Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 16 2 2021 05 28 186 198 Niloofar Bavarsad Ahmadpour Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Abdolhossein Dalimi Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Majid Pirestani Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Javid Sadraei Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2020 03 02 2021 05 28 Background: Leishmania is a mandatory intracellular pathogen and causing neglected disease. Hence, protection against leishmaniasis by a development vaccine is an important subject. This study aimed to design a poly-epitope vaccine for cutaneous leishmaniasis. Methods: The present study was conducted in the Parasitology Department of Tarbiat Modares University, Tehran, Iran during 2017-2019. Several bioinformatics methods at online servers were used for prediction of different aspects of poly-epitope, including, physico-chemical attributes, allergenicity, antigenicity, secondary and tertiary structures, B-cell, T-cell and MHC (I, II) potential epitopes of LACK, LEIF, GP63 and SMT antigens of L. major. Results: After designing the construct (GLSL), the outputs of PTM sites demonstrated that the poly-epitope had 57 potential sites for phosphorylation. Furthermore, the secondary of GLSL structure includes 59.42%, 20.94% and 19.63% for random coil, extended strand and alpha-helix, respectively. The GLSL is an immunogenic protein with an acceptable antigenicity (0.8410) and non-allergen. Afterward, 20 potential epitopes of LACK, LEIF, GP63 and SMT antigens were linked by a flexible linker (SAPGTP), then was synthesized, and sub-cloned in pLEXY– neo2. The results were confirmed the expression of 38.7 kDa poly-epitope in secretory and cytosolic sites, separately. Conclusion: A good expression in the L. tarentulae and confirmation of the GLSL poly-epitope could be a basis for developing a vaccine candidate against leishmaniasis that should be confirmed via experimental tests in BALB/c mice. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/2816 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/2816/1123