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<p>The emergence of immune-evasive variants of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) demonstrates the limitations of first-generation vaccines and underscores the necessity for broadly protective, next-generation vaccines. This study provides a proof-of-concept for a novel vaccine strategy based on the rational design and immunogenic evaluation of Johnsongrass mosaic virus-derived potyvirus-like particles (PVLPs) engineered to display conserved SARS-CoV-2 spike protein epitopes. Three conserved epitopes of the S2 subunit, VQ (991–1005 aa), TQ (1009–1023 aa), and MQ (896–910 aa) were identified through bioinformatic analysis and genetically fused to the N-terminus of the Johnsongrass mosaic virus coat protein. The resulting chimeric PVLPs were expressed and purified from <i>Escherichia coli</i> BL21(DE3) and characterized using SDS-PAGE. High-resolution transmission electron microscopy confirmed the successful assembly of the chimeric PVLPs. Humoral and cell-mediated immune responses to the chimeric PVLPs were assessed in pre-clinical studies. In mouse immunization experiments, the TQ: CP PVLP construct generated potent antibody titers following a single booster dose. The platform also induced a multifaceted cellular immune response, with epitope-dependent polarization of T-helper profiles. Specifically, the VQ: CP and MQ: CP PVLPs elicited a robust Th1-skewed response, characterized by high IFN-γ and low IL-4 levels. A mixed PVLP formulation produced a balanced Th1/Th2 response, demonstrating the capacity to tailor immune polarization through rational epitope selection. Immunization studies in mice showed that the chimeric PVLPs induced stronger humoral and cell-mediated immune responses compared to wild-type PVLP. These results establish the JGMV PVLP platform as a promising approach for developing a pan-coronavirus vaccine that displays conserved SARS-CoV-2 epitopes.</p>

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Chimeric potyvirus-like particle displaying conserved SARS-CoV-2 spike epitopes

  • 期刊论文

  • 发表日期：

    2026年3月17日

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  • A. Annie Aglin,
  • M.L Stephen Raj

摘要

The emergence of immune-evasive variants of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) demonstrates the limitations of first-generation vaccines and underscores the necessity for broadly protective, next-generation vaccines. This study provides a proof-of-concept for a novel vaccine strategy based on the rational design and immunogenic evaluation of Johnsongrass mosaic virus-derived potyvirus-like particles (PVLPs) engineered to display conserved SARS-CoV-2 spike protein epitopes. Three conserved epitopes of the S2 subunit, VQ (991–1005 aa), TQ (1009–1023 aa), and MQ (896–910 aa) were identified through bioinformatic analysis and genetically fused to the N-terminus of the Johnsongrass mosaic virus coat protein. The resulting chimeric PVLPs were expressed and purified from Escherichia coli BL21(DE3) and characterized using SDS-PAGE. High-resolution transmission electron microscopy confirmed the successful assembly of the chimeric PVLPs. Humoral and cell-mediated immune responses to the chimeric PVLPs were assessed in pre-clinical studies. In mouse immunization experiments, the TQ: CP PVLP construct generated potent antibody titers following a single booster dose. The platform also induced a multifaceted cellular immune response, with epitope-dependent polarization of T-helper profiles. Specifically, the VQ: CP and MQ: CP PVLPs elicited a robust Th1-skewed response, characterized by high IFN-γ and low IL-4 levels. A mixed PVLP formulation produced a balanced Th1/Th2 response, demonstrating the capacity to tailor immune polarization through rational epitope selection. Immunization studies in mice showed that the chimeric PVLPs induced stronger humoral and cell-mediated immune responses compared to wild-type PVLP. These results establish the JGMV PVLP platform as a promising approach for developing a pan-coronavirus vaccine that displays conserved SARS-CoV-2 epitopes.