IMMUNOINFORMATICS MODELING, CONSTRUCTION OF DNA PLASMIDS CARRYING CTL EPITOPES OF HEPATITIS C VIRUS AND THEIR PRELIMINARY IMMUNOLOGICAL ANALYSIS

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Journal: IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY Year:2011 | Volume:4 | Issue:4 Page(s): 30-40

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Title

IMMUNOINFORMATICS MODELING, CONSTRUCTION OF DNA PLASMIDS CARRYING CTL EPITOPES OF HEPATITIS C VIRUS AND THEIR PRELIMINARY IMMUNOLOGICAL ANALYSIS

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, , , , , | Issue Writer Certificate

Keywords

HEPATITIS C VIRUSQ4

IMMUNOINFONNATICSQ4

MULTIEPITOPE VACCINEQ4

IN VIVO EXPRESSIONQ4

IMMUNOSTIMULANTQ4

Abstract

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, El 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, Westem- 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.

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APA: Copy

. (2011). IMMUNOINFORMATICS MODELING, CONSTRUCTION OF DNA PLASMIDS CARRYING CTL EPITOPES OF HEPATITIS C VIRUS AND THEIR PRELIMINARY IMMUNOLOGICAL ANALYSIS. IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY, 4(4), 30-40. SID. https://sid.ir/paper/132733/en

Vancouver: Copy

. IMMUNOINFORMATICS MODELING, CONSTRUCTION OF DNA PLASMIDS CARRYING CTL EPITOPES OF HEPATITIS C VIRUS AND THEIR PRELIMINARY IMMUNOLOGICAL ANALYSIS. IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY[Internet]. 2011;4(4):30-40. Available from: https://sid.ir/paper/132733/en

IEEE: Copy

, “IMMUNOINFORMATICS MODELING, CONSTRUCTION OF DNA PLASMIDS CARRYING CTL EPITOPES OF HEPATITIS C VIRUS AND THEIR PRELIMINARY IMMUNOLOGICAL ANALYSIS,” IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY, vol. 4, no. 4, pp. 30–40, 2011, [Online]. Available: https://sid.ir/paper/132733/en

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