Antibodies against VP3 protein of echovirus 30 (<i>Picornaviridae: Enterovirus: Enterovirus betacoxsackie</i>) neutralize virus <i>in vitro</i>
2025
Dmitry V. Novikov | Dmitry A. Melentev | Vladimir Yu. Talayev | Nadezhda A. Novikova | Ekaterina V. Mokhonova | Alexander U. Kashnikov | Irina E. Zaichenko | Marya V. Svetlova | Elena V. Kurkova | Olga N. Babaykina | Vladislav A. Lapin | Maria I. Tsyganova | Dmitry E. Zaitsev | Viktor V. Novikov
Introduction. The widespread occurrence of enterovirus echovirus 30 (E30) and cases of severe disease indicate the need for vaccine development. Epitopes for neutralizing antibodies and T-cell response have been found in VP3 proteins of some enteroviruses. However, the immunogenic properties of VP3 E30 have not been studied. The aim of this work is to characterize the immunogenic properties of VP3 E30 and testing the virus-neutralizing properties of antibodies against VP3 E30. Materials and methods. VP3E30 and the chimeric protein SN-VP3E30, consisting of the S region of norovirus VP1 and VP3 E30 were expressed in Escherichia coli. SN-VP3E30 was used to form virus-like particles (VLPs). The effect on human dendritic cells (DCs) was assessed by flow cytometry to measure changes in HLA-DR, CCR7, CD80, CD83 and CD86 expression. BALB/c mice and guinea pig were used for immunization with SN-VP3E30. Antibody titers and avidity were determined by ELISA. The interaction of antibodies with VP3 E30 was studied by immunoelectron microscopy and virus neutralization in embryonal rhabdomyosarcoma (RD) cell culture. Results. Recombinant VP3 E30 caused incomplete DCs maturation, characterized by the understimulation of the CCR7 chemokine receptor expression. Inclusion of VP3 in the chimeric VLPs resulted in complete DCs maturation and a strong humoral immune response in laboratory animals. Antibodies against VP3 were characterized by high avidity, the ability to induce agglomeration of viral particles and neutralization of E30 in RD cell culture. Conclusion. The obtained results indicate that VP3 can be used as an antigen in the composition of a subunit vaccine against enterovirus E30.
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