasis; however, their biological targets in the parasites have not been identified. The aim of the present study was to investigate the native antigens expressing both mimotopes, and to use them in distinct immunological assays. For this, a subtractive phage display technology was used, where a combinatorial library of single-chain variable fragments (scFv) was employed and the most reactive monoclonal antibodies for each target were captured, being the target antigens identified by mass spectrometry. Results in immunoblotting and immunoprecipitation assays showed that both monoclonal scFvs antibodies identified the β-tubulin protein as the target antigen in L. infantum. To validate these findings, the recombinant protein was cloned, purified and tested for the serodiagnosis of human leishmaniasis, and its immunogenicity was evaluated in PBMC derived from healthy subjects and treated or untreated VL patients. Results showed high diagnostic efficacy, as well as the development of a specific Th1 immune response in the cell cultures, since higher IFN-γ and lower IL-10 production was found.
Highlights
Leishmaniasis is an infectious disease caused by protozoan parasites of the genus Leishmania, through the bite of infected phlebotomine sand flies [1]
There is no an available vaccine against human visceral leishmaniasis (VL), some candidates have been well-evaluated in experimental trials using mice and dogs [5]
After the bio-selection cycles, clones were grown in 96 deep-well plates, and expression induction of scFv antibodies was evaluated by ELISA
Summary
Leishmaniasis is an infectious disease caused by protozoan parasites of the genus Leishmania, through the bite of infected phlebotomine sand flies [1]. Brazil is responsible by about 90% of VL cases, with the L. infantum species being the main parasite species responsible for the disease in dogs and humans [3]. Prophylactic measures have not been efficient to control this disease, and the available antileishmanial drugs present problems regarding their toxicity and/or high cost [4]. The development of an effective vaccine could be considered as a strategy to control the spread of the disease. There is no an available vaccine against human VL, some candidates have been well-evaluated in experimental trials using mice and dogs [5]
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.
