Abstract
Purpose : To evaluate the immunogenicity and types of immune response of a quality-controlled modified recombinant hepatitis B surface antigen (HBsAg) plasmid encoding HBsAg in mice. Methods : The characterized plasmid DNA was used in the immunization of Balb/c mice. Three groups of mice were intramuscularly injected with three different concentrations (50, 25 and 10 μg/100 μL) of the modified plasmid. Humoral immune response was monitored by enzyme-linked immunosorbent assay (ELISA), while cellular immune response was investigated by analysis of spleen cytokine profile (TNFα, IFN γ and IL2) as well as CD69 expression level in CD4 and CD8 positive cells. Results : In general, the activated CD4 cells showing intracellular cytokines were higher than CD8 positive population of cells (p < 0.05). These findings indicate that the vaccine induced both a humoral and cellular immunity. Cytokine profile also showed high levels of TNFα, IFN γ and IL2 and CD69 expression in the group of animals immunized at a dose of 10 μg when compared to control group (p < 0.05). Conclusion : A 10 μg dose intramuscular injection of the modified DNA-based vaccine encoding HBsAg in mice induces both high humoral and cellular immune responses. Keywords : Hepatitis B virus, Plasmid DNA, Vaccine, Spleen cytokines, Humoral and cellular immune responses
Highlights
Hepatitis B (HB) is a potentially life-threatening liver infection caused by Hepatitis B virus (HBV) [1]
The gWizHBsAg plasmid was used for immunization of 21 female Balb/c mice aged 6–8 weeks bred in King Saud University (KSU) animal house and kept under specific pathogen free conditions
The humoral immunity was evaluated by detecting specific hepatitis B surface antigen (HBsAg) IgG and IgM induced by modified DNA HBsAg plasmids in BALB/c immunized mice
Summary
Hepatitis B (HB) is a potentially life-threatening liver infection caused by Hepatitis B virus (HBV) [1]. The purpose of the present investigation was to assess the immunogenicity of previously reported DNA-HBsAg - based vaccine comprising modified gWiz-HBsAg plasmid, characterized and quality-controlled by SalemBekhit et al [5], and type of immune responses produced in mice. The gWizHBsAg plasmid was used for immunization of 21 female Balb/c mice aged 6–8 weeks bred in King Saud University (KSU) animal house and kept under specific pathogen free conditions. These mice were divided into three groups (A, B and C), each group comprised seven mice. HBsAg-specific CTL assay was performed using panels of monoclonal antibodies used for the detection of surface markers and intracellular cytokines of immunized splenocytes. Statistical significance of differences was defined as p < 0.05
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