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Immune response induced by recombinant pres2/S-protein and a pres2-S-protein fused with a core 18-27 antigen fragment of hepatitis B virus compared to conventional HBV vaccine

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Abstract

Although comprehensive vaccination is the cornerstone of public health programs to control hepatitis B virus (HBV) infections, 5% of people who receive the existing vaccine do not develop proper immunity against HBV. To overcome this challenge, researchers have tried using various protein fragments encoded by the virus genome to achieve better immunization rates. An important antigenic component of HBsAg called the preS2/S or M protein has also received much attention in this area. The gene sequences of preS2/S and Core18-27 peptide were extracted from the GenBank (NCBI). Final gene synthesis was conducted with pET28. Groups of BALB/c mice were immunized with 10 μg/ml of recombinant proteins and 1 μg/ml CPG7909 adjuvant. Serum levels of IF-γ, TNF-α, IL-2, IL-4, and IL-10 were measured by ELISA assay method on spleen cell cultures on day 45, and IgG1, IgG2a, and total IgG titers obtained from mice serum were quantified on days 14 and 45. Statistical analysis did not show any significant difference between the groups regarding IF-γ level. There were, however, significant differences in terms of IL-2 and IL-4 levels between the groups receiving preS2/S-C18-27 with and without adjuvant and the groups receiving both preS2/S and preS2/S-C18-27 (Plus Recomb—Plus Recomb: the group of mice that received both preS2/S and preS2/S-C18-27 simultaneously). The strongest total antibody production was induced by immunization with both recombinant proteins without CPG adjuvant. The groups that received both preS2/S and preS2/S-C18-27, whether with or without adjuvant, were significantly different from those that received the conventional vaccine considering most abundant interleukins. This difference suggested that higher levels of efficacy can be achieved by the use of multiple virus antigen fragments rather than using a single fragment.

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Data availability

The datasets used and/or analyzed during this study are available from the corresponding authors upon reasonable request.

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Acknowledgements

The authors are grateful to the staff of Clinical Microbiology Center, Ilam University of Medical Sciences, Ilam, Iran and the Applied Microbiology Research Center of Baqiyatallah University, Tehran, Iran.

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Authors and Affiliations

  1. Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran

    Elaheh Gholami Parizad & Afra Khosravi

  2. Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Vanak Sq. Mollasadra St., P.O. Box 19395-5487, Tehran, Iran

    Abbas Ali Imani Fooladi, Hamid Sedighian & Jafar Amani

  3. Academy of Medical Sciences of the I.R. of Iran, Tehran, Iran

    Elham Behzadi

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  1. Elaheh Gholami Parizad
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Contributions

EP, AI, and AK conceived the study and designed experiments. EP, HS, and JA performed experiments. EP, HS, and AI analyzed experiments. EP, EB, HS, AI, and AK wrote the manuscript with support from all authors. All authors read and approved the final manuscript.

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Correspondence to Abbas Ali Imani Fooladi or Afra Khosravi.

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The authors declare no conflict of interest associated with the present manuscript.

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The experimental protocol was reviewed, approved, and supervised by the institution of animal care and the scientific committee of Ilam University of Medical Sciences (permit number: IR.MEDILAM.REC.1395.105).

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During the experiment, the vaccinated mice were monitored every day. All surgeries were performed under sodium pentobarbital anesthesia and all efforts were made to minimize the suffering of animals. The mice were sacrificed by using cervical dislocation method.

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Parizad, E.G., Imani Fooladi, A.A., Sedighian, H. et al. Immune response induced by recombinant pres2/S-protein and a pres2-S-protein fused with a core 18-27 antigen fragment of hepatitis B virus compared to conventional HBV vaccine. Virus Genes 59, 499–514 (2023). https://doi.org/10.1007/s11262-023-01995-z

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