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Background <p>Akabane virus (AKAV) is the causative agent of an economically significant disease in ruminants, manifested notably by outbreaks of abortion and congenital abnormalities. Vaccination stands as the primary defense against this disease. However, the development of safer, more stable, and efficient AKAV vaccines, including epitope-based designs, remains unexplored. Prior work by our group has pinpointed a neutralizing epitope, <sup>1134</sup>SVQSFDGKL<sup>1142</sup>, located in the Gc protein of AKAV. We further demonstrated its high degree of conservation across diverse AKAV genotypes.</p> Methods <p>&#xa0;We produced and verified a novel virus-like particle (VLP) by incorporating the neutralizing epitope <sup>1134</sup>SVQSFDGKL<sup>1142</sup> into a recombinant hepatitis B virus core antigen (HBcAg) scaffold. Then the immunogenicity of this VLP was evaluated by detecting the antibody titer targeting the AKAV Gc antigen and the neutralizing activity against AKAV in sera from the VLP-immunized mice. Furthermore, a preliminary indirect ELISA method was established based on this VLP for AKAV detection.</p> Results <p>The successful construction of VLP expressing the AKAV epitope was confirmed by using SDS-PAGE, followed by Western blot (WB) and transmission electron microscopy (TEM). Indirect ELISA results indicated that antisera from immunized mice contained antibodies specific to the AKAV Gc protein. Furthermore, neutralization assays demonstrated that the antisera could effectively neutralize AKAV in vitro and inhibit its replication in BHK-21 cells. The developed VLP-based indirect ELISA method successfully identified AKAV antibody-positive serum, with a detection sensitivity of up to a 1:1600 serum dilution.</p> Conclusions <p>In conclusion, we successfully constructed a VLP presenting the highly conserved neutralizing epitope of AKAV. This VLP is proved to be immunogenic and can serve as an effective coating antigen to establish an indirect ELISA method for AKAV detection. Collectively, our findings provide proof-of-concept for this epitope-presenting VLP as a promising candidate in the pursuit of a safe and effective epitope-based vaccine against AKAV and also highlight its utility as a diagnostic antigen for serological detection.</p>

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Immunogenicity analyses and indirect ELISA application of a chimeric virus-like particle presenting a highly conserved peptide of Akabane virus Gc protein

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月30日

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  • Jingjing Wang,
  • Fang Wei,
  • Ruyang Yu,
  • Dongjie Chen,
  • Shaoqiang Wu

摘要

Background

Akabane virus (AKAV) is the causative agent of an economically significant disease in ruminants, manifested notably by outbreaks of abortion and congenital abnormalities. Vaccination stands as the primary defense against this disease. However, the development of safer, more stable, and efficient AKAV vaccines, including epitope-based designs, remains unexplored. Prior work by our group has pinpointed a neutralizing epitope, 1134SVQSFDGKL1142, located in the Gc protein of AKAV. We further demonstrated its high degree of conservation across diverse AKAV genotypes.

Methods

 We produced and verified a novel virus-like particle (VLP) by incorporating the neutralizing epitope 1134SVQSFDGKL1142 into a recombinant hepatitis B virus core antigen (HBcAg) scaffold. Then the immunogenicity of this VLP was evaluated by detecting the antibody titer targeting the AKAV Gc antigen and the neutralizing activity against AKAV in sera from the VLP-immunized mice. Furthermore, a preliminary indirect ELISA method was established based on this VLP for AKAV detection.

Results

The successful construction of VLP expressing the AKAV epitope was confirmed by using SDS-PAGE, followed by Western blot (WB) and transmission electron microscopy (TEM). Indirect ELISA results indicated that antisera from immunized mice contained antibodies specific to the AKAV Gc protein. Furthermore, neutralization assays demonstrated that the antisera could effectively neutralize AKAV in vitro and inhibit its replication in BHK-21 cells. The developed VLP-based indirect ELISA method successfully identified AKAV antibody-positive serum, with a detection sensitivity of up to a 1:1600 serum dilution.

Conclusions

In conclusion, we successfully constructed a VLP presenting the highly conserved neutralizing epitope of AKAV. This VLP is proved to be immunogenic and can serve as an effective coating antigen to establish an indirect ELISA method for AKAV detection. Collectively, our findings provide proof-of-concept for this epitope-presenting VLP as a promising candidate in the pursuit of a safe and effective epitope-based vaccine against AKAV and also highlight its utility as a diagnostic antigen for serological detection.