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Leishmaniasis is a significant worldwide health problem for which no vaccine exists. Activation of CD4 +  and CD8 +  T cells is crucial for the generation of protective immunity against parasite. Recent trend in vaccine design has been shifted to epitope-based vaccines that are more specific, safe, and easy to produce. In the present study, four known antigenic  Leishmania infantum  proteins, cysteine peptidase A (CPA), histone H1, KMP-11, and  Leishmania  eukaryotic initiation factor (LeIF) were analyzed for the prediction of binding epitopes to H2 d  MHC class I and II molecules, using online available algorithms. Based on  in silico  analysis, eight peptides including highly scored MHC class I- and II-restricted epitopes were synthesized. Peptide immunogenicity was validated in MHC compatible BALB/c mice immunized with each synthetic peptide emulsified in complete Freund’s adjuvant/incomplete Freund’s adjuvant. CPA_p2, CPA_p3, H1_p1, and LeIF_p6 induced strong spleen cell proliferation upon  in vitro  peptide re-stimulation. In addition, the majority of the peptides, except of LeIF_p1 and KMP-11_p1, induced IFN-γ secretion, while KMP-11_p1 indicated a suppressive effect on IL-10 production. CPA_p2, CPA_p3, LeIF_p3, and LeIF_p6 induced IFN-γ-producing CD4 +  T cells indicating a T H1 -type response. In addition, CPA_p2, CPA_p3, and H1_p1 induced also the induction of CD8 +  T cells. The induction of peptide-specific IgG in immunized mice designated also the existence of B cell epitopes in peptide sequences. Combining immunoinformatic tools and experimental validation, we demonstrated that CPA_p2, CPA_p3, H1_p1, H1_p3, CPA_p2, LeIF_p3, and LeIF_p6 are likely to include potential epitopes for the induction of protective cytotoxic and/or T H1 -type immune responses supporting the feasibility of peptide-based vaccine development for leishmaniasis.

frontiers in immunology

Experimental Validation of Multi-Epitope Peptides Including Promising MHC Class I- and II-Restricted Epitopes of Four Known Leishmania infantum Proteins