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Background: Hepatocellular carcinoma (HCC) is one of the prevalent cancers in the world with a high recurrence rate. In recent years, different researches have focused on designing efficient multi-epitope peptide vaccines against HCC. In designing these vaccines, over-expressed antigens in HCC patients, such as α-fetoprotein (AFP) and glypican-3 (GPC-3), have been employed. In our previous study, a multi-epitope peptide vaccine for HCC was designed by in-silico methods. The designed vaccine construct included the AFP, GPC-3, and aspartyl-β-hydroxylase (ASPH) as CytoLoxic T cell Lymphocytes (CTL), one epitope from Tetanus Toxin Fragment C (TTFrC) as Helper T cell Lymphocytes (HTL), and a segment of microbial heat shock protein (HSP70) peptide407-426 as an adjuvant. All the mentioned parts were connected by appropriate linkers. The aim of this study is the production of the designed vaccine. Methods: This research is experimental and was carried out in Fasa, Iran, in 2017. The designed vaccine construct gene was transformed to the Escherchia coli BL21 (DE3) strain and expressed in different isopropyl β-D-1-thiogalactopyranoside (IPTG) concentrations (0.6 and 1 mM), times (4, 6, 8, 16 hours), and temperatures (25 and 37 °C). Then, the expressed protein was analyzed by Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and the Western blot methods. Results: The best conditions for protein expression were obtained in the Super Optimal Broth (SOB) medium at 37 °C after the induction of expression by 1 mM IPTG for six hour. Conclusion: The recombinant HCC vaccine was produced with a proper concentration. [ABSTRACT FROM AUTHOR]

Leishmaniasis is one of the major health problems in most countries of the world. Millions of people around the world are at risk for the disease. Given the prevalence of this parasite in Iran and developing countries and the emergence of resistance in some cases to existing drugs, developing an effective vaccine against leishmaniasis is necessary. This research aims to design a multi-epitope vaccine derived from LACK, LeIF, GP63, and SMT antigens of Leishmania major based on the combination of bioinformatics methods. The synthesized construct with 16.86 KDa was cloned and sub-cloned in pEGFP- N1 and pLEXSY-neo2, respectively. They were then transfected in promastigotes of L. tarentolae. After confirmation of expression, immunization was carried out in 8 groups of BALB/c mice (9 mice per group) three times at two-week intervals. Cellular immune responses were assessed before and after the challenge by L. major. Furthermore, at 3rd week post-infection, the survival rate, mean lesion size, and parasite burden were assessed. All vaccinated mice demonstrated partial immunity to higher IFN-γ levels than the control groups (P<0.05). Immunized mice with cytosolic complex (G1) indicated the highest levels of IFN-ɤ and ratio of IFN ɤ/ IL-4, the lowest levels of IL-4 and IL-10 compared to control and the other groups (P≤0.05), and produced a partial Th1 immune response. Mean lesion size and parasite burden of G1 and G5 reduced significantly compared to the other and control groups post-challenge (P<0.05). The outputs of our result could be a hopeful sign in the achievement of practical approaches as multi-epitope vaccines against Leishmania major. [ABSTRACT FROM AUTHOR]