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KMID : 1039120230120010047
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Clinical and Experimental Vaccine Research
2023 Volume.12 No. 1 p.47 ~ p.59
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Individual expression and processing of hepatitis C virus E1/E2 epitopes-based DNA vaccine candidate in healthy humans’ peripheral blood mononuclear cells
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Rola Nadeem
Amany Sayed Maghraby
Dina Nadeem Abd-Elshafy
Ahmed Barakat Barakat
Mahmoud Mohamed Bahgat
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Abstract
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Purpose: The development and study of hepatitis C virus (HCV) vaccine candidates’ individualized responses are of great importance. Here we report on an HCV DNA vaccine candidate based on selected envelope (E1/E2) epitopes. Besides, we assessed its expression and processing in human peripheral blood mononuclear cells (PBMCs) and in vivo cellular response in mice.
Materials and Methods: HCV E1/E2 DNA construct (EC) was designed. The antigen expression of EC was assayed in PBMCs of five HCV-uninfected donors via a real-time quantitative polymerase chain reaction. Serum samples from 20 HCV antibody-positive patients were used to detect each individual PBMCs expressed antigens via enzyme-linked immunosorbent assay. Two groups, five Swiss albino mice each, were immunized with the EC or a control construct. The absolute count of lymph nodes’ CD4+ and CD8+ T-lymphocytes was assessed.
Results: Donors’ PBMCs showed different levels of EC expression, ranging between 0.83?2.61-fold in four donors, while donor-3 showed 34.53-fold expression. The antigens expressed in PBMCs were significantly reactive to the 20 HCV antibody repertoire (all p=0.0001). All showed comparable reactivity except for donor-3 showing the lowest reactivity level. The absolute count % of the CD4+ T-cell significantly increased in four of the five EC-immunized mice compared to the control group (p=0.03). No significant difference in CD8+ T-cells % was observed (p=0.89).
Conclusion: The inter-individual variation in antigen expression and processing dominance was evident, showing independence in individuals’ antigen expression and reactivity levels to antibodies. The described vaccine candidate might result in a promising natural immune response with a possibility of CD4+ T-cell early priming.
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KEYWORD
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Hepatitis C virus, DNA vaccines, Human peripheral blood mononuclear cells, Individuality, Envelope protein
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