Visceral leishmaniasis (VL) infection is mostly caused by Leishmania donovani and affects countries worldwide. Despite the need for a safe and effective vaccine against leishmaniasis due to the increased drug resistance, however, no vaccine has yet been licensed for clinical use. This study revolves around the immunoinformatics approach to design a multi-epitope vaccine against VL infection. In this case, the proteome of L. donovani has been investigated, and three host non-homologous and antigenic extracellular secretory proteins have been identified as potential vaccine candidates with low transmembrane helices (≤ 1). The multi-epitope subunit vaccine construct consists of T-cell (cytotoxic T-lymphocyte (CTL) and helper T-lymphocyte (HTL)) epitopes accompanied by appropriate adjuvant and linkers. A 372-amino acid vaccine construct has been established with specific characteristics, such as soluble, stable, antigenic, non-allergenic, non-toxic, and non-host homologous. Besides, the tertiary structure of the designed vaccine was modeled and validated. Also, the stability and affinity of the vaccine- TLR4 complex were confirmed by using molecular docking and molecular dynamics (MD) simulation. In addition, in silico immunization assay showed the efficiency of this candidate vaccine to stimulate an effective immune response. Furthermore, the refined vaccine was optimized and cloned in the pET28a (+) vector, and its successful expression was confirmed virtually. However, the experimental validation is required to verify the multi-epitope vaccine efficacy against VL infection.

Background:The current study was conducted to compare ELISA with thin layer

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