DESIGN OF POLYVALENT HEPATITIS VACCINE CANDIDATE AND EVALUATION OF ITS EX VIVO STIMULATION IN HUMAN DENDRITIC CELLS

Background and Aim:Hepatitis C Virus (HCV) the main cause of chronic hepatitis; Annually, about 185 million people are infected with HCV and despite many efforts, it remains a serious burden to global public health. Thus, vaccine design based on the new approach is an appropriate alternative for this problem. In this study, besides the immunodominant epitopes of HCV core, in order to enhance the efficiency of designed polyvalent vaccine and induce a robust immune response, Hepatitis B surface antigen (HBsAg) and polio virus VP antigen were used as antigenic determinants.Methods:Epitopes identification carried out for each antigen using various immunoinformatics tools and analysis. The selected epitopes were fused together by proper linkers. The entire features of final protein were evaluated by different servers and downstream analyses were performed subsequently. Polyepitope vaccine construct was chemically synthesized in pET28a expression vector. Protein expression in E.Coli was induced by adding IPTG 1mM. Purification was performed using affinity chromatography. In the next step, the immune stimulation of the desired vaccine evaluated in dendritic cells derived from human peripheral blood mononuclear cells and its effect on the activation and proliferation of T cells and cytokines secretion were assessed by flow cytometry and Elisa.Results:The results indicated that the designed polyepitope has better or in some cases at least equal effect in comparison to the monovalent vaccine to induce T-cell activation and proliferation.Conclusion:Our designed vaccine candidate has a capacity for simultaneous immunization against three viral diseases and induction of proper immune response.

کلیدواژه ها:

Hepatitis C Virus ، PolyEpitope ، Polyvalent Vaccine ، Dendritic Cells ، Flow Cytometry