AU - Arashkia, Arash AU - Rouhvand, Farzin AU - Memarnejadian, Arash AU - Alizadeh, Shahadeh AU - Motevalli, Fatemeh AU - Ebrahimi, Mekania TI - Immunoinformatics modeling, construction of DNA plasmids Carrying CTL epitopes of hepatitis C virus and their preliminary immunological analysis PT - JOURNAL ARTICLE TA - Iran-J-Med-Microbiol JN - Iran-J-Med-Microbiol VO - 4 VI - 4 IP - 4 4099 - http://ijmm.ir/article-1-25-en.html 4100 - http://ijmm.ir/article-1-25-en.pdf SO - Iran-J-Med-Microbiol 4 AB  - Background and objectives: High mutation rate and immunosuppressive effect of some key proteins of hepatitis C virus (HCV) encouraged researchers to find conserved and small immunogenic units to induce immune responses. Thus epitope-based vaccines were introduced to find out any potential immunogens. However, the relatively weak immune responses against these vaccines require a creation of additional measures. This study aimed to construct plasmids carrying CTL epitope of hepatitis C virus using immunoinformatics modeling method and assessment of their preliminary immunogenicity.Materials and Methods: One H-2Dd and two HLA-A*0201-restricted CD8+ T-cell epitopes from E2, E1 and core regions of HCV were selected and immunoinformatically analyzed for optimum sequentiality. The multiepitope sequence was constructed by SOEing PCR and cloned in pcDNA3.1+ vector. PADRE, endoplasmic reticulum signal sequence (ERss) and hepatitis B surface antigen (HBsAg) immuno-enhancer sequences were fused to the 5′ end of multiepitope constructs. In vitro, expression of each plasmid was analyzed by RT-PCR, dot-blot, Western-blot and immunofluorescence techniques. Preliminary in vivo immunogenicity of the construct was assessed by delayed-type hypersensitivity (DTH) response in BALB/c mice.Results: In vitro, analyses confirmed the expression of plasmids. Preliminary, in vivo assessments indicated the processing and presentation of the H-2Dd epitope in BALB/c mice. Moreover, although the immunostimulation effects of ERss and PADRE were shown, fusion of HBsAg did not enhance the DTH response. Conclusion: This study shows the value of immunoinformatics prediction of hepatitis C virus epitopes as a multiepitope vaccine candidate and DTH assay for preliminary analysis using BALB/c mice. Data obtained, provide enough support for further evaluation of the designed constructs in HLA-A2 transgenic mice. CP - IRAN IN - LG - eng PB - Iran-J-Med-Microbiol PG - 30 PT - Original Research Article YR - 2011